Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells

<!DOCTYPE html> <html lang="en" xmlns:og="http://ogp.me/ns#" xmlns:fb="https://www.facebook.com/2008/fbml"> <head> <meta charset="utf-8"> <meta http-equiv="X-UA-Compatible" content="IE=edge,chrome=1"> <meta content="mdpi" name="sso-service" /> <meta content="width=device-width, initial-scale=1.0" name="viewport" /> <title>Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells</title><link rel="stylesheet" href="https://pub.mdpi-res.com/assets/css/font-awesome.min.css?eb190a3a77e5e1ee?1732615622"> <link rel="stylesheet" href="https://pub.mdpi-res.com/assets/css/jquery.multiselect.css?f56c135cbf4d1483?1732615622"> <link rel="stylesheet" href="https://pub.mdpi-res.com/assets/css/chosen.min.css?d7ca5ca9441ef9e1?1732615622"> <link rel="stylesheet" href="https://pub.mdpi-res.com/assets/css/main2.css?69b39374e6b554b7?1732615622"> <link rel="mask-icon" href="https://pub.mdpi-res.com/img/mask-icon-128.svg?c1c7eca266cd7013?1732615622" color="#4f5671"> <link rel="apple-touch-icon" sizes="180x180" href="https://pub.mdpi-res.com/icon/apple-touch-icon-180x180.png?1732615622"> <link rel="apple-touch-icon" sizes="152x152" href="https://pub.mdpi-res.com/icon/apple-touch-icon-152x152.png?1732615622"> <link rel="apple-touch-icon" sizes="144x144" href="https://pub.mdpi-res.com/icon/apple-touch-icon-144x144.png?1732615622"> <link rel="apple-touch-icon" sizes="120x120" href="https://pub.mdpi-res.com/icon/apple-touch-icon-120x120.png?1732615622"> <link rel="apple-touch-icon" sizes="114x114" href="https://pub.mdpi-res.com/icon/apple-touch-icon-114x114.png?1732615622"> <link rel="apple-touch-icon" sizes="76x76" href="https://pub.mdpi-res.com/icon/apple-touch-icon-76x76.png?1732615622"> <link rel="apple-touch-icon" sizes="72x72" href="https://pub.mdpi-res.com/icon/apple-touch-icon-72x72.png?1732615622"> <link rel="apple-touch-icon" sizes="57x57" href="https://pub.mdpi-res.com/icon/apple-touch-icon-57x57.png?1732615622"> <link rel="apple-touch-icon" href="https://pub.mdpi-res.com/icon/apple-touch-icon-57x57.png?1732615622"> <link rel="apple-touch-icon-precomposed" href="https://pub.mdpi-res.com/icon/apple-touch-icon-57x57.png?1732615622"> <link rel="manifest" href="/manifest.json"> <meta name="theme-color" content="#ffffff"> <meta name="application-name" content=" "/> <link rel="apple-touch-startup-image" href="https://pub.mdpi-res.com/img/journals/biomedicines-logo-sq.png?8600e93ff98dbf14"> <link rel="apple-touch-icon" href="https://pub.mdpi-res.com/img/journals/biomedicines-logo-sq.png?8600e93ff98dbf14"> <meta name="msapplication-TileImage" content="https://pub.mdpi-res.com/img/journals/biomedicines-logo-sq.png?8600e93ff98dbf14"> <link rel="stylesheet" href="https://pub.mdpi-res.com/assets/css/jquery-ui-1.10.4.custom.min.css?80647d88647bf347?1732615622"> <link rel="stylesheet" href="https://pub.mdpi-res.com/assets/css/magnific-popup.min.css?04d343e036f8eecd?1732615622"> <link rel="stylesheet" href="https://pub.mdpi-res.com/assets/css/xml2html/article-html.css?230b005b39af4260?1732615622"> <style> h2, #abstract .related_suggestion_title { } .batch_articles a { color: #000; } a, .batch_articles .authors a, a:focus, a:hover, a:active, .batch_articles a:focus, .batch_articles a:hover, li.side-menu-li a { } span.label a { color: #fff; } #main-content a.title-link:hover, #main-content a.title-link:focus, #main-content div.generic-item a.title-link:hover, #main-content div.generic-item a.title-link:focus { } #main-content #middle-column .generic-item.article-item a.title-link:hover, #main-content #middle-column .generic-item.article-item a.title-link:focus { } .art-authors a.toEncode { color: #333; font-weight: 700; } #main-content #middle-column ul li::before { } .accordion-navigation.active a.accordion__title, .accordion-navigation.active a.accordion__title::after { } .accordion-navigation li:hover::before, .accordion-navigation li:hover a, .accordion-navigation li:focus a { } .relative-size-container .relative-size-image .relative-size { } .middle-column__help__fixed a:hover i, } input[type="checkbox"]:checked:after { } input[type="checkbox"]:not(:disabled):hover:before { } #main-content .bolded-text { } #main-content .hypothesis-count-container { } #main-content .hypothesis-count-container:before { } .full-size-menu ul li.menu-item .dropdown-wrapper { } .full-size-menu ul li.menu-item > a.open::after { } #title-story .title-story-orbit .orbit-caption { #background: url('/img/design/000000_background.png') !important; background: url('/img/design/ffffff_background.png') !important; color: rgb(51, 51, 51) !important; } #main-content .content__container__orbit { background-color: #000 !important; } #main-content .content__container__journal { color: #fff; } .html-article-menu .row span { } .html-article-menu .row span.active { } .accordion-navigation__journal .side-menu-li.active::before, .accordion-navigation__journal .side-menu-li.active a { color: rgba(20,64,153,0.75) !important; font-weight: 700; } .accordion-navigation__journal .side-menu-li:hover::before , .accordion-navigation__journal .side-menu-li:hover a { color: rgba(20,64,153,0.75) !important; } .side-menu-ul li.active a, .side-menu-ul li.active, .side-menu-ul li.active::before { color: rgba(20,64,153,0.75) !important; } .side-menu-ul li.active a { } .result-selected, .active-result.highlighted, .active-result:hover, .result-selected, .active-result.highlighted, .active-result:focus { } .search-container.search-container__default-scheme { } nav.tab-bar .open-small-search.active:after { } .search-container.search-container__default-scheme .custom-accordion-for-small-screen-link::after { color: #fff; } @media only screen and (max-width: 50em) { #main-content .content__container.journal-info { color: #fff; } #main-content .content__container.journal-info a { color: #fff; } } .button.button--color { } .button.button--color:hover, .button.button--color:focus { } .button.button--color-journal { position: relative; background-color: rgba(20,64,153,0.75); border-color: #fff; color: #fff !important; } .button.button--color-journal:hover::before { content: ''; position: absolute; top: 0; left: 0; height: 100%; width: 100%; background-color: #ffffff; opacity: 0.2; } .button.button--color-journal:visited, .button.button--color-journal:hover, .button.button--color-journal:focus { background-color: rgba(20,64,153,0.75); border-color: #fff; color: #fff !important; } .button.button--color path { } .button.button--color:hover path { fill: #fff; } #main-content #search-refinements .ui-slider-horizontal .ui-slider-range { } .breadcrumb__element:last-of-type a { } #main-header { } #full-size-menu .top-bar, #full-size-menu li.menu-item span.user-email { } .top-bar-section li:not(.has-form) a:not(.button) { } #full-size-menu li.menu-item .dropdown-wrapper li a:hover { } #full-size-menu li.menu-item a:hover, #full-size-menu li.menu.item a:focus, nav.tab-bar a:hover { } #full-size-menu li.menu.item a:active, #full-size-menu li.menu.item a.active { } #full-size-menu li.menu-item a.open-mega-menu.active, #full-size-menu li.menu-item div.mega-menu, a.open-mega-menu.active { } #full-size-menu li.menu-item div.mega-menu li, #full-size-menu li.menu-item div.mega-menu a { border-color: #9a9a9a; } div.type-section h2 { font-size: 20px; line-height: 26px; font-weight: 300; } div.type-section h3 { margin-left: 15px; margin-bottom: 0px; font-weight: 300; } .journal-tabs .tab-title.active a { } </style> <link rel="stylesheet" href="https://pub.mdpi-res.com/assets/css/slick.css?f38b2db10e01b157?1732615622"> <meta name="title" content="Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells"> <meta name="description" content="Glioblastoma (GBM) is the most aggressive cancer originating in the brain, but unfortunately combination treatments with resection, radiation, and chemotherapy are relatively ineffective. Therefore, novel methods of adjuvant therapy are critically needed. Cyclotides are plant-derived circular peptides that chemosensitize drug-resistant breast cancer to doxorubicin. We analyzed naturally occurring and synthetic cyclotides (Cycloviolacin O3, Cycloviolacin O19, natural Kalata B1, synthetic Kalata B1, and Vitri E) alone and in co-exposure treatments with the drug temozolomide (TMZ) in human glioblastoma cells. The cyclotides were identified by UPLC-PDA and HPLC-UV. The synthetic Kalata B1 sequence was verified with orbitrap LC-MS, and structural confirmation was provided by NMR spectroscopy. The cyclotides displayed dose-dependent cytotoxicity (IC50 values 2.4–21.1 µM) both alone and as chemosensitizers of U-87 MG and T 98 cells to TMZ. In fact, a 16-fold lower concentration of TMZ (100 µM) was needed for significant cytotoxicity in U-87 MG cells co-exposed to synthetic Kalata B (0.5 µM). Similarly, a 15-fold lower concentration of TMZ (75 µM) was required for a significant reduction in cell viability in T 98 cells co-exposed to synthetic Kalata B1 (0.25 µM). Kalata B1 remained stable in human serum stability assays. The data support the assertion that cyclotides may chemosensitize glioblastoma cells to TMZ." > <link rel="image_src" href="https://pub.mdpi-res.com/img/journals/biomedicines-logo.png?8600e93ff98dbf14" > <meta name="dc.title" content="Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells"> <meta name="dc.creator" content="Samantha L. Gerlach"> <meta name="dc.creator" content="James S. Metcalf"> <meta name="dc.creator" content="Rachael A. Dunlop"> <meta name="dc.creator" content="Sandra Anne Banack"> <meta name="dc.creator" content="Cheenou Her"> <meta name="dc.creator" content="Viswanathan V. Krishnan"> <meta name="dc.creator" content="Ulf Göransson"> <meta name="dc.creator" content="Sunithi Gunasekera"> <meta name="dc.creator" content="Blazej Slazak"> <meta name="dc.creator" content="Paul Alan Cox"> <meta name="dc.type" content="Article"> <meta name="dc.source" content="Biomedicines 2024, Vol. 12, Page 2216"> <meta name="dc.date" content="2024-09-28"> <meta name ="dc.identifier" content="10.3390/biomedicines12102216"> <meta name="dc.publisher" content="Multidisciplinary Digital Publishing Institute"> <meta name="dc.rights" content="http://creativecommons.org/licenses/by/3.0/"> <meta name="dc.format" content="application/pdf" > <meta name="dc.language" content="en" > <meta name="dc.description" content="Glioblastoma (GBM) is the most aggressive cancer originating in the brain, but unfortunately combination treatments with resection, radiation, and chemotherapy are relatively ineffective. Therefore, novel methods of adjuvant therapy are critically needed. Cyclotides are plant-derived circular peptides that chemosensitize drug-resistant breast cancer to doxorubicin. We analyzed naturally occurring and synthetic cyclotides (Cycloviolacin O3, Cycloviolacin O19, natural Kalata B1, synthetic Kalata B1, and Vitri E) alone and in co-exposure treatments with the drug temozolomide (TMZ) in human glioblastoma cells. The cyclotides were identified by UPLC-PDA and HPLC-UV. The synthetic Kalata B1 sequence was verified with orbitrap LC-MS, and structural confirmation was provided by NMR spectroscopy. The cyclotides displayed dose-dependent cytotoxicity (IC50 values 2.4–21.1 µM) both alone and as chemosensitizers of U-87 MG and T 98 cells to TMZ. In fact, a 16-fold lower concentration of TMZ (100 µM) was needed for significant cytotoxicity in U-87 MG cells co-exposed to synthetic Kalata B (0.5 µM). Similarly, a 15-fold lower concentration of TMZ (75 µM) was required for a significant reduction in cell viability in T 98 cells co-exposed to synthetic Kalata B1 (0.25 µM). Kalata B1 remained stable in human serum stability assays. The data support the assertion that cyclotides may chemosensitize glioblastoma cells to TMZ." > <meta name="dc.subject" content="cyclotides" > <meta name="dc.subject" content="glioblastoma" > <meta name="dc.subject" content="temozolomide" > <meta name="dc.subject" content="chemosensitize" > <meta name="dc.subject" content="chemotherapy" > <meta name="dc.subject" content="adjuvant therapy" > <meta name="dc.subject" content="<i>Viola</i>" > <meta name ="prism.issn" content="2227-9059"> <meta name ="prism.publicationName" content="Biomedicines"> <meta name ="prism.publicationDate" content="2024-09-28"> <meta name ="prism.volume" content="12"> <meta name ="prism.number" content="10"> <meta name ="prism.section" content="Article" > <meta name ="prism.startingPage" content="2216" > <meta name="citation_issn" content="2227-9059"> <meta name="citation_journal_title" content="Biomedicines"> <meta name="citation_publisher" content="Multidisciplinary Digital Publishing Institute"> <meta name="citation_title" content="Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells"> <meta name="citation_publication_date" content="2024/10"> <meta name="citation_online_date" content="2024/09/28"> <meta name="citation_volume" content="12"> <meta name="citation_issue" content="10"> <meta name="citation_firstpage" content="2216"> <meta name="citation_author" content="Gerlach, Samantha L."> <meta name="citation_author" content="Metcalf, James S."> <meta name="citation_author" content="Dunlop, Rachael A."> <meta name="citation_author" content="Banack, Sandra Anne"> <meta name="citation_author" content="Her, Cheenou"> <meta name="citation_author" content="Krishnan, Viswanathan V."> <meta name="citation_author" content="Göransson, Ulf"> <meta name="citation_author" content="Gunasekera, Sunithi"> <meta name="citation_author" content="Slazak, Blazej"> <meta name="citation_author" content="Cox, Paul Alan"> <meta name="citation_doi" content="10.3390/biomedicines12102216"> <meta name="citation_id" content="mdpi-biomedicines12102216"> <meta name="citation_abstract_html_url" content="https://www.mdpi.com/2227-9059/12/10/2216"> <meta name="citation_pdf_url" content="https://www.mdpi.com/2227-9059/12/10/2216/pdf?version=1728463288"> <link rel="alternate" type="application/pdf" title="PDF Full-Text" href="https://www.mdpi.com/2227-9059/12/10/2216/pdf?version=1728463288"> <meta name="fulltext_pdf" content="https://www.mdpi.com/2227-9059/12/10/2216/pdf?version=1728463288"> <meta name="citation_fulltext_html_url" content="https://www.mdpi.com/2227-9059/12/10/2216/htm"> <link rel="alternate" type="text/html" title="HTML Full-Text" href="https://www.mdpi.com/2227-9059/12/10/2216/htm"> <meta name="fulltext_html" content="https://www.mdpi.com/2227-9059/12/10/2216/htm"> <link rel="alternate" type="text/xml" title="XML Full-Text" href="https://www.mdpi.com/2227-9059/12/10/2216/xml"> <meta name="fulltext_xml" content="https://www.mdpi.com/2227-9059/12/10/2216/xml"> <meta name="citation_xml_url" content="https://www.mdpi.com/2227-9059/12/10/2216/xml"> <meta name="twitter:card" content="summary" /> <meta name="twitter:site" content="@MDPIOpenAccess" /> <meta name="twitter:image" content="https://pub.mdpi-res.com/img/journals/biomedicines-logo-social.png?8600e93ff98dbf14" /> <meta property="fb:app_id" content="131189377574"/> <meta property="og:site_name" content="MDPI"/> <meta property="og:type" content="article"/> <meta property="og:url" content="https://www.mdpi.com/2227-9059/12/10/2216" /> <meta property="og:title" content="Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells" /> <meta property="og:description" content="Glioblastoma (GBM) is the most aggressive cancer originating in the brain, but unfortunately combination treatments with resection, radiation, and chemotherapy are relatively ineffective. Therefore, novel methods of adjuvant therapy are critically needed. Cyclotides are plant-derived circular peptides that chemosensitize drug-resistant breast cancer to doxorubicin. We analyzed naturally occurring and synthetic cyclotides (Cycloviolacin O3, Cycloviolacin O19, natural Kalata B1, synthetic Kalata B1, and Vitri E) alone and in co-exposure treatments with the drug temozolomide (TMZ) in human glioblastoma cells. The cyclotides were identified by UPLC-PDA and HPLC-UV. The synthetic Kalata B1 sequence was verified with orbitrap LC-MS, and structural confirmation was provided by NMR spectroscopy. The cyclotides displayed dose-dependent cytotoxicity (IC50 values 2.4–21.1 µM) both alone and as chemosensitizers of U-87 MG and T 98 cells to TMZ. In fact, a 16-fold lower concentration of TMZ (100 µM) was needed for significant cytotoxicity in U-87 MG cells co-exposed to synthetic Kalata B (0.5 µM). Similarly, a 15-fold lower concentration of TMZ (75 µM) was required for a significant reduction in cell viability in T 98 cells co-exposed to synthetic Kalata B1 (0.25 µM). Kalata B1 remained stable in human serum stability assays. The data support the assertion that cyclotides may chemosensitize glioblastoma cells to TMZ." /> <meta property="og:image" content="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g001-550.jpg?1728463370" /> <link rel="alternate" type="application/rss+xml" title="MDPI Publishing - Latest articles" href="https://www.mdpi.com/rss"> <meta name="google-site-verification" content="PxTlsg7z2S00aHroktQd57fxygEjMiNHydKn3txhvwY"> <meta name="facebook-domain-verification" content="mcoq8dtq6sb2hf7z29j8w515jjoof7" /> <script id="Cookiebot" data-cfasync="false" src="https://consent.cookiebot.com/uc.js" data-cbid="51491ddd-fe7a-4425-ab39-69c78c55829f" type="text/javascript" async></script>  <script type="text/plain" data-cookieconsent="statistics"> (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start': new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0], j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src= 'https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f); })(window,document,'script','dataLayer','GTM-WPK7SW5'); </script> <script type="text/plain" data-cookieconsent="statistics"> _linkedin_partner_id = "2846186"; window._linkedin_data_partner_ids = window._linkedin_data_partner_ids || []; window._linkedin_data_partner_ids.push(_linkedin_partner_id); </script><script type="text/javascript"> (function(){var s = document.getElementsByTagName("script")[0]; var b = document.createElement("script"); b.type = "text/javascript";b.async = true; b.src = "https://snap.licdn.com/li.lms-analytics/insight.min.js"; s.parentNode.insertBefore(b, s);})(); </script> <script type="text/plain" data-cookieconsent="statistics" data-cfasync="false" src="//script.crazyegg.com/pages/scripts/0116/4951.js" async="async" ></script> </head> <body> <div class="direction direction_right" id="small_right" style="border-right-width: 0px; padding:0;"> <i class="fa fa-caret-right fa-2x"></i> </div> <div class="big_direction direction_right" id="big_right" style="border-right-width: 0px;"> <div style="text-align: right;"> Next Article in Journal<br> <div><a href="/2227-9059/12/10/2217">Exploring Intra-Articular Administration of Monoclonal Antibodies as a Novel Approach to Osteoarthritis Treatment: A Systematic Review</a></div> </div> </div> <div class="direction" id="small_left" style="border-left-width: 0px"> <i class="fa fa-caret-left fa-2x"></i> </div> <div class="big_direction" id="big_left" style="border-left-width: 0px;"> <div> Previous Article in Journal<br> <div><a href="/2227-9059/12/10/2215">The Wound-Healing Effect of a Novel Fibroblasts-Impregnated Hydroxyethylcellulose Gel in a Rat Full-Thickness Burn Model: A Preclinical Study</a></div> </div> </div> <div style="clear: both;"></div> <div id="menuModal" class="reveal-modal reveal-modal-new reveal-modal-menu" aria-hidden="true" data-reveal role="dialog"> <div class="menu-container"> <div class="UI_NavMenu"> <div class="content__container " > <div class="custom-accordion-for-small-screen-link " > <h2>Journals</h2> </div> <div class="target-item custom-accordion-for-small-screen-content show-for-medium-up"> <div class="menu-container__links"> <div style="width: 100%; float: left;"> <a href="/about/journals">Active Journals</a> <a href="/about/journalfinder">Find a Journal</a> <a href="/about/journals/proposal">Journal Proposal</a> <a href="/about/proceedings">Proceedings Series</a> </div> </div> </div> </div> <a href="/topics"> <h2>Topics</h2> </a> <div class="content__container " > <div class="custom-accordion-for-small-screen-link " > <h2>Information</h2> </div> <div class="target-item custom-accordion-for-small-screen-content show-for-medium-up"> <div class="menu-container__links"> <div style="width: 100%; max-width: 200px; float: left;"> <a href="/authors">For Authors</a> <a href="/reviewers">For Reviewers</a> <a href="/editors">For Editors</a> <a href="/librarians">For Librarians</a> <a href="/publishing_services">For Publishers</a> <a href="/societies">For Societies</a> <a href="/conference_organizers">For Conference Organizers</a> </div> <div style="width: 100%; max-width: 250px; float: left;"> <a href="/openaccess">Open Access Policy</a> <a href="/ioap">Institutional Open Access Program</a> <a href="/special_issues_guidelines">Special Issues Guidelines</a> <a href="/editorial_process">Editorial Process</a> <a href="/ethics">Research and Publication Ethics</a> <a href="/apc">Article Processing Charges</a> <a href="/awards">Awards</a> <a href="/testimonials">Testimonials</a> </div> </div> </div> </div> <a href="/authors/english"> <h2>Editing Services</h2> </a> <div class="content__container " > <div class="custom-accordion-for-small-screen-link " > <h2>Initiatives</h2> </div> <div class="target-item custom-accordion-for-small-screen-content show-for-medium-up"> <div class="menu-container__links"> <div style="width: 100%; float: left;"> <a href="https://sciforum.net" target="_blank" rel="noopener noreferrer">Sciforum</a> <a href="https://www.mdpi.com/books" target="_blank" rel="noopener noreferrer">MDPI Books</a> <a href="https://www.preprints.org" target="_blank" rel="noopener noreferrer">Preprints.org</a> <a href="https://www.scilit.net" target="_blank" rel="noopener noreferrer">Scilit</a> <a href="https://sciprofiles.com" target="_blank" rel="noopener noreferrer">SciProfiles</a> <a href="https://encyclopedia.pub" target="_blank" rel="noopener noreferrer">Encyclopedia</a> <a href="https://jams.pub" target="_blank" rel="noopener noreferrer">JAMS</a> <a href="/about/proceedings">Proceedings Series</a> </div> </div> </div> </div> <div class="content__container " > <div class="custom-accordion-for-small-screen-link " > <h2>About</h2> </div> <div class="target-item custom-accordion-for-small-screen-content show-for-medium-up"> <div class="menu-container__links"> <div style="width: 100%; float: left;"> <a href="/about">Overview</a> <a href="/about/contact">Contact</a> <a href="https://careers.mdpi.com" target="_blank" rel="noopener noreferrer">Careers</a> <a href="/about/announcements">News</a> <a href="/about/press">Press</a> <a href="http://blog.mdpi.com/" target="_blank" rel="noopener noreferrer">Blog</a> </div> </div> </div> </div> </div> <div class="menu-container__buttons"> <a class="button UA_SignInUpButton" href="/user/login">Sign In / Sign Up</a> </div> </div> </div> <div id="captchaModal" class="reveal-modal reveal-modal-new reveal-modal-new--small" data-reveal aria-label="Captcha" aria-hidden="true" role="dialog"></div> <div id="actionDisabledModal" class="reveal-modal" data-reveal aria-labelledby="actionDisableModalTitle" aria-hidden="true" role="dialog" style="width: 300px;"> <h2 id="actionDisableModalTitle">Notice</h2> <form action="/email/captcha" method="post" id="emailCaptchaForm"> <div class="row"> <div id="js-action-disabled-modal-text" class="small-12 columns"> </div> <div id="js-action-disabled-modal-submit" class="small-12 columns" style="margin-top: 10px; display: none;"> You can make submissions to other journals <a href="https://susy.mdpi.com/user/manuscripts/upload">here</a>. </div> </div> </form> <a class="close-reveal-modal" aria-label="Close"> <i class="material-icons">clear</i> </a> </div> <div id="rssNotificationModal" class="reveal-modal reveal-modal-new" data-reveal aria-labelledby="rssNotificationModalTitle" aria-hidden="true" role="dialog"> <div class="row"> <div class="small-12 columns"> <h2 id="rssNotificationModalTitle">Notice</h2> <p> You are accessing a machine-readable page. In order to be human-readable, please install an RSS reader. </p> </div> </div> <div class="row"> <div class="small-12 columns"> <a class="button button--color js-rss-notification-confirm">Continue</a> <a class="button button--grey" onclick="$(this).closest('.reveal-modal').find('.close-reveal-modal').click(); return false;">Cancel</a> </div> </div> <a class="close-reveal-modal" aria-label="Close"> <i class="material-icons">clear</i> </a> </div> <div id="drop-article-label-openaccess" class="f-dropdown medium" data-dropdown-content aria-hidden="true" tabindex="-1"> <p> All articles published by MDPI are made immediately available worldwide under an open access license. No special permission is required to reuse all or part of the article published by MDPI, including figures and tables. For articles published under an open access Creative Common CC BY license, any part of the article may be reused without permission provided that the original article is clearly cited. For more information, please refer to <a href="https://www.mdpi.com/openaccess">https://www.mdpi.com/openaccess</a>. </p> </div> <div id="drop-article-label-feature" class="f-dropdown medium" data-dropdown-content aria-hidden="true" tabindex="-1"> <p> Feature papers represent the most advanced research with significant potential for high impact in the field. A Feature Paper should be a substantial original Article that involves several techniques or approaches, provides an outlook for future research directions and describes possible research applications. </p> <p> Feature papers are submitted upon individual invitation or recommendation by the scientific editors and must receive positive feedback from the reviewers. </p> </div> <div id="drop-article-label-choice" class="f-dropdown medium" data-dropdown-content aria-hidden="true" tabindex="-1"> <p> Editor’s Choice articles are based on recommendations by the scientific editors of MDPI journals from around the world. Editors select a small number of articles recently published in the journal that they believe will be particularly interesting to readers, or important in the respective research area. The aim is to provide a snapshot of some of the most exciting work published in the various research areas of the journal. <div style="margin-top: -10px;"> <div id="drop-article-label-choice-journal-link" style="display: none; margin-top: -10px; padding-top: 10px;"> </div> </div> </p> </div> <div id="drop-article-label-resubmission" class="f-dropdown medium" data-dropdown-content aria-hidden="true" tabindex="-1"> <p> Original Submission Date Received: <span id="drop-article-label-resubmission-date"></span>. </p> </div> <div id="container"> <noscript> <div id="no-javascript"> You seem to have javascript disabled. Please note that many of the page functionalities won't work as expected without javascript enabled. </div> </noscript> <div class="fixed"> <nav class="tab-bar show-for-medium-down"> <div class="row full-width collapse"> <div class="medium-3 small-4 columns"> <a href="/"> <img class="full-size-menu__mdpi-logo" src="https://pub.mdpi-res.com/img/design/mdpi-pub-logo-black-small1.svg?da3a8dcae975a41c?1732615622" style="width: 64px;" title="MDPI Open Access Journals"> </a> </div> <div class="medium-3 small-4 columns right-aligned"> <div class="show-for-medium-down"> <a href="#" style="display: none;"> <i class="material-icons" onclick="$('#menuModal').foundation('reveal', 'close'); return false;">clear</i> </a> <a class="js-toggle-desktop-layout-link" title="Toggle desktop layout" style="display: none;" href="/toggle_desktop_layout_cookie"> <i class="material-icons">zoom_out_map</i> </a> <a href="#" class="js-open-small-search open-small-search"> <i class="material-icons show-for-small only">search</i> </a> <a title="MDPI main page" class="js-open-menu" data-reveal-id="menuModal" href="#"> <i class="material-icons">menu</i> </a> </div> </div> </div> </nav> </div> <section class="main-section"> <header> <div class="full-size-menu show-for-large-up"> <div class="row full-width"> <div class="large-1 columns"> <a href="/"> <img class="full-size-menu__mdpi-logo" src="https://pub.mdpi-res.com/img/design/mdpi-pub-logo-black-small1.svg?da3a8dcae975a41c?1732615622" title="MDPI Open Access Journals"> </a> </div> <div class="large-8 columns text-right UI_NavMenu"> <ul> <li class="menu-item"> <a href="/about/journals" data-dropdown="journals-dropdown" aria-controls="journals-dropdown" aria-expanded="false" data-options="is_hover: true; hover_timeout: 200">Journals</a> <ul id="journals-dropdown" class="f-dropdown dropdown-wrapper dropdown-wrapper__small" data-dropdown-content aria-hidden="true" tabindex="-1"> <li> <div class="row"> <div class="small-12 columns"> <ul> <li> <a href="/about/journals"> Active Journals </a> </li> <li> <a href="/about/journalfinder"> Find a Journal </a> </li> <li> <a href="/about/journals/proposal"> Journal Proposal </a> </li> <li> <a href="/about/proceedings"> Proceedings Series </a> </li> </ul> </div> </div> </li> </ul> </li> <li class="menu-item"> <a href="/topics">Topics</a> </li> <li class="menu-item"> <a href="/authors" data-dropdown="information-dropdown" aria-controls="information-dropdown" aria-expanded="false" data-options="is_hover:true; hover_timeout:200">Information</a> <ul id="information-dropdown" class="f-dropdown dropdown-wrapper" data-dropdown-content aria-hidden="true" tabindex="-1"> <li> <div class="row"> <div class="small-5 columns right-border"> <ul> <li> <a href="/authors">For Authors</a> </li> <li> <a href="/reviewers">For Reviewers</a> </li> <li> <a href="/editors">For Editors</a> </li> <li> <a href="/librarians">For Librarians</a> </li> <li> <a href="/publishing_services">For Publishers</a> </li> <li> <a href="/societies">For Societies</a> </li> <li> <a href="/conference_organizers">For Conference Organizers</a> </li> </ul> </div> <div class="small-7 columns"> <ul> <li> <a href="/openaccess">Open Access Policy</a> </li> <li> <a href="/ioap">Institutional Open Access Program</a> </li> <li> <a href="/special_issues_guidelines">Special Issues Guidelines</a> </li> <li> <a href="/editorial_process">Editorial Process</a> </li> <li> <a href="/ethics">Research and Publication Ethics</a> </li> <li> <a href="/apc">Article Processing Charges</a> </li> <li> <a href="/awards">Awards</a> </li> <li> <a href="/testimonials">Testimonials</a> </li> </ul> </div> </div> </li> </ul> </li> <li class="menu-item"> <a href="/authors/english">Editing Services</a> </li> <li class="menu-item"> <a href="/about/initiatives" data-dropdown="initiatives-dropdown" aria-controls="initiatives-dropdown" aria-expanded="false" data-options="is_hover: true; hover_timeout: 200">Initiatives</a> <ul id="initiatives-dropdown" class="f-dropdown dropdown-wrapper dropdown-wrapper__small" data-dropdown-content aria-hidden="true" tabindex="-1"> <li> <div class="row"> <div class="small-12 columns"> <ul> <li> <a href="https://sciforum.net" target="_blank" rel="noopener noreferrer"> Sciforum </a> </li> <li> <a href="https://www.mdpi.com/books" target="_blank" rel="noopener noreferrer"> MDPI Books </a> </li> <li> <a href="https://www.preprints.org" target="_blank" rel="noopener noreferrer"> Preprints.org </a> </li> <li> <a href="https://www.scilit.net" target="_blank" rel="noopener noreferrer"> Scilit </a> </li> <li> <a href="https://sciprofiles.com" target="_blank" rel="noopener noreferrer"> SciProfiles </a> </li> <li> <a href="https://encyclopedia.pub" target="_blank" rel="noopener noreferrer"> Encyclopedia </a> </li> <li> <a href="https://jams.pub" target="_blank" rel="noopener noreferrer"> JAMS </a> </li> <li> <a href="/about/proceedings"> Proceedings Series </a> </li> </ul> </div> </div> </li> </ul> </li> <li class="menu-item"> <a href="/about" data-dropdown="about-dropdown" aria-controls="about-dropdown" aria-expanded="false" data-options="is_hover: true; hover_timeout: 200">About</a> <ul id="about-dropdown" class="f-dropdown dropdown-wrapper dropdown-wrapper__small" data-dropdown-content aria-hidden="true" tabindex="-1"> <li> <div class="row"> <div class="small-12 columns"> <ul> <li> <a href="/about"> Overview </a> </li> <li> <a href="/about/contact"> Contact </a> </li> <li> <a href="https://careers.mdpi.com" target="_blank" rel="noopener noreferrer"> Careers </a> </li> <li> <a href="/about/announcements"> News </a> </li> <li> <a href="/about/press"> Press </a> </li> <li> <a href="http://blog.mdpi.com/" target="_blank" rel="noopener noreferrer"> Blog </a> </li> </ul> </div> </div> </li> </ul> </li> </ul> </div> <div class="large-3 columns text-right full-size-menu__buttons"> <div> <a class="button button--default-inversed UA_SignInUpButton" href="/user/login">Sign In / Sign Up</a> <a class="button button--default js-journal-active-only-link js-journal-active-only-submit-link UC_NavSubmitButton" href=" https://susy.mdpi.com/user/manuscripts/upload?journal=biomedicines " data-disabledmessage="new submissions are not possible.">Submit</a> </div> </div> </div> </div> <div class="header-divider"> </div> <div class="search-container hide-for-small-down row search-container__homepage-scheme"> <form id="basic_search" style="background-color: inherit !important;" class="large-12 medium-12 columns " action="/search" method="get"> <div class="row search-container__main-elements"> <div class="large-2 medium-2 small-12 columns text-right1 small-only-text-left"> <div class="show-for-medium-up"> <div class="search-input-label"> </div> </div> <span class="search-container__title">Search<span class="hide-for-medium"> for Articles</span><span class="hide-for-small">:</span></span> </div> <div class="custom-accordion-for-small-screen-content"> <div class="large-2 medium-2 small-6 columns "> <div class=""> <div class="search-input-label">Title / Keyword</div> </div> <input type="text" placeholder="Title / Keyword" id="q" tabindex="1" name="q" value="" /> </div> <div class="large-2 medium-2 small-6 columns "> <div class=""> <div class="search-input-label">Author / Affiliation / Email</div> </div> <input type="text" id="authors" placeholder="Author / Affiliation / Email" tabindex="2" name="authors" value="" /> </div> <div class="large-2 medium-2 small-6 columns "> <div class=""> <div class="search-input-label">Journal</div> </div> <select id="journal" tabindex="3" name="journal" class="chosen-select"> <option value="">All Journals</option> <option value="acoustics" > Acoustics </option> <option value="amh" > Acta Microbiologica Hellenica (AMH) </option> <option value="actuators" > Actuators </option> <option value="admsci" > Administrative Sciences </option> <option value="adolescents" > Adolescents </option> <option value="arm" > Advances in Respiratory Medicine (ARM) </option> <option value="aerobiology" > Aerobiology </option> <option value="aerospace" > Aerospace </option> <option value="agriculture" > Agriculture </option> <option value="agriengineering" > AgriEngineering </option> <option value="agrochemicals" > Agrochemicals </option> <option value="agronomy" > Agronomy </option> <option value="ai" > AI </option> <option value="air" > Air </option> <option value="algorithms" > Algorithms </option> <option value="allergies" > Allergies </option> <option value="alloys" > Alloys </option> <option value="analytica" > Analytica </option> <option value="analytics" > Analytics </option> <option value="anatomia" > Anatomia </option> <option value="anesthres" > Anesthesia Research </option> <option value="animals" > Animals </option> <option value="antibiotics" > Antibiotics </option> <option value="antibodies" > Antibodies </option> <option value="antioxidants" > Antioxidants </option> <option value="applbiosci" > Applied Biosciences </option> <option value="applmech" > Applied Mechanics </option> <option value="applmicrobiol" > Applied Microbiology </option> <option value="applnano" > Applied Nano </option> <option value="applsci" > Applied Sciences </option> <option value="asi" > Applied System Innovation (ASI) </option> <option value="appliedchem" > AppliedChem </option> <option value="appliedmath" > AppliedMath </option> <option value="aquacj" > Aquaculture Journal </option> <option value="architecture" > Architecture </option> <option value="arthropoda" > Arthropoda </option> <option value="arts" > Arts </option> <option value="astronomy" > Astronomy </option> <option value="atmosphere" > Atmosphere </option> <option value="atoms" > Atoms </option> <option value="audiolres" > Audiology Research </option> <option value="automation" > Automation </option> <option value="axioms" > Axioms </option> <option value="bacteria" > Bacteria </option> <option value="batteries" > Batteries </option> <option value="behavsci" > Behavioral Sciences </option> <option value="beverages" > Beverages </option> <option value="BDCC" > Big Data and Cognitive Computing (BDCC) </option> <option value="biochem" > BioChem </option> <option value="bioengineering" > Bioengineering </option> <option value="biologics" > Biologics </option> <option value="biology" > Biology </option> <option value="blsf" > Biology and Life Sciences Forum </option> <option value="biomass" > Biomass </option> <option value="biomechanics" > Biomechanics </option> <option value="biomed" > BioMed </option> <option value="biomedicines" selected='selected'> Biomedicines </option> <option value="biomedinformatics" > BioMedInformatics </option> <option value="biomimetics" > Biomimetics </option> <option value="biomolecules" > Biomolecules </option> <option value="biophysica" > Biophysica </option> <option value="biosensors" > Biosensors </option> <option value="biotech" > BioTech </option> <option value="birds" > Birds </option> <option value="blockchains" > Blockchains </option> <option value="brainsci" > Brain Sciences </option> <option value="buildings" > Buildings </option> <option value="businesses" > Businesses </option> <option value="carbon" > C </option> <option value="cancers" > Cancers </option> <option value="cardiogenetics" > Cardiogenetics </option> <option value="catalysts" > Catalysts </option> <option value="cells" > Cells </option> <option value="ceramics" > Ceramics </option> <option value="challenges" > Challenges </option> <option value="ChemEngineering" > ChemEngineering </option> <option value="chemistry" > Chemistry </option> <option value="chemproc" > Chemistry Proceedings </option> <option value="chemosensors" > Chemosensors </option> <option value="children" > Children </option> <option value="chips" > Chips </option> <option value="civileng" > CivilEng </option> <option value="cleantechnol" > Clean Technologies (Clean Technol.) </option> <option value="climate" > Climate </option> <option value="ctn" > Clinical and Translational Neuroscience (CTN) </option> <option value="clinbioenerg" > Clinical Bioenergetics </option> <option value="clinpract" > Clinics and Practice </option> <option value="clockssleep" > Clocks & Sleep </option> <option value="coasts" > Coasts </option> <option value="coatings" > Coatings </option> <option value="colloids" > Colloids and Interfaces </option> <option value="colorants" > Colorants </option> <option value="commodities" > Commodities </option> <option value="complications" > Complications </option> <option value="compounds" > Compounds </option> <option value="computation" > Computation </option> <option value="csmf" > Computer Sciences & Mathematics Forum </option> <option value="computers" > Computers </option> <option value="condensedmatter" > Condensed Matter </option> <option value="conservation" > Conservation </option> <option value="constrmater" > Construction Materials </option> <option value="cmd" > Corrosion and Materials Degradation (CMD) </option> <option value="cosmetics" > Cosmetics </option> <option value="covid" > COVID </option> <option value="crops" > Crops </option> <option value="cryo" > Cryo </option> <option value="cryptography" > Cryptography </option> <option value="crystals" > Crystals </option> <option value="cimb" > Current Issues in Molecular Biology (CIMB) </option> <option value="curroncol" > Current Oncology </option> <option value="dairy" > Dairy </option> <option value="data" > Data </option> <option value="dentistry" > Dentistry Journal </option> <option value="dermato" > Dermato </option> <option value="dermatopathology" > Dermatopathology </option> <option value="designs" > Designs </option> <option value="diabetology" > Diabetology </option> <option value="diagnostics" > Diagnostics </option> <option value="dietetics" > Dietetics </option> <option value="digital" > Digital </option> <option value="disabilities" > Disabilities </option> <option value="diseases" > Diseases </option> <option value="diversity" > Diversity </option> <option value="dna" > DNA </option> <option value="drones" > Drones </option> <option value="ddc" > Drugs and Drug Candidates (DDC) </option> <option value="dynamics" > Dynamics </option> <option value="earth" > Earth </option> <option value="ecologies" > Ecologies </option> <option value="econometrics" > Econometrics </option> <option value="economies" > Economies </option> <option value="education" > Education Sciences </option> <option value="electricity" > Electricity </option> <option value="electrochem" > Electrochem </option> <option value="electronicmat" > Electronic Materials </option> <option value="electronics" > Electronics </option> <option value="ecm" > Emergency Care and Medicine </option> <option value="encyclopedia" > Encyclopedia </option> <option value="endocrines" > Endocrines </option> <option value="energies" > Energies </option> <option value="esa" > Energy Storage and Applications (ESA) </option> <option value="eng" > Eng </option> <option value="engproc" > Engineering Proceedings </option> <option value="entropy" > Entropy </option> <option value="environsciproc" > Environmental Sciences Proceedings </option> <option value="environments" > Environments </option> <option value="epidemiologia" > Epidemiologia </option> <option value="epigenomes" > Epigenomes </option> <option value="ebj" > European Burn Journal (EBJ) </option> <option value="ejihpe" > European Journal of Investigation in Health, Psychology and Education (EJIHPE) </option> <option value="fermentation" > Fermentation </option> <option value="fibers" > Fibers </option> <option value="fintech" > FinTech </option> <option value="fire" > Fire </option> <option value="fishes" > Fishes </option> <option value="fluids" > Fluids </option> <option value="foods" > Foods </option> <option value="forecasting" > Forecasting </option> <option value="forensicsci" > Forensic Sciences </option> <option value="forests" > Forests </option> <option value="fossstud" > Fossil Studies </option> <option value="foundations" > Foundations </option> <option value="fractalfract" > Fractal and Fractional (Fractal Fract) </option> <option value="fuels" > Fuels </option> <option value="future" > Future </option> <option value="futureinternet" > Future Internet </option> <option value="futurepharmacol" > Future Pharmacology </option> <option value="futuretransp" > Future Transportation </option> <option value="galaxies" > Galaxies </option> <option value="games" > Games </option> <option value="gases" > Gases </option> <option value="gastroent" > Gastroenterology Insights </option> <option value="gastrointestdisord" > Gastrointestinal Disorders </option> <option value="gastronomy" > Gastronomy </option> <option value="gels" > Gels </option> <option value="genealogy" > Genealogy </option> <option value="genes" > Genes </option> <option value="geographies" > Geographies </option> <option value="geohazards" > GeoHazards </option> <option value="geomatics" > Geomatics </option> <option value="geometry" > Geometry </option> <option value="geosciences" > Geosciences </option> <option value="geotechnics" > Geotechnics </option> <option value="geriatrics" > Geriatrics </option> <option value="glacies" > Glacies </option> <option value="gucdd" > Gout, Urate, and Crystal Deposition Disease (GUCDD) </option> <option value="grasses" > Grasses </option> <option value="hardware" > Hardware </option> <option value="healthcare" > Healthcare </option> <option value="hearts" > Hearts </option> <option value="hemato" > Hemato </option> <option value="hematolrep" > Hematology Reports </option> <option value="heritage" > Heritage </option> <option value="histories" > Histories </option> <option value="horticulturae" > Horticulturae </option> <option value="hospitals" > Hospitals </option> <option value="humanities" > Humanities </option> <option value="humans" > Humans </option> <option value="hydrobiology" > Hydrobiology </option> <option value="hydrogen" > Hydrogen </option> <option value="hydrology" > Hydrology </option> <option value="hygiene" > Hygiene </option> <option value="immuno" > Immuno </option> <option value="idr" > Infectious Disease Reports </option> <option value="informatics" > Informatics </option> <option value="information" > Information </option> <option value="infrastructures" > Infrastructures </option> <option value="inorganics" > Inorganics </option> <option value="insects" > Insects </option> <option value="instruments" > Instruments </option> <option value="iic" > Intelligent Infrastructure and Construction </option> <option value="ijerph" > International Journal of Environmental Research and Public Health (IJERPH) </option> <option value="ijfs" > International Journal of Financial Studies (IJFS) </option> <option value="ijms" > International Journal of Molecular Sciences (IJMS) </option> <option value="IJNS" > International Journal of Neonatal Screening (IJNS) </option> <option value="ijpb" > International Journal of Plant Biology (IJPB) </option> <option value="ijt" > International Journal of Topology </option> <option value="ijtm" > International Journal of Translational Medicine (IJTM) </option> <option value="ijtpp" > International Journal of Turbomachinery, Propulsion and Power (IJTPP) </option> <option value="ime" > International Medical Education (IME) </option> <option value="inventions" > Inventions </option> <option value="IoT" > IoT </option> <option value="ijgi" > ISPRS International Journal of Geo-Information (IJGI) </option> <option value="J" > J </option> <option value="jal" > Journal of Ageing and Longevity (JAL) </option> <option value="jcdd" > Journal of Cardiovascular Development and Disease (JCDD) </option> <option value="jcto" > Journal of Clinical & Translational Ophthalmology (JCTO) </option> <option value="jcm" > Journal of Clinical Medicine (JCM) </option> <option value="jcs" > Journal of Composites Science (J. Compos. Sci.) </option> <option value="jcp" > Journal of Cybersecurity and Privacy (JCP) </option> <option value="jdad" > Journal of Dementia and Alzheimer's Disease (JDAD) </option> <option value="jdb" > Journal of Developmental Biology (JDB) </option> <option value="jeta" > Journal of Experimental and Theoretical Analyses (JETA) </option> <option value="jfb" > Journal of Functional Biomaterials (JFB) </option> <option value="jfmk" > Journal of Functional Morphology and Kinesiology (JFMK) </option> <option value="jof" > Journal of Fungi (JoF) </option> <option value="jimaging" > Journal of Imaging (J. Imaging) </option> <option value="jintelligence" > Journal of Intelligence (J. Intell.) </option> <option value="jlpea" > Journal of Low Power Electronics and Applications (JLPEA) </option> <option value="jmmp" > Journal of Manufacturing and Materials Processing (JMMP) </option> <option value="jmse" > Journal of Marine Science and Engineering (JMSE) </option> <option value="jmahp" > Journal of Market Access & Health Policy (JMAHP) </option> <option value="jmp" > Journal of Molecular Pathology (JMP) </option> <option value="jnt" > Journal of Nanotheranostics (JNT) </option> <option value="jne" > Journal of Nuclear Engineering (JNE) </option> <option value="ohbm" > Journal of Otorhinolaryngology, Hearing and Balance Medicine (JOHBM) </option> <option value="jop" > Journal of Parks </option> <option value="jpm" > Journal of Personalized Medicine (JPM) </option> <option value="jpbi" > Journal of Pharmaceutical and BioTech Industry (JPBI) </option> <option value="jor" > Journal of Respiration (JoR) </option> <option value="jrfm" > Journal of Risk and Financial Management (JRFM) </option> <option value="jsan" > Journal of Sensor and Actuator Networks (JSAN) </option> <option value="joma" > Journal of the Oman Medical Association (JOMA) </option> <option value="jtaer" > Journal of Theoretical and Applied Electronic Commerce Research (JTAER) </option> <option value="jvd" > Journal of Vascular Diseases (JVD) </option> <option value="jox" > Journal of Xenobiotics (JoX) </option> <option value="jzbg" > Journal of Zoological and Botanical Gardens (JZBG) </option> <option value="journalmedia" > Journalism and Media </option> <option value="kidneydial" > Kidney and Dialysis </option> <option value="kinasesphosphatases" > Kinases and Phosphatases </option> <option value="knowledge" > Knowledge </option> <option value="labmed" > LabMed </option> <option value="laboratories" > Laboratories </option> <option value="land" > Land </option> <option value="languages" > Languages </option> <option value="laws" > Laws </option> <option value="life" > Life </option> <option value="limnolrev" > Limnological Review </option> <option value="lipidology" > Lipidology </option> <option value="liquids" > Liquids </option> <option value="literature" > Literature </option> <option value="livers" > Livers </option> <option value="logics" > Logics </option> <option value="logistics" > Logistics </option> <option value="lubricants" > Lubricants </option> <option value="lymphatics" > Lymphatics </option> <option value="make" > Machine Learning and Knowledge Extraction (MAKE) </option> <option value="machines" > Machines </option> <option value="macromol" > Macromol </option> <option value="magnetism" > Magnetism </option> <option value="magnetochemistry" > Magnetochemistry </option> <option value="marinedrugs" > Marine Drugs </option> <option value="materials" > Materials </option> <option value="materproc" > Materials Proceedings </option> <option value="mca" > Mathematical and Computational Applications (MCA) </option> <option value="mathematics" > Mathematics </option> <option value="medsci" > Medical Sciences </option> <option value="msf" > Medical Sciences Forum </option> <option value="medicina" > Medicina </option> <option value="medicines" > Medicines </option> <option value="membranes" > Membranes </option> <option value="merits" > Merits </option> <option value="metabolites" > Metabolites </option> <option value="metals" > Metals </option> <option value="meteorology" > Meteorology </option> <option value="methane" > Methane </option> <option value="mps" > Methods and Protocols (MPs) </option> <option value="metrics" > Metrics </option> <option value="metrology" > Metrology </option> <option value="micro" > Micro </option> <option value="microbiolres" > Microbiology Research </option> <option value="micromachines" > Micromachines </option> <option value="microorganisms" > Microorganisms </option> <option value="microplastics" > Microplastics </option> <option value="minerals" > Minerals </option> <option value="mining" > Mining </option> <option value="modelling" > Modelling </option> <option value="mmphys" > Modern Mathematical Physics </option> <option value="molbank" > Molbank </option> <option value="molecules" > Molecules </option> <option value="mti" > Multimodal Technologies and Interaction (MTI) </option> <option value="muscles" > Muscles </option> <option value="nanoenergyadv" > Nanoenergy Advances </option> <option value="nanomanufacturing" > Nanomanufacturing </option> <option value="nanomaterials" > Nanomaterials </option> <option value="ndt" > NDT </option> <option value="network" > Network </option> <option value="neuroglia" > Neuroglia </option> <option value="neurolint" > Neurology International </option> <option value="neurosci" > NeuroSci </option> <option value="nitrogen" > Nitrogen </option> <option value="ncrna" > Non-Coding RNA (ncRNA) </option> <option value="nursrep" > Nursing Reports </option> <option value="nutraceuticals" > Nutraceuticals </option> <option value="nutrients" > Nutrients </option> <option value="obesities" > Obesities </option> <option value="oceans" > Oceans </option> <option value="onco" > Onco </option> <option value="optics" > Optics </option> <option value="oral" > Oral </option> <option value="organics" > Organics </option> <option value="organoids" > Organoids </option> <option value="osteology" > Osteology </option> <option value="oxygen" > Oxygen </option> <option value="parasitologia" > Parasitologia </option> <option value="particles" > Particles </option> <option value="pathogens" > Pathogens </option> <option value="pathophysiology" > Pathophysiology </option> <option value="pediatrrep" > Pediatric Reports </option> <option value="pets" > Pets </option> <option value="pharmaceuticals" > Pharmaceuticals </option> <option value="pharmaceutics" > Pharmaceutics </option> <option value="pharmacoepidemiology" > Pharmacoepidemiology </option> <option value="pharmacy" > Pharmacy </option> <option value="philosophies" > Philosophies </option> <option value="photochem" > Photochem </option> <option value="photonics" > Photonics </option> <option value="phycology" > Phycology </option> <option value="physchem" > Physchem </option> <option value="psf" > Physical Sciences Forum </option> <option value="physics" > Physics </option> <option value="physiologia" > Physiologia </option> <option value="plants" > Plants </option> <option value="plasma" > Plasma </option> <option value="platforms" > Platforms </option> <option value="pollutants" > Pollutants </option> <option value="polymers" > Polymers </option> <option value="polysaccharides" > Polysaccharides </option> <option value="populations" > Populations </option> <option value="poultry" > Poultry </option> <option value="powders" > Powders </option> <option value="proceedings" > Proceedings </option> <option value="processes" > Processes </option> <option value="prosthesis" > Prosthesis </option> <option value="proteomes" > Proteomes </option> <option value="psychiatryint" > Psychiatry International </option> <option value="psychoactives" > Psychoactives </option> <option value="psycholint" > Psychology International </option> <option value="publications" > Publications </option> <option value="qubs" > Quantum Beam Science (QuBS) </option> <option value="quantumrep" > Quantum Reports </option> <option value="quaternary" > Quaternary </option> <option value="radiation" > Radiation </option> <option value="reactions" > Reactions </option> <option value="realestate" > Real Estate </option> <option value="receptors" > Receptors </option> <option value="recycling" > Recycling </option> <option value="rsee" > Regional Science and Environmental Economics (RSEE) </option> <option value="religions" > Religions </option> <option value="remotesensing" > Remote Sensing </option> <option value="reports" > Reports </option> <option value="reprodmed" > Reproductive Medicine (Reprod. Med.) </option> <option value="resources" > Resources </option> <option value="rheumato" > Rheumato </option> <option value="risks" > Risks </option> <option value="robotics" > Robotics </option> <option value="ruminants" > Ruminants </option> <option value="safety" > Safety </option> <option value="sci" > Sci </option> <option value="scipharm" > Scientia Pharmaceutica (Sci. Pharm.) </option> <option value="sclerosis" > Sclerosis </option> <option value="seeds" > Seeds </option> <option value="sensors" > Sensors </option> <option value="separations" > Separations </option> <option value="sexes" > Sexes </option> <option value="signals" > Signals </option> <option value="sinusitis" > Sinusitis </option> <option value="smartcities" > Smart Cities </option> <option value="socsci" > Social Sciences </option> <option value="siuj" > Société Internationale d’Urologie Journal (SIUJ) </option> <option value="societies" > Societies </option> <option value="software" > Software </option> <option value="soilsystems" > Soil Systems </option> <option value="solar" > Solar </option> <option value="solids" > Solids </option> <option value="spectroscj" > Spectroscopy Journal </option> <option value="sports" > Sports </option> <option value="standards" > Standards </option> <option value="stats" > Stats </option> <option value="stresses" > Stresses </option> <option value="surfaces" > Surfaces </option> <option value="surgeries" > Surgeries </option> <option value="std" > Surgical Techniques Development </option> <option value="sustainability" > Sustainability </option> <option value="suschem" > Sustainable Chemistry </option> <option value="symmetry" > Symmetry </option> <option value="synbio" > SynBio </option> <option value="systems" > Systems </option> <option value="targets" > Targets </option> <option value="taxonomy" > Taxonomy </option> <option value="technologies" > Technologies </option> <option value="telecom" > Telecom </option> <option value="textiles" > Textiles </option> <option value="thalassrep" > Thalassemia Reports </option> <option value="therapeutics" > Therapeutics </option> <option value="thermo" > Thermo </option> <option value="timespace" > Time and Space </option> <option value="tomography" > Tomography </option> <option value="tourismhosp" > Tourism and Hospitality </option> <option value="toxics" > Toxics </option> <option value="toxins" > Toxins </option> <option value="transplantology" > Transplantology </option> <option value="traumacare" > Trauma Care </option> <option value="higheredu" > Trends in Higher Education </option> <option value="tropicalmed" > Tropical Medicine and Infectious Disease (TropicalMed) </option> <option value="universe" > Universe </option> <option value="urbansci" > Urban Science </option> <option value="uro" > Uro </option> <option value="vaccines" > Vaccines </option> <option value="vehicles" > Vehicles </option> <option value="venereology" > Venereology </option> <option value="vetsci" > Veterinary Sciences </option> <option value="vibration" > Vibration </option> <option value="virtualworlds" > Virtual Worlds </option> <option value="viruses" > Viruses </option> <option value="vision" > Vision </option> <option value="waste" > Waste </option> <option value="water" > Water </option> <option value="wild" > Wild </option> <option value="wind" > Wind </option> <option value="women" > Women </option> <option value="world" > World </option> <option value="wevj" > World Electric Vehicle Journal (WEVJ) </option> <option value="youth" > Youth </option> <option value="zoonoticdis" > Zoonotic Diseases </option> </select> </div> <div class="large-2 medium-2 small-6 columns "> <div class=""> <div class="search-input-label">Article Type</div> </div> <select id="article_type" tabindex="4" name="article_type" class="chosen-select"> <option value="">All Article Types</option> <option value="research-article">Article</option> <option value="review-article">Review</option> <option value="rapid-communication">Communication</option> <option value="editorial">Editorial</option> <option value="abstract">Abstract</option> <option value="book-review">Book Review</option> <option value="brief-communication">Brief Communication</option> <option value="brief-report">Brief Report</option> <option value="case-report">Case Report</option> <option value="clinicopathological-challenge">Clinicopathological Challenge</option> <option value="article-commentary">Comment</option> <option value="commentary">Commentary</option> <option value="concept-paper">Concept Paper</option> <option value="conference-report">Conference Report</option> <option value="correction">Correction</option> <option value="creative">Creative</option> <option value="data-descriptor">Data Descriptor</option> <option value="discussion">Discussion</option> <option value="Entry">Entry</option> <option value="essay">Essay</option> <option value="expression-of-concern">Expression of Concern</option> <option value="extended-abstract">Extended Abstract</option> <option value="field-guide">Field Guide</option> <option value="guidelines">Guidelines</option> <option value="hypothesis">Hypothesis</option> <option value="interesting-image">Interesting Images</option> <option value="letter">Letter</option> <option value="books-received">New Book Received</option> <option value="obituary">Obituary</option> <option value="opinion">Opinion</option> <option value="perspective">Perspective</option> <option value="proceedings">Proceeding Paper</option> <option value="project-report">Project Report</option> <option value="protocol">Protocol</option> <option value="registered-report">Registered Report</option> <option value="reply">Reply</option> <option value="retraction">Retraction</option> <option value="note">Short Note</option> <option value="study-protocol">Study Protocol</option> <option value="systematic_review">Systematic Review</option> <option value="technical-note">Technical Note</option> <option value="tutorial">Tutorial</option> <option value="viewpoint">Viewpoint</option> </select> </div> <div class="large-1 medium-1 small-6 end columns small-push-6 medium-reset-order large-reset-order js-search-collapsed-button-container"> <div class="search-input-label"> </div> <input type="submit" id="search" value="Search" class="button button--dark button--full-width searchButton1 US_SearchButton" tabindex="12"> </div> <div class="large-1 medium-1 small-6 end columns large-text-left small-only-text-center small-pull-6 medium-reset-order large-reset-order js-search-collapsed-link-container"> <div class="search-input-label"> </div> <a class="main-search-clear search-container__link" href="#" onclick="openAdvanced(''); return false;">Advanced<span class="show-for-small-only"> Search</span></a> </div> </div> </div> <div class="search-container__advanced" style="margin-top: 0; padding-top: 0px; background-color: inherit; color: inherit;"> <div class="row"> <div class="large-2 medium-2 columns show-for-medium-up"> </div> <div class="large-2 medium-2 small-6 columns "> <div class=""> <div class="search-input-label">Section</div> </div> <select id="section" tabindex="5" name="section" class="chosen-select"> <option value=""></option> </select> </div> <div class="large-2 medium-2 small-6 columns "> <div class=""> <div class="search-input-label">Special Issue</div> </div> <select id="special_issue" tabindex="6" name="special_issue" class="chosen-select"> <option value=""></option> </select> </div> <div class="large-1 medium-1 small-6 end columns "> <div class="search-input-label">Volume</div> <input type="text" id="volume" tabindex="7" name="volume" placeholder="..." value="12" /> </div> <div class="large-1 medium-1 small-6 end columns "> <div class="search-input-label">Issue</div> <input type="text" id="issue" tabindex="8" name="issue" placeholder="..." value="10" /> </div> <div class="large-1 medium-1 small-6 end columns "> <div class="search-input-label">Number</div> <input type="text" id="number" tabindex="9" name="number" placeholder="..." value="" /> </div> <div class="large-1 medium-1 small-6 end columns "> <div class="search-input-label">Page</div> <input type="text" id="page" tabindex="10" name="page" placeholder="..." value="" /> </div> <div class="large-1 medium-1 small-6 columns small-push-6 medium-reset order large-reset-order medium-reset-order js-search-expanded-button-container"></div> <div class="large-1 medium-1 small-6 columns large-text-left small-only-text-center small-pull-6 medium-reset-order large-reset-order js-search-expanded-link-container"></div> </div> </div> </form> <form id="advanced-search" class="large-12 medium-12 columns"> <div class="search-container__advanced"> <div id="advanced-search-template" class="row advanced-search-row"> <div class="large-2 medium-2 small-12 columns show-for-medium-up"> </div> <div class="large-2 medium-2 small-3 columns connector-div"> <div class="search-input-label"><span class="show-for-medium-up">Logical Operator</span><span class="show-for-small">Operator</span></div> <select class="connector"> <option value="and">AND</option> <option value="or">OR</option> </select> </div> <div class="large-3 medium-3 small-6 columns search-text-div"> <div class="search-input-label">Search Text</div> <input type="text" class="search-text" placeholder="Search text"> </div> <div class="large-2 medium-2 small-6 large-offset-0 medium-offset-0 small-offset-3 columns search-field-div"> <div class="search-input-label">Search Type</div> <select class="search-field"> <option value="all">All fields</option> <option value="title">Title</option> <option value="abstract">Abstract</option> <option value="keywords">Keywords</option> <option value="authors">Authors</option> <option value="affiliations">Affiliations</option> <option value="doi">Doi</option> <option value="full_text">Full Text</option> <option value="references">References</option> </select> </div> <div class="large-1 medium-1 small-3 columns"> <div class="search-input-label"> </div> <div class="search-action-div"> <div class="search-plus"> <i class="material-icons">add_circle_outline</i> </div> </div> <div class="search-action-div"> <div class="search-minus"> <i class="material-icons">remove_circle_outline</i> </div> </div> </div> <div class="large-1 medium-1 small-6 large-offset-0 medium-offset-0 small-offset-3 end columns"> <div class="search-input-label"> </div> <input class="advanced-search-button button button--dark search-submit" type="submit" value="Search"> </div> <div class="large-1 medium-1 small-6 end columns show-for-medium-up"></div> </div> </div> </form> </div> <div class="header-divider"> </div> <div class="breadcrumb row full-row"> <div class="breadcrumb__element"> <a href="/about/journals">Journals</a> </div> <div class="breadcrumb__element"> <a href="/journal/biomedicines">Biomedicines</a> </div> <div class="breadcrumb__element"> <a href="/2227-9059/12">Volume 12</a> </div> <div class="breadcrumb__element"> <a href="/2227-9059/12/10">Issue 10</a> </div> <div class="breadcrumb__element"> <a href="#">10.3390/biomedicines12102216</a> </div> </div> </header> <div id="main-content" class=""> <div class="row full-width row-fixed-left-column"> <div id="left-column" class="content__column large-3 medium-3 small-12 columns"> <div class="content__container"> <a href="/journal/biomedicines"> <img src="https://pub.mdpi-res.com/img/journals/biomedicines-logo.png?8600e93ff98dbf14" alt="biomedicines-logo" title="Biomedicines" style="max-height: 60px; margin: 0 0 0 0;"> </a> <div class="generic-item no-border"> <a class="button button--color button--full-width js-journal-active-only-link js-journal-active-only-submit-link UC_ArticleSubmitButton" href="https://susy.mdpi.com/user/manuscripts/upload?form%5Bjournal_id%5D%3D158" data-disabledmessage="creating new submissions is not possible."> Submit to this Journal </a> <a class="button button--color button--full-width js-journal-active-only-link UC_ArticleReviewButton" href="https://susy.mdpi.com/volunteer/journals/review" data-disabledmessage="volunteering as journal reviewer is not possible."> Review for this Journal </a> <a class="button button--color-inversed button--color-journal button--full-width js-journal-active-only-link UC_ArticleEditIssueButton" href="/journalproposal/sendproposalspecialissue/biomedicines" data-path="/2227-9059/12/10/2216" data-disabledmessage="proposing new special issue is not possible."> Propose a Special Issue </a> </div> <div class="generic-item link-article-menu show-for-small"> <a href="#" class="link-article-menu show-for-small"> <span class="closed">►</span> <span class="open" style="display: none;">▼</span> Article Menu </a> </div> <div class="hide-small-down-initially UI_ArticleMenu"> <div class="generic-item"> <h2>Article Menu</h2> </div> <ul class="accordion accordion__menu" data-accordion data-options="multi_expand:true;toggleable: true"> <li class="accordion-navigation"> <a href="#academic_editors" class="accordion__title">Academic Editors</a> <div id="academic_editors" class="content active"> <div class="academic-editor-container " title="Department of Tumor Pathology, Graduate School of Medicine, Gifu University, Gifu 501-1194, Japan"> <div class="sciprofiles-link" style="display: inline-block"> <div class="sciprofiles-link__link"> <img class="sciprofiles-link__image" src="https://pub.mdpi-res.com/bundles/mdpisciprofileslink/img/unknown-user.png?1732615622" style="width: auto; height: 16px; border-radius: 50%;"> <span class="sciprofiles-link__name" style="line-height: 36px;">Akira Hara</span> </div> </div> </div> <div class="academic-editor-container " title="Department of Neurosciences, School of Medicine, University of Naples “Federico II”, Via Sergio Pansini 5, 80131 Naples, Italy"> <div class="sciprofiles-link" style="display: inline-block"><a class="sciprofiles-link__link" href="https://sciprofiles.com/profile/921561?utm_source=mdpi.com&utm_medium=website&utm_campaign=avatar_name" target="_blank" rel="noopener noreferrer"><img class="sciprofiles-link__image" src="/bundles/mdpisciprofileslink/img/unknown-user.png" style="width: auto; height: 16px; border-radius: 50%;"><span class="sciprofiles-link__name">Carmela Matrone</span></a></div> </div> </div> </li> <li class="accordion-direct-link"> <a href="/2227-9059/12/10/2216/scifeed_display" data-reveal-id="scifeed-modal" data-reveal-ajax="true">Subscribe SciFeed</a> </li> <li class="accordion-direct-link js-article-similarity-container" style="display: none"> <a href="#" class="js-similarity-related-articles">Recommended Articles</a> </li> <li class="accordion-direct-link"> <a href="/article/1488137/author-biographies" data-reveal-id="author-biographies-modal" data-reveal-ajax="true">Author Biographies</a> </li> <li class="accordion-navigation"> <a href="#related" class="accordion__title">Related Info Links</a> <div id="related" class="content UI_ArticleMenu_RelatedLinks"> <ul> <li class="li-link"> <a href="http://www.ncbi.nlm.nih.gov/sites/entrez/39457529" target="_blank" rel="noopener noreferrer">PubMed/Medline</a> </li> <li class="li-link"> <a href="https://scholar.google.com/scholar?q=Kalata%20B1%20Enhances%20Temozolomide%20Toxicity%20to%20Glioblastoma%20Cells" target="_blank" rel="noopener noreferrer">Google Scholar</a> </li> </ul> </div> </li> <li class="accordion-navigation"> <a href="#authors" class="accordion__title">More by Authors Links</a> <div id="authors" class="content UI_ArticleMenu_AuthorsLinks"> <ul class="side-menu-ul"> <li> <a class="expand" onclick='$(this).closest("li").next("div").toggle(); return false;'>on DOAJ</a> </li> <div id="AuthorDOAJExpand" style="display:none;"> <ul class="submenu"> <li class="li-link"> <a href='http://doaj.org/search/articles?source=%7B%22query%22%3A%7B%22query_string%22%3A%7B%22query%22%3A%22%5C%22Samantha%20L.%20Gerlach%5C%22%22%2C%22default_operator%22%3A%22AND%22%2C%22default_field%22%3A%22bibjson.author.name%22%7D%7D%7D' target="_blank" rel="noopener noreferrer">Gerlach, S. L.</a> <li> </li> <li class="li-link"> <a href='http://doaj.org/search/articles?source=%7B%22query%22%3A%7B%22query_string%22%3A%7B%22query%22%3A%22%5C%22James%20S.%20Metcalf%5C%22%22%2C%22default_operator%22%3A%22AND%22%2C%22default_field%22%3A%22bibjson.author.name%22%7D%7D%7D' target="_blank" rel="noopener noreferrer">Metcalf, J. S.</a> <li> </li> <li class="li-link"> <a href='http://doaj.org/search/articles?source=%7B%22query%22%3A%7B%22query_string%22%3A%7B%22query%22%3A%22%5C%22Rachael%20A.%20Dunlop%5C%22%22%2C%22default_operator%22%3A%22AND%22%2C%22default_field%22%3A%22bibjson.author.name%22%7D%7D%7D' target="_blank" rel="noopener noreferrer">Dunlop, R. A.</a> <li> </li> <li class="li-link"> <a href='http://doaj.org/search/articles?source=%7B%22query%22%3A%7B%22query_string%22%3A%7B%22query%22%3A%22%5C%22Sandra%20Anne%20Banack%5C%22%22%2C%22default_operator%22%3A%22AND%22%2C%22default_field%22%3A%22bibjson.author.name%22%7D%7D%7D' target="_blank" rel="noopener noreferrer">Banack, S. Anne</a> <li> </li> <li class="li-link"> <a href='http://doaj.org/search/articles?source=%7B%22query%22%3A%7B%22query_string%22%3A%7B%22query%22%3A%22%5C%22Cheenou%20Her%5C%22%22%2C%22default_operator%22%3A%22AND%22%2C%22default_field%22%3A%22bibjson.author.name%22%7D%7D%7D' target="_blank" rel="noopener noreferrer">Her, C.</a> <li> </li> <li class="li-link"> <a href='http://doaj.org/search/articles?source=%7B%22query%22%3A%7B%22query_string%22%3A%7B%22query%22%3A%22%5C%22Viswanathan%20V.%20Krishnan%5C%22%22%2C%22default_operator%22%3A%22AND%22%2C%22default_field%22%3A%22bibjson.author.name%22%7D%7D%7D' target="_blank" rel="noopener noreferrer">Krishnan, V. V.</a> <li> </li> <li class="li-link"> <a href='http://doaj.org/search/articles?source=%7B%22query%22%3A%7B%22query_string%22%3A%7B%22query%22%3A%22%5C%22Ulf%20G%C3%B6ransson%5C%22%22%2C%22default_operator%22%3A%22AND%22%2C%22default_field%22%3A%22bibjson.author.name%22%7D%7D%7D' target="_blank" rel="noopener noreferrer">Göransson, U.</a> <li> </li> <li class="li-link"> <a href='http://doaj.org/search/articles?source=%7B%22query%22%3A%7B%22query_string%22%3A%7B%22query%22%3A%22%5C%22Sunithi%20Gunasekera%5C%22%22%2C%22default_operator%22%3A%22AND%22%2C%22default_field%22%3A%22bibjson.author.name%22%7D%7D%7D' target="_blank" rel="noopener noreferrer">Gunasekera, S.</a> <li> </li> <li class="li-link"> <a href='http://doaj.org/search/articles?source=%7B%22query%22%3A%7B%22query_string%22%3A%7B%22query%22%3A%22%5C%22Blazej%20Slazak%5C%22%22%2C%22default_operator%22%3A%22AND%22%2C%22default_field%22%3A%22bibjson.author.name%22%7D%7D%7D' target="_blank" rel="noopener noreferrer">Slazak, B.</a> <li> </li> <li class="li-link"> <a href='http://doaj.org/search/articles?source=%7B%22query%22%3A%7B%22query_string%22%3A%7B%22query%22%3A%22%5C%22Paul%20Alan%20Cox%5C%22%22%2C%22default_operator%22%3A%22AND%22%2C%22default_field%22%3A%22bibjson.author.name%22%7D%7D%7D' target="_blank" rel="noopener noreferrer">Cox, P. Alan</a> <li> </li> </ul> </div> <li> <a class="expand" onclick='$(this).closest("li").next("div").toggle(); return false;'>on Google Scholar</a> </li> <div id="AuthorGoogleExpand" style="display:none;"> <ul class="submenu"> <li class="li-link"> <a href="https://scholar.google.com/scholar?q=Samantha%20L.%20Gerlach" target="_blank" rel="noopener noreferrer">Gerlach, S. L.</a> <li> </li> <li class="li-link"> <a href="https://scholar.google.com/scholar?q=James%20S.%20Metcalf" target="_blank" rel="noopener noreferrer">Metcalf, J. S.</a> <li> </li> <li class="li-link"> <a href="https://scholar.google.com/scholar?q=Rachael%20A.%20Dunlop" target="_blank" rel="noopener noreferrer">Dunlop, R. A.</a> <li> </li> <li class="li-link"> <a href="https://scholar.google.com/scholar?q=Sandra%20Anne%20Banack" target="_blank" rel="noopener noreferrer">Banack, S. Anne</a> <li> </li> <li class="li-link"> <a href="https://scholar.google.com/scholar?q=Cheenou%20Her" target="_blank" rel="noopener noreferrer">Her, C.</a> <li> </li> <li class="li-link"> <a href="https://scholar.google.com/scholar?q=Viswanathan%20V.%20Krishnan" target="_blank" rel="noopener noreferrer">Krishnan, V. V.</a> <li> </li> <li class="li-link"> <a href="https://scholar.google.com/scholar?q=Ulf%20G%C3%B6ransson" target="_blank" rel="noopener noreferrer">Göransson, U.</a> <li> </li> <li class="li-link"> <a href="https://scholar.google.com/scholar?q=Sunithi%20Gunasekera" target="_blank" rel="noopener noreferrer">Gunasekera, S.</a> <li> </li> <li class="li-link"> <a href="https://scholar.google.com/scholar?q=Blazej%20Slazak" target="_blank" rel="noopener noreferrer">Slazak, B.</a> <li> </li> <li class="li-link"> <a href="https://scholar.google.com/scholar?q=Paul%20Alan%20Cox" target="_blank" rel="noopener noreferrer">Cox, P. Alan</a> <li> </li> </ul> </div> <li> <a class="expand" onclick='$(this).closest("li").next("div").toggle(); return false;'>on PubMed</a> </li> <div id="AuthorPubMedExpand" style="display:none;"> <ul class="submenu"> <li class="li-link"> <a href="http://www.pubmed.gov/?cmd=Search&term=Samantha%20L.%20Gerlach" target="_blank" rel="noopener noreferrer">Gerlach, S. L.</a> <li> </li> <li class="li-link"> <a href="http://www.pubmed.gov/?cmd=Search&term=James%20S.%20Metcalf" target="_blank" rel="noopener noreferrer">Metcalf, J. S.</a> <li> </li> <li class="li-link"> <a href="http://www.pubmed.gov/?cmd=Search&term=Rachael%20A.%20Dunlop" target="_blank" rel="noopener noreferrer">Dunlop, R. A.</a> <li> </li> <li class="li-link"> <a href="http://www.pubmed.gov/?cmd=Search&term=Sandra%20Anne%20Banack" target="_blank" rel="noopener noreferrer">Banack, S. Anne</a> <li> </li> <li class="li-link"> <a href="http://www.pubmed.gov/?cmd=Search&term=Cheenou%20Her" target="_blank" rel="noopener noreferrer">Her, C.</a> <li> </li> <li class="li-link"> <a href="http://www.pubmed.gov/?cmd=Search&term=Viswanathan%20V.%20Krishnan" target="_blank" rel="noopener noreferrer">Krishnan, V. V.</a> <li> </li> <li class="li-link"> <a href="http://www.pubmed.gov/?cmd=Search&term=Ulf%20G%C3%B6ransson" target="_blank" rel="noopener noreferrer">Göransson, U.</a> <li> </li> <li class="li-link"> <a href="http://www.pubmed.gov/?cmd=Search&term=Sunithi%20Gunasekera" target="_blank" rel="noopener noreferrer">Gunasekera, S.</a> <li> </li> <li class="li-link"> <a href="http://www.pubmed.gov/?cmd=Search&term=Blazej%20Slazak" target="_blank" rel="noopener noreferrer">Slazak, B.</a> <li> </li> <li class="li-link"> <a href="http://www.pubmed.gov/?cmd=Search&term=Paul%20Alan%20Cox" target="_blank" rel="noopener noreferrer">Cox, P. Alan</a> <li> </li> </ul> </div> </ul> </div> </li> </ul> <span style="display:none" id="scifeed_hidden_flag"></span> <span style="display:none" id="scifeed_subscribe_url">/ajax/scifeed/subscribe</span> </div> </div> <div class="content__container responsive-moving-container large medium active hidden" data-id="article-counters"> <div id="counts-wrapper" class="row generic-item no-border" data-equalizer> <div id="js-counts-wrapper__views" class="small-12 hide columns count-div-container"> <a href="#metrics" > <div class="count-div" data-equalizer-watch> <span class="name">Article Views</span> <span class="count view-number"></span> </div> </a> </div> <div id="js-counts-wrapper__citations" class="small-12 columns hide count-div-container"> <a href="#metrics" > <div class="count-div" data-equalizer-watch> <span class="name">Citations</span> <span class="count citations-number Var_ArticleMaxCitations">-</span> </div> </a> </div> </div> </div> <div class="content__container"> <div class="hide-small-down-initially"> <ul class="accordion accordion__menu" data-accordion data-options="multi_expand:true;toggleable: true"> <li class="accordion-navigation"> <a href="#table_of_contents" class="accordion__title">Table of Contents</a> <div id="table_of_contents" class="content active"> <div class="menu-caption" id="html-quick-links-title"></div> </div> </li> </ul> </div> </div>  <div id="pbgrd-sky"></div> <script src="https://cdn.pbgrd.com/core-mdpi.js"></script> <style>.content__container { min-width: 300px; }</style>  </div> <div id="middle-column" class="content__column large-9 medium-9 small-12 columns end middle-bordered"> <div class="middle-column__help"> <div class="middle-column__help__fixed show-for-medium-up"> <span id="js-altmetrics-donut" href="#" target="_blank" rel="noopener noreferrer" style="display: none;"> <span data-badge-type='donut' class='altmetric-embed' data-doi='10.3390/biomedicines12102216'></span> <span>Altmetric</span> </span> <a href="#" class="UA_ShareButton" data-reveal-id="main-share-modal" title="Share"> <i class="material-icons">share</i> <span>Share</span> </a> <a href="#" data-reveal-id="main-help-modal" title="Help"> <i class="material-icons">announcement</i> <span>Help</span> </a> <a href="javascript:void(0);" data-reveal-id="cite-modal" data-counterslink = "https://www.mdpi.com/2227-9059/12/10/2216/cite" > <i class="material-icons">format_quote</i> <span>Cite</span> </a> <a href="https://sciprofiles.com/discussion-groups/public/10.3390/biomedicines12102216?utm_source=mpdi.com&utm_medium=publication&utm_campaign=discuss_in_sciprofiles" target="_blank" rel="noopener noreferrer" title="Discuss in Sciprofiles"> <i class="material-icons">question_answer</i> <span>Discuss in SciProfiles</span> </a> <a href="#" class="" data-hypothesis-trigger-endorses-tab title="Endorse"> <i data-hypothesis-endorse-trigger class="material-icons" >thumb_up</i> <div data-hypothesis-endorsement-count data-hypothesis-trigger-endorses-tab class="hypothesis-count-container"> ... </div> <span>Endorse</span> </a> <a href="#" data-hypothesis-trigger class="js-hypothesis-open UI_ArticleAnnotationsButton" title="Comment"> <i class="material-icons">textsms</i> <div data-hypothesis-annotation-count class="hypothesis-count-container"> ... </div> <span>Comment</span> </a> </div> <div id="main-help-modal" class="reveal-modal reveal-modal-new" data-reveal aria-labelledby="modalTitle" aria-hidden="true" role="dialog"> <div class="row"> <div class="small-12 columns"> <h2 style="margin: 0;">Need Help?</h2> </div> <div class="small-6 columns"> <h3>Support</h3> <p> Find support for a specific problem in the support section of our website. </p> <a target="_blank" href="/about/contactform" class="button button--color button--full-width"> Get Support </a> </div> <div class="small-6 columns"> <h3>Feedback</h3> <p> Please let us know what you think of our products and services. </p> <a target="_blank" href="/feedback/send" class="button button--color button--full-width"> Give Feedback </a> </div> <div class="small-6 columns end"> <h3>Information</h3> <p> Visit our dedicated information section to learn more about MDPI. </p> <a target="_blank" href="/authors" class="button button--color button--full-width"> Get Information </a> </div> </div> <a class="close-reveal-modal" aria-label="Close"> <i class="material-icons">clear</i> </a> </div> </div> <div class="middle-column__main "> <div class="page-highlight"> <style type="text/css"> img.review-status { width: 30px; } </style> <div id="jmolModal" class="reveal-modal" data-reveal aria-labelledby="Captcha" aria-hidden="true" role="dialog"> <h2>JSmol Viewer</h2> <div class="row"> <div class="small-12 columns text-center"> <iframe style="width: 520px; height: 520px;" frameborder="0" id="jsmol-content"></iframe> <div class="content"></div> </div> </div> <a class="close-reveal-modal" aria-label="Close"> <i class="material-icons">clear</i> </a> </div> <div itemscope itemtype="http://schema.org/ScholarlyArticle" id="abstract" class="abstract_div"> <div class="js-check-update-container"></div> <div class="html-content__container content__container content__container__combined-for-large__first" style="overflow: auto; position: inherit;"> <div class='html-profile-nav'> <div class='top-bar'> <div class='nav-sidebar-btn show-for-large-up' data-status='opened' > <i class='material-icons'>first_page</i> </div> <a id="js-button-download" class="button button--color-inversed" style="display: none;" href="/2227-9059/12/10/2216/pdf?version=1728463288" data-name="Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells" data-journal="biomedicines"> <i class="material-icons custom-download"></i> Download PDF </a> <div class='nav-btn'> <i class='material-icons'>settings</i> </div> <a href="/2227-9059/12/10/2216/reprints" id="js-button-reprints" class="button button--color-inversed"> Order Article Reprints </a> </div> <div class='html-article-menu'> <div class='html-first-step row'> <div class='html-font-family large-6 medium-6 small-12 columns'> <div class='row'> <div class='html-font-label large-4 medium-4 small-12 columns'> Font Type: </div> <div class='large-8 medium-8 small-12 columns'> <span class="html-article-menu-option"><i style='font-family:Arial, Arial, Helvetica, sans-serif;' data-fontfamily='Arial, Arial, Helvetica, sans-serif'>Arial</i></span> <span class="html-article-menu-option"><i style='font-family:Georgia1, Georgia, serif;' data-fontfamily='Georgia1, Georgia, serif'>Georgia</i></span> <span class="html-article-menu-option"><i style='font-family:Verdana, Verdana, Geneva, sans-serif;' data-fontfamily='Verdana, Verdana, Geneva, sans-serif' >Verdana</i></span> </div> </div> </div> <div class='html-font-resize large-6 medium-6 small-12 columns'> <div class='row'> <div class='html-font-label large-4 medium-4 small-12 columns'>Font Size:</div> <div class='large-8 medium-8 small-12 columns'> <span class="html-article-menu-option a1" data-percent="100">Aa</span> <span class="html-article-menu-option a2" data-percent="120">Aa</span> <span class="html-article-menu-option a3" data-percent="160">Aa</span> </div> </div> </div> </div> <div class='row'> <div class='html-line-space large-6 medium-6 small-12 columns'> <div class='row'> <div class='html-font-label large-4 medium-4 small-12 columns' >Line Spacing:</div> <div class='large-8 medium-8 small-12 columns'> <span class="html-article-menu-option a1" data-line-height="1.5em"> <i class="fa"></i> </span> <span class="html-article-menu-option a2" data-line-height="1.8em"> <i class="fa"></i> </span> <span class="html-article-menu-option a3" data-line-height="2.1em"> <i class="fa"></i> </span> </div> </div> </div> <div class='html-column-width large-6 medium-6 small-12 columns'> <div class='row'> <div class='html-font-label large-4 medium-4 small-12 columns' >Column Width:</div> <div class='large-8 medium-8 small-12 columns'> <span class="html-article-menu-option a1" data-column-width="20%"> <i class="fa"></i> </span> <span class="html-article-menu-option a2" data-column-width="10%"> <i class="fa"></i> </span> <span class="html-article-menu-option a3" data-column-width="0%"> <i class="fa"></i> </span> </div> </div> </div> </div> <div class='row'> <div class='html-font-bg large-6 medium-6 small-12 columns end'> <div class='row'> <div class='html-font-label large-4 medium-4 small-12 columns'>Background:</div> <div class='large-8 medium-8 small-12 columns'> <div class="html-article-menu-option html-nav-bg html-nav-bright" data-bg="bright"> <i class="fa fa-file-text"></i> </div> <div class="html-article-menu-option html-nav-bg html-nav-dark" data-bg="dark"> <i class="fa fa-file-text-o"></i> </div> <div class="html-article-menu-option html-nav-bg html-nav-creme" data-bg="creme"> <i class="fa fa-file-text"></i> </div> </div> </div> </div> </div> </div> </div> <article ><div class='html-article-content'> <span itemprop="publisher" content="Multidisciplinary Digital Publishing Institute"></span><span itemprop="url" content="https://www.mdpi.com/2227-9059/12/10/2216"></span> <div class="article-icons"><span class="label openaccess" data-dropdown="drop-article-label-openaccess" aria-expanded="false">Open Access</span><span class="label articletype">Article</span></div> <h1 class="title hypothesis_container" itemprop="name"> Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells </h1> <div class="art-authors hypothesis_container"> by <span class="inlineblock "><div class='profile-card-drop' data-dropdown='profile-card-drop13209502' data-options='is_hover:true, hover_timeout:5000'> Samantha L. Gerlach</div><div id="profile-card-drop13209502" data-dropdown-content class="f-dropdown content profile-card-content" aria-hidden="true" tabindex="-1"><div class="profile-card__title"><div class="sciprofiles-link" style="display: inline-block"><div class="sciprofiles-link__link"><img class="sciprofiles-link__image" src="/bundles/mdpisciprofileslink/img/unknown-user.png" style="width: auto; height: 16px; border-radius: 50%;"><span class="sciprofiles-link__name">Samantha L. Gerlach</span></div></div></div><div class="profile-card__buttons" style="margin-bottom: 10px;"><a href="https://sciprofiles.com/profile/621938?utm_source=mdpi.com&utm_medium=website&utm_campaign=avatar_name" class="button button--color-inversed" target="_blank"> SciProfiles </a><a href="https://scilit.net/scholars?q=Samantha%20L.%20Gerlach" class="button button--color-inversed" target="_blank"> Scilit </a><a href="https://www.preprints.org/search?search1=Samantha%20L.%20Gerlach&field1=authors" class="button button--color-inversed" target="_blank"> Preprints.org </a><a href="https://scholar.google.com/scholar?q=Samantha%20L.%20Gerlach" class="button button--color-inversed" target="_blank" rels="noopener noreferrer"> Google Scholar </a></div></div><sup> 1,2,*</sup><span style="display: inline; margin-left: 5px;"></span><a class="toEncode emailCaptcha visibility-hidden" data-author-id="13209502" href="/cdn-cgi/l/email-protection#634c000d074e00040a4c0f4c060e020a0f4e13110c170600170a0c0d40535352575450525553525205525152535201505052545202520552055251535252545607525552545355"><sup><i class="fa fa-envelope-o"></i></sup></a>, </span><span class="inlineblock "><div class='profile-card-drop' data-dropdown='profile-card-drop13209503' data-options='is_hover:true, hover_timeout:5000'> James S. Metcalf</div><div id="profile-card-drop13209503" data-dropdown-content class="f-dropdown content profile-card-content" aria-hidden="true" tabindex="-1"><div class="profile-card__title"><div class="sciprofiles-link" style="display: inline-block"><div class="sciprofiles-link__link"><img class="sciprofiles-link__image" src="/bundles/mdpisciprofileslink/img/unknown-user.png" style="width: auto; height: 16px; border-radius: 50%;"><span class="sciprofiles-link__name">James S. Metcalf</span></div></div></div><div class="profile-card__buttons" style="margin-bottom: 10px;"><a href="https://sciprofiles.com/profile/3616935?utm_source=mdpi.com&utm_medium=website&utm_campaign=avatar_name" class="button button--color-inversed" target="_blank"> SciProfiles </a><a href="https://scilit.net/scholars?q=James%20S.%20Metcalf" class="button button--color-inversed" target="_blank"> Scilit </a><a href="https://www.preprints.org/search?search1=James%20S.%20Metcalf&field1=authors" class="button button--color-inversed" target="_blank"> Preprints.org </a><a href="https://scholar.google.com/scholar?q=James%20S.%20Metcalf" class="button button--color-inversed" target="_blank" rels="noopener noreferrer"> Google Scholar </a></div><div class="profile-card__biography"><div class="js-profile-card__biography-item js-default-open"> Dr. James S. Metcalf began working at the Institute for Ethnomedicine in January 2013. He obtained a [...] </div><div class="js-profile-card__biography-item js-default-closed" style="display: none;"> Dr. James S. Metcalf began working at the Institute for Ethnomedicine in January 2013. He obtained a Joint Honours B.Sc. in Marine Biology and Biochemistry from the University of Wales, Bangor. He subsequently completed a Ph.D. at the University of Dundee in 2000 researching the immunodetection of cyanobacterial toxins. He has continued to examine various aspects of cyanobacterial toxins through Post-doctoral Fellowships at the University of Dundee, involving multinational projects on cyanotoxins throughout Europe, until 2006. Since that time he has continued an active collaboration with Prof. Geoffrey Codd of the University of Dundee. He currently works at the Department of Biological Sciences, Bowling Green State University. His current research interests include the analytical, toxicological, and molecular biological assessment of cyanobacterial toxins, in particular the neurotoxic amino acid, β-N-methylamino-L-alanine. </div><div><a href="#" class='js-profile-card__biography-item js-default-open'>Read more</a><a href="#" class='js-profile-card__biography-item js-default-closed' style="display: none;">Read less</a></div></div></div><sup> 2,3</sup><span style="display: inline; margin-left: 5px;"></span><a class="toEncode emailCaptcha visibility-hidden" data-author-id="13209503" href="/cdn-cgi/l/email-protection#b897dbd6dc95dbdfd197d497ddd5d9d1d495c8cad7ccdddbccd1d7d69b8888888f8ed988de89dd888188da888e8ad9888088dc898189de8c8c88de88dd89de"><sup><i class="fa fa-envelope-o"></i></sup></a>, </span><span class="inlineblock "><div class='profile-card-drop' data-dropdown='profile-card-drop13209504' data-options='is_hover:true, hover_timeout:5000'> Rachael A. Dunlop</div><div id="profile-card-drop13209504" data-dropdown-content class="f-dropdown content profile-card-content" aria-hidden="true" tabindex="-1"><div class="profile-card__title"><div class="sciprofiles-link" style="display: inline-block"><div class="sciprofiles-link__link"><img class="sciprofiles-link__image" src="/bundles/mdpisciprofileslink/img/unknown-user.png" style="width: auto; height: 16px; border-radius: 50%;"><span class="sciprofiles-link__name">Rachael A. Dunlop</span></div></div></div><div class="profile-card__buttons" style="margin-bottom: 10px;"><a href="https://sciprofiles.com/profile/646854?utm_source=mdpi.com&utm_medium=website&utm_campaign=avatar_name" class="button button--color-inversed" target="_blank"> SciProfiles </a><a href="https://scilit.net/scholars?q=Rachael%20A.%20Dunlop" class="button button--color-inversed" target="_blank"> Scilit </a><a href="https://www.preprints.org/search?search1=Rachael%20A.%20Dunlop&field1=authors" class="button button--color-inversed" target="_blank"> Preprints.org </a><a href="https://scholar.google.com/scholar?q=Rachael%20A.%20Dunlop" class="button button--color-inversed" target="_blank" rels="noopener noreferrer"> Google Scholar </a></div></div><sup> 2</sup><span style="display: inline; margin-left: 5px;"></span><a class="toEncode emailCaptcha visibility-hidden" data-author-id="13209504" href="/cdn-cgi/l/email-protection#c4eba7aaa0e9a7a3adeba8eba1a9a5ada8e9b4b6abb0a1a7b0adabaae7f4f4f5f7f3f6f5f5f5a5f5f7f5f3f5a1f7f6f5f3f4f2f5a5f5a7f5a0f5a2f5f3f5f2f5a6f5f5f5a6f5a7f5f3f1a7f5a0f4f4f5f1"><sup><i class="fa fa-envelope-o"></i></sup></a><a href="https://orcid.org/0000-0002-7816-3251" target="_blank" rel="noopener noreferrer"><img src="https://pub.mdpi-res.com/img/design/orcid.png?0465bc3812adeb52?1732615622" title="ORCID" style="position: relative; width: 13px; margin-left: 3px; max-width: 13px !important; height: auto; top: -5px;"></a>, </span><span class="inlineblock "><div class='profile-card-drop' data-dropdown='profile-card-drop13209505' data-options='is_hover:true, hover_timeout:5000'> Sandra Anne Banack</div><div id="profile-card-drop13209505" data-dropdown-content class="f-dropdown content profile-card-content" aria-hidden="true" tabindex="-1"><div class="profile-card__title"><div class="sciprofiles-link" style="display: inline-block"><div class="sciprofiles-link__link"><img class="sciprofiles-link__image" src="/bundles/mdpisciprofileslink/img/unknown-user.png" style="width: auto; height: 16px; border-radius: 50%;"><span class="sciprofiles-link__name">Sandra Anne Banack</span></div></div></div><div class="profile-card__buttons" style="margin-bottom: 10px;"><a href="https://sciprofiles.com/profile/4051?utm_source=mdpi.com&utm_medium=website&utm_campaign=avatar_name" class="button button--color-inversed" target="_blank"> SciProfiles </a><a href="https://scilit.net/scholars?q=Sandra%20Anne%20Banack" class="button button--color-inversed" target="_blank"> Scilit </a><a href="https://www.preprints.org/search?search1=Sandra%20Anne%20Banack&field1=authors" class="button button--color-inversed" target="_blank"> Preprints.org </a><a href="https://scholar.google.com/scholar?q=Sandra%20Anne%20Banack" class="button button--color-inversed" target="_blank" rels="noopener noreferrer"> Google Scholar </a></div><div class="profile-card__biography"><div class="js-profile-card__biography-item js-default-open"> Dr. Sandra Anne Banack received her Ph.D. degree in Integrative Biology from the University of in a [...] </div><div class="js-profile-card__biography-item js-default-closed" style="display: none;"> Dr. Sandra Anne Banack received her Ph.D. degree in Integrative Biology from the University of California, Berkeley in 1996. She received a Master of Science and a Bachelor of Science Degree from Brigham Young University in 1987 and 1985 respectively. She has been a professor at California State University Fullerton Department of Biological Science beginning in 1997 and during this time received faculty recognition for Outstanding Scholarship (2004, 2007), Outstanding Teaching (2005), and Outstanding Contributions to Student Learning (2002). In addition, she was honored with the 2006 College of Natural Science and Mathematics Dean’s Research Award. She has held positions as the Senior Scientist for the Institute of Ethnomedicine since 2004, a Research Associate for the National Tropical Botanical Garden from 2002–2004, and a Research Fellow with Umeå University, Sweden in 1996. She also served in the Department of Botany as a Visiting Assistant Professor at Brigham Young University from 1994 to 1997. Her research extends to biomarkers, exosomes, and diagnostic tools, revolutionizing our understanding of brain health. </div><div><a href="#" class='js-profile-card__biography-item js-default-open'>Read more</a><a href="#" class='js-profile-card__biography-item js-default-closed' style="display: none;">Read less</a></div></div></div><sup> 2</sup><span style="display: inline; margin-left: 5px;"></span><a class="toEncode emailCaptcha visibility-hidden" data-author-id="13209505" href="/cdn-cgi/l/email-protection#6d420e0309400e0a0442014208000c0401401d1f0219080e190402034e5d5d5c5f5a5e5c095c5a5d5c5c5f5e5e5c5b5d5a5c0f5c095c0e5c085c5b5c5a5c0c5c5d5c0c5c095c5b58095c0e5d5c5c59"><sup><i class="fa fa-envelope-o"></i></sup></a><a href="https://orcid.org/0000-0001-5183-2702" target="_blank" rel="noopener noreferrer"><img src="https://pub.mdpi-res.com/img/design/orcid.png?0465bc3812adeb52?1732615622" title="ORCID" style="position: relative; width: 13px; margin-left: 3px; max-width: 13px !important; height: auto; top: -5px;"></a>, </span><span class="inlineblock "><div class='profile-card-drop' data-dropdown='profile-card-drop13209506' data-options='is_hover:true, hover_timeout:5000'> Cheenou Her</div><div id="profile-card-drop13209506" data-dropdown-content class="f-dropdown content profile-card-content" aria-hidden="true" tabindex="-1"><div class="profile-card__title"><div class="sciprofiles-link" style="display: inline-block"><div class="sciprofiles-link__link"><img class="sciprofiles-link__image" src="/bundles/mdpisciprofileslink/img/unknown-user.png" style="width: auto; height: 16px; border-radius: 50%;"><span class="sciprofiles-link__name">Cheenou Her</span></div></div></div><div class="profile-card__buttons" style="margin-bottom: 10px;"><a href="https://sciprofiles.com/profile/author/Slk3VjRyWnUxNytXZGVpR0lGeU54dz09?utm_source=mdpi.com&utm_medium=website&utm_campaign=avatar_name" class="button button--color-inversed" target="_blank"> SciProfiles </a><a href="https://scilit.net/scholars?q=Cheenou%20Her" class="button button--color-inversed" target="_blank"> Scilit </a><a href="https://www.preprints.org/search?search1=Cheenou%20Her&field1=authors" class="button button--color-inversed" target="_blank"> Preprints.org </a><a href="https://scholar.google.com/scholar?q=Cheenou%20Her" class="button button--color-inversed" target="_blank" rels="noopener noreferrer"> Google Scholar </a></div></div><sup> 4,5</sup><span style="display: inline; margin-left: 5px;"></span><a class="toEncode emailCaptcha visibility-hidden" data-author-id="13209506" href="/cdn-cgi/l/email-protection#6d420e0309400e0a0442014208000c0401401d1f0219080e190402034e5d5d5d0f5b5e5d5b5c5c5f5e5c5b5d5d5c5d5d5a59095d5b5d5a5c5b"><sup><i class="fa fa-envelope-o"></i></sup></a>, </span><span class="inlineblock "><div class='profile-card-drop' data-dropdown='profile-card-drop13209507' data-options='is_hover:true, hover_timeout:5000'> Viswanathan V. Krishnan</div><div id="profile-card-drop13209507" data-dropdown-content class="f-dropdown content profile-card-content" aria-hidden="true" tabindex="-1"><div class="profile-card__title"><div class="sciprofiles-link" style="display: inline-block"><div class="sciprofiles-link__link"><img class="sciprofiles-link__image" src="/bundles/mdpisciprofileslink/img/unknown-user.png" style="width: auto; height: 16px; border-radius: 50%;"><span class="sciprofiles-link__name">Viswanathan V. Krishnan</span></div></div></div><div class="profile-card__buttons" style="margin-bottom: 10px;"><a href="https://sciprofiles.com/profile/586390?utm_source=mdpi.com&utm_medium=website&utm_campaign=avatar_name" class="button button--color-inversed" target="_blank"> SciProfiles </a><a href="https://scilit.net/scholars?q=Viswanathan%20V.%20Krishnan" class="button button--color-inversed" target="_blank"> Scilit </a><a href="https://www.preprints.org/search?search1=Viswanathan%20V.%20Krishnan&field1=authors" class="button button--color-inversed" target="_blank"> Preprints.org </a><a href="https://scholar.google.com/scholar?q=Viswanathan%20V.%20Krishnan" class="button button--color-inversed" target="_blank" rels="noopener noreferrer"> Google Scholar </a></div></div><sup> 5,6</sup><span style="display: inline; margin-left: 5px;"></span><a class="toEncode emailCaptcha visibility-hidden" data-author-id="13209507" href="/cdn-cgi/l/email-protection#456a262b216826222c6a296a2028242c296835372a312026312c2a2b667575747c73277577747d75767727757d747d74207521747c7520747d757075717170752075237420"><sup><i class="fa fa-envelope-o"></i></sup></a><a href="https://orcid.org/0000-0002-0726-950X" target="_blank" rel="noopener noreferrer"><img src="https://pub.mdpi-res.com/img/design/orcid.png?0465bc3812adeb52?1732615622" title="ORCID" style="position: relative; width: 13px; margin-left: 3px; max-width: 13px !important; height: auto; top: -5px;"></a>, </span><span class="inlineblock "><div class='profile-card-drop' data-dropdown='profile-card-drop13209508' data-options='is_hover:true, hover_timeout:5000'> Ulf Göransson</div><div id="profile-card-drop13209508" data-dropdown-content class="f-dropdown content profile-card-content" aria-hidden="true" tabindex="-1"><div class="profile-card__title"><div class="sciprofiles-link" style="display: inline-block"><div class="sciprofiles-link__link"><img class="sciprofiles-link__image" src="/bundles/mdpisciprofileslink/img/unknown-user.png" style="width: auto; height: 16px; border-radius: 50%;"><span class="sciprofiles-link__name">Ulf Göransson</span></div></div></div><div class="profile-card__buttons" style="margin-bottom: 10px;"><a href="https://sciprofiles.com/profile/2706564?utm_source=mdpi.com&utm_medium=website&utm_campaign=avatar_name" class="button button--color-inversed" target="_blank"> SciProfiles </a><a href="https://scilit.net/scholars?q=Ulf%20G%C3%B6ransson" class="button button--color-inversed" target="_blank"> Scilit </a><a href="https://www.preprints.org/search?search1=Ulf%20G%C3%B6ransson&field1=authors" class="button button--color-inversed" target="_blank"> Preprints.org </a><a href="https://scholar.google.com/scholar?q=Ulf%20G%C3%B6ransson" class="button button--color-inversed" target="_blank" rels="noopener noreferrer"> Google Scholar </a></div><div class="profile-card__biography"><div class="js-profile-card__biography-item js-default-open"> Ulf Göransson obtained his PhD in 2002 (Uppsala University, Dept Pharmacy, Div Pharmacognosy) on a [...] </div><div class="js-profile-card__biography-item js-default-closed" style="display: none;"> Ulf Göransson obtained his PhD in 2002 (Uppsala University, Dept Pharmacy, Div Pharmacognosy) on a thesis focussing on the discovery of a new family of circular plant peptides, named cyclotides. He completed his Post Doc in the Structural Biology laboratory of Prof David J. Craik at the Institute of Molecular Bioscience, University of Queensland, before returning to Uppsala in 2004. His laboratory focuses on the discovery of biologically active peptides from plants and animals, the development of novel techniques for the synthesis of ultrastable peptide structures, and the design of antimicrobial peptides. His lab maintains state-of-the-art equipment for peptide analysis, synthesis, and biological assays. His international network includes leading experts in the synthesis and structural determination of peptides. He obtained an Assistant Professorship from the Swedish Research Council (Biotechnology) in 2008, became an Associate Professor in 2011, and a Full Professor in 2014. His research has attracted substantial external funding from, e.g., the Swedish Research Council and the Swedish Foundation for Strategic Research. </div><div><a href="#" class='js-profile-card__biography-item js-default-open'>Read more</a><a href="#" class='js-profile-card__biography-item js-default-closed' style="display: none;">Read less</a></div></div></div><sup> 7</sup><span style="display: inline; margin-left: 5px;"></span><a class="toEncode emailCaptcha visibility-hidden" data-author-id="13209508" href="/cdn-cgi/l/email-protection#a08fc3cec48dc3c7c98fcc8fc5cdc1c9cc8dd0d2cfd4c5c3d4c9cfce83909091999795919395c2919291c19097919491c29096909691c191c293959193919490979198919791c391c195c29090909095c290969190"><sup><i class="fa fa-envelope-o"></i></sup></a>, </span><span class="inlineblock "><div class='profile-card-drop' data-dropdown='profile-card-drop13209509' data-options='is_hover:true, hover_timeout:5000'> Sunithi Gunasekera</div><div id="profile-card-drop13209509" data-dropdown-content class="f-dropdown content profile-card-content" aria-hidden="true" tabindex="-1"><div class="profile-card__title"><div class="sciprofiles-link" style="display: inline-block"><div class="sciprofiles-link__link"><img class="sciprofiles-link__image" src="/bundles/mdpisciprofileslink/img/unknown-user.png" style="width: auto; height: 16px; border-radius: 50%;"><span class="sciprofiles-link__name">Sunithi Gunasekera</span></div></div></div><div class="profile-card__buttons" style="margin-bottom: 10px;"><a href="https://sciprofiles.com/profile/2096099?utm_source=mdpi.com&utm_medium=website&utm_campaign=avatar_name" class="button button--color-inversed" target="_blank"> SciProfiles </a><a href="https://scilit.net/scholars?q=Sunithi%20Gunasekera" class="button button--color-inversed" target="_blank"> Scilit </a><a href="https://www.preprints.org/search?search1=Sunithi%20Gunasekera&field1=authors" class="button button--color-inversed" target="_blank"> Preprints.org </a><a href="https://scholar.google.com/scholar?q=Sunithi%20Gunasekera" class="button button--color-inversed" target="_blank" rels="noopener noreferrer"> Google Scholar </a></div></div><sup> 7</sup><span style="display: inline; margin-left: 5px;"></span><a class="toEncode emailCaptcha visibility-hidden" data-author-id="13209509" href="/cdn-cgi/l/email-protection#97b8f4f9f3baf4f0feb8fbb8f2faf6fefbbae7e5f8e3f2f4e3fef8f9b4a7a7a7a1a0a4a6f3a6f6a7a0a6f5a6f6a2f3a6a3a7a1a6f3a6a5a7a7a6a1a6afa6a1a7a6a6a5a4a4a6a2a6a5a7a6a6f2a6a6a6f6a6f4a2f3a7a1a7a1a2f3a7a7a6a1"><sup><i class="fa fa-envelope-o"></i></sup></a><a href="https://orcid.org/0000-0002-1089-4015" target="_blank" rel="noopener noreferrer"><img src="https://pub.mdpi-res.com/img/design/orcid.png?0465bc3812adeb52?1732615622" title="ORCID" style="position: relative; width: 13px; margin-left: 3px; max-width: 13px !important; height: auto; top: -5px;"></a>, </span><span class="inlineblock "><div class='profile-card-drop' data-dropdown='profile-card-drop13209510' data-options='is_hover:true, hover_timeout:5000'> Blazej Slazak</div><div id="profile-card-drop13209510" data-dropdown-content class="f-dropdown content profile-card-content" aria-hidden="true" tabindex="-1"><div class="profile-card__title"><div class="sciprofiles-link" style="display: inline-block"><div class="sciprofiles-link__link"><img class="sciprofiles-link__image" src="/bundles/mdpisciprofileslink/img/unknown-user.png" style="width: auto; height: 16px; border-radius: 50%;"><span class="sciprofiles-link__name">Blazej Slazak</span></div></div></div><div class="profile-card__buttons" style="margin-bottom: 10px;"><a href="https://sciprofiles.com/profile/author/U3UybnFrTVVScW9VRUtFRUY2cUEvVGhHUnV0ZzBUbk10SGV3aEUzMDNiaz0=?utm_source=mdpi.com&utm_medium=website&utm_campaign=avatar_name" class="button button--color-inversed" target="_blank"> SciProfiles </a><a href="https://scilit.net/scholars?q=Blazej%20Slazak" class="button button--color-inversed" target="_blank"> Scilit </a><a href="https://www.preprints.org/search?search1=Blazej%20Slazak&field1=authors" class="button button--color-inversed" target="_blank"> Preprints.org </a><a href="https://scholar.google.com/scholar?q=Blazej%20Slazak" class="button button--color-inversed" target="_blank" rels="noopener noreferrer"> Google Scholar </a></div></div><sup> 7,8</sup><span style="display: inline; margin-left: 5px;"></span><a class="toEncode emailCaptcha visibility-hidden" data-author-id="13209510" href="/cdn-cgi/l/email-protection#ba95d9d4de97d9ddd395d695dfd7dbd3d697cac8d5cedfd9ced3d5d4998a8a8adf8c888a898b828a8d8a828ed98b8b8adf8a898b828a898a8388888a8e8a898b8a8adc8a8a8ad88ade8ed98b8d8b8d8ed98b8b8a8d"><sup><i class="fa fa-envelope-o"></i></sup></a> and </span><span class="inlineblock "><div class='profile-card-drop' data-dropdown='profile-card-drop13209511' data-options='is_hover:true, hover_timeout:5000'> Paul Alan Cox</div><div id="profile-card-drop13209511" data-dropdown-content class="f-dropdown content profile-card-content" aria-hidden="true" tabindex="-1"><div class="profile-card__title"><div class="sciprofiles-link" style="display: inline-block"><div class="sciprofiles-link__link"><img class="sciprofiles-link__image" src="/bundles/mdpisciprofileslink/img/unknown-user.png" style="width: auto; height: 16px; border-radius: 50%;"><span class="sciprofiles-link__name">Paul Alan Cox</span></div></div></div><div class="profile-card__buttons" style="margin-bottom: 10px;"><a href="https://sciprofiles.com/profile/2929316?utm_source=mdpi.com&utm_medium=website&utm_campaign=avatar_name" class="button button--color-inversed" target="_blank"> SciProfiles </a><a href="https://scilit.net/scholars?q=Paul%20Alan%20Cox" class="button button--color-inversed" target="_blank"> Scilit </a><a href="https://www.preprints.org/search?search1=Paul%20Alan%20Cox&field1=authors" class="button button--color-inversed" target="_blank"> Preprints.org </a><a href="https://scholar.google.com/scholar?q=Paul%20Alan%20Cox" class="button button--color-inversed" target="_blank" rels="noopener noreferrer"> Google Scholar </a></div><div class="profile-card__biography"><div class="js-profile-card__biography-item js-default-open"> Paul Alan Cox graduated in Botany and Philosophy from Brigham Young University. As a Fulbright he in [...] </div><div class="js-profile-card__biography-item js-default-closed" style="display: none;"> Paul Alan Cox graduated in Botany and Philosophy from Brigham Young University. As a Fulbright Fellow, he read for his M.Sc. in Ecology at the University of Wales. He later received his A.M. and Ph.D. in Biology from Harvard University where he was a Danforth Fellow and a National Science Foundation Fellow. At Harvard, he was twice awarded the Bowdoin Prize in Literature. He was later awarded a D.Sc. Honoris Causa by the University of Guelph. After serving as a Miller Fellow at the University of California, Berkeley, he was named a Presidential Young Investigator by Ronald Reagan. In the same year, he was also named as a Melbourne University Research Fellow. He served as Professor and later Dean at Brigham Young University before being appointed as the King Carl XVI Gustaf Professor of Environmental Biology in Uppsala. He was awarded the E. K. Janaki Ammal Medal from India and is a foreign member of the Swedish Royal Academy of Agriculture and Forestry in Stockholm. </div><div><a href="#" class='js-profile-card__biography-item js-default-open'>Read more</a><a href="#" class='js-profile-card__biography-item js-default-closed' style="display: none;">Read less</a></div></div></div><sup> 2,*</sup><span style="display: inline; margin-left: 5px;"></span><a class="toEncode emailCaptcha visibility-hidden" data-author-id="13209511" href="/cdn-cgi/l/email-protection#16397578723b75717f397a39737b777f7a3b666479627375627f79783526262727212626232775252627232622272e27732770277227232722272f2725272f277327232373277026242721"><sup><i class="fa fa-envelope-o"></i></sup></a><a href="https://orcid.org/0000-0001-6401-2981" target="_blank" rel="noopener noreferrer"><img src="https://pub.mdpi-res.com/img/design/orcid.png?0465bc3812adeb52?1732615622" title="ORCID" style="position: relative; width: 13px; margin-left: 3px; max-width: 13px !important; height: auto; top: -5px;"></a></span> </div> <div class="nrm"></div> <span style="display:block; height:6px;"></span> <div></div> <div style="margin: 5px 0 15px 0;" class="hypothesis_container"> <div class="art-affiliations"> <div class="affiliation "> <div class="affiliation-item"><sup>1</sup></div> <div class="affiliation-name ">Department of Biology, Dillard University, New Orleans, LA 70122, USA</div> </div> <div class="affiliation "> <div class="affiliation-item"><sup>2</sup></div> <div class="affiliation-name ">Brain Chemistry Labs, Institute for Ethnomedicine, Jackson, WY 83001, USA</div> </div> <div class="affiliation "> <div class="affiliation-item"><sup>3</sup></div> <div class="affiliation-name ">Department of Biological Sciences, Bowling Green State University, Bowling Green, OH 43403, USA</div> </div> <div class="affiliation "> <div class="affiliation-item"><sup>4</sup></div> <div class="affiliation-name ">Department of Chemistry and Biochemistry, University of California, San Diego, CA 92093, USA</div> </div> <div class="affiliation "> <div class="affiliation-item"><sup>5</sup></div> <div class="affiliation-name ">Department of Chemistry and Biochemistry, California State University, Fresno, CA 93740, USA</div> </div> <div class="affiliation "> <div class="affiliation-item"><sup>6</sup></div> <div class="affiliation-name ">Department of Medical Pathology and Laboratory Medicine, University of California Davis School of Medicine, Davis, CA 95616, USA</div> </div> <div class="affiliation "> <div class="affiliation-item"><sup>7</sup></div> <div class="affiliation-name ">Pharmacognosy, Department of Pharmaceutical Biosciences, Uppsala University, Box 574, 751 23 Uppsala, Sweden</div> </div> <div class="affiliation "> <div class="affiliation-item"><sup>8</sup></div> <div class="affiliation-name ">W. Szafer Institute of Botany, Polish Academy of Sciences, 46 Lubicz, 31-512 Cracow, Poland</div> </div> <div class="affiliation"> <div class="affiliation-item"><sup>*</sup></div> <div class="affiliation-name ">Authors to whom correspondence should be addressed. </div> </div> </div> </div> <div class="bib-identity" style="margin-bottom: 10px;"> <em>Biomedicines</em> <b>2024</b>, <em>12</em>(10), 2216; <a href="https://doi.org/10.3390/biomedicines12102216">https://doi.org/10.3390/biomedicines12102216</a> </div> <div class="pubhistory" style="font-weight: bold; padding-bottom: 10px;"> <span style="display: inline-block">Submission received: 30 May 2024</span> / <span style="display: inline-block">Revised: 12 August 2024</span> / <span style="display: inline-block">Accepted: 15 August 2024</span> / <span style="display: inline-block">Published: 28 September 2024</span> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Section <a href="/journal/biomedicines/sections/neuro">Neurobiology and Clinical Neuroscience</a>)<br/> </div> <div class="highlight-box1"> <div class="download"> <a class="button button--color-inversed button--drop-down" data-dropdown="drop-download-1488137" aria-controls="drop-supplementary-1488137" aria-expanded="false"> Download <i class="material-icons">keyboard_arrow_down</i> </a> <div id="drop-download-1488137" class="f-dropdown label__btn__dropdown label__btn__dropdown--button" data-dropdown-content aria-hidden="true" tabindex="-1"> <a class="UD_ArticlePDF" href="/2227-9059/12/10/2216/pdf?version=1728463288" data-name="Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells" data-journal="biomedicines">Download PDF</a> <br/> <a id="js-pdf-with-cover-access-captcha" href="#" data-target="/2227-9059/12/10/2216/pdf-with-cover" class="accessCaptcha">Download PDF with Cover</a> <br/> <a id="js-xml-access-captcha" href="#" data-target="/2227-9059/12/10/2216/xml" class="accessCaptcha">Download XML</a> <br/> <a href="/2227-9059/12/10/2216/epub" id="epub_link">Download Epub</a> <br/> <a href="javascript:void(0);" data-reveal-id="supplementaryModal">Download Supplementary Material</a> <br/> </div> <div class="js-browse-figures" style="display: inline-block;"> <a href="#" class="button button--color-inversed margin-bottom-10 openpopupgallery UI_BrowseArticleFigures" data-target='article-popup' data-counterslink = "https://www.mdpi.com/2227-9059/12/10/2216/browse" >Browse Figures</a> </div> <div id="article-popup" class="popupgallery" style="display: inline; line-height: 200%"> <a href="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g001.png?1728463369" title=" <strong>Figure 1</strong><br/> <p>A representation of the structure and sequences of cyclotides. Panel (<b>A</b>) is a three-dimensional ribbon image of Kalata B1 (PDB 1NB1, SwissProt P56254), while panel (<b>B</b>) is a circular representation of Kalata B1. Panel (<b>C</b>) depicts a ball and stick model created in JSmol. The conserved cysteine residues are highlighted in yellow, while the disulfide bonds are depicted by the red bars in panel (<b>B</b>). Number of amino acids (AA), average molecular weights (ave. mass), and the sequence of amino acids are provided for cyclotides evaluated (panel (<b>D</b>)).</p> "> </a> <a href="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g002.png?1728463375" title=" <strong>Figure 2</strong><br/> <p>UPLC-MS of synthetic Kalata B1 (black peak) and natural Kalata B1 (red peak), each with a <span class="html-italic">m</span>/<span class="html-italic">z</span> of 964.77 and an elution time of 5.05–5.25 min.</p> "> </a> <a href="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g003.png?1728463391" title=" <strong>Figure 3</strong><br/> <p>MS/MS analysis of peptide 2 (RTCVP) with a mass of <span class="html-italic">m</span>/<span class="html-italic">z</span> 1215.9891.</p> "> </a> <a href="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g004.png?1728463393" title=" <strong>Figure 4</strong><br/> <p>The structural features of Kalata B1. (<b>A</b>) The amino acid sequence of Kalata B1. (<b>B</b>) An ensemble of averaged structure of Kalata B1 determined using NMR spectroscopy. The positions of the cysteines (labeled I through IV) and the corresponding disulfide linkages are shown. The * indicates a point of known cyclization between the glycine (C) and Asparagine (N) residues.</p> "> </a> <a href="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g005.png?1728463395" title=" <strong>Figure 5</strong><br/> <p>Effect of natural and synthetic Kalata B1 on cell viability in SH-SY5Y, U-87MG, and T-98G cells cultures. The IC<sub>50</sub> values for the cyclotides were determined using MTT assay after incubation with cells at increasing concentrations (0.1, 1, 5, and 10 μM) as compared to the control. Panel (<b>A</b>): SH-SY5Y and U-87 MG cells, and panel (<b>B</b>) T-98G and U-87 MG cells. Cultures (6000 cells per well in a 96-well plate), were treated with the cyclotide indicated, for 72 h, then MTT measured. Cell viability was calculated as a percentage of control cultures. IC<sub>50</sub> values were calculated using the Best Fit method (See <a href="#biomedicines-12-02216-t001" class="html-table">Table 1</a>). Incubations were conducted in 96-well plates in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). The results are graphed in GraphPad Prism version 9 for iOS.</p> "> </a> <a href="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g006.png?1728463397" title=" <strong>Figure 6</strong><br/> <p>Effect of increasing the concentration of naturally occurring cyclotides on cell viability in U-87 MG cells. IC<sub>50</sub> values for cyclotides were determined using the MTT assay after incubating U-87 MG cell cultures with increasing concentrations (0.1, 1.0, 5.0, and 10 μM) or with a control of DMSO only. Panel (<b>A</b>): CyO3, panel (<b>B</b>): CyO19, panel (<b>C</b>): Kalata B1, panel (<b>D</b>): Kalata B7, and panel (<b>E</b>), Vitri E. U-87 cells (6000 cells per well in a 96-well plate) were treated with cyclotides, as indicated, for 72 h, then MTT measured. Cell viability was calculated as a percentage of control cultures. IC<sub>50</sub> values were calculated using the Best Fit method (See <a href="#biomedicines-12-02216-t001" class="html-table">Table 1</a>). Incubations were conducted in 96-well plates in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). Results are graphed in GraphPad Prism version 9 for iOS.</p> "> </a> <a href="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g007.png?1728463401" title=" <strong>Figure 7</strong><br/> <p>Comparison of IC<sub>50</sub> values for natural and synthetic Kalata B1, as determined in SH-SY5Y, U-87 MG, and T-98G human cell cultures. Panel (<b>A</b>) includes SH-SY5Y and U-87 MG cells, panel (<b>B</b>) includes T-98G and U-87 MG cells, and panel (<b>C</b>) is natural Kalata B1 and synthetic Kalata B1 with concentrations of 0.1, 1, 5, and 10 μM as compared to the control. Cultures were plated in 96-well pates at 6000 cells/well, then treated with cyclotides, as indicated, for 72 h, then assayed using MTT. Cell viability was calculated as a percentage of control (where the control cells were treated with DMSO only). Incubations were conducted in 96-well plates in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). IC<sub>50</sub> values were calculated using the Best Fit method in GraphPad Prism version 8.0 (IC50 Best Fit). Results are graphed in GraphPad Prism version 8 for iOS.</p> "> </a> <a href="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g008.png?1728463403" title=" <strong>Figure 8</strong><br/> <p>Synthetic Kalata B1 increased TMZ-induced cell death in U-87 MG glioblastoma cultures. To determine if synthetic Kalata B1 (K) enhances TMZ (T)-induced cell death in U-87 MG cells, MTT was measured after incubating the cell cultures with concentrations of K (0.5, 1.0, 2.0, and 4.0 μM) and T (100, 200, 400, and 800 μM) both alone and when co-exposed to synthetic Kalata B1 and TMZ as illustrated in panels (<b>A</b>–<b>M</b>). The first dotted bars in each panel indicate the cells exposed to concentrations of synthetic Kalata B1 alone, while the second diagonal stripe bars indicate the cells exposed to TMZ alone. The third checkered bars indicate co-exposure treatments with synthetic Kalata B1 and TMZ. The cells were plated into 96-well plates (6000 cells/well) and treated with synthetic Kalata B, as indicated, for 72 h. Cell viability was measured using MTT and expressed as a percentage of control cultures (culture medium with DMSO only). Incubations were conducted in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). Results of one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons are graphed in GraphPad Prism version 9 for iOS with the following <span class="html-italic">p</span>-values: * &lt; 0.05, ** &lt; 0.01, *** &lt; 0.001, and **** &lt; 0.0001. ns = not significant.</p> "> </a> <a href="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g009.png?1728463407" title=" <strong>Figure 9</strong><br/> <p>Synthetic Kalata B1 increased TMZ-induced cell death in T-98G glioblastoma cultures. To determine if Kalata B1 (K) enhances TMZ (T)-induced cell death in T-98G cells, MTT was measured after incubating cell cultures with synthetic Kalata B1 (0.25, 0.5, 1.0, and 2.0 μM) and TMZ (T 75, 150, 300, and 600 μM) both alone and when co-exposed as illustrated in panels (<b>A</b>–<b>I</b>). The first dotted bars in each panel indicate the cells exposed to synthetic Kalata B1 alone, while the second diagonal stripe bars indicate the cells exposed to TMZ alone. The third checkered bars indicate co-exposure treatments with synthetic Kalata B1 and TMZ. The cells were plated into 96-well plates (6000 cells/well) and treated with synthetic Kalata B1, as indicated, for 72 h. Cell viability was measured using MTT and expressed as a percentage of control cultures (culture medium + DMSO only). Incubations were conducted in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). The data are expressed as mean ± standard error of the mean (SEM). The results of one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons are graphed in GraphPad Prism version 9 for iOS, with the following <span class="html-italic">p</span>-values: * &lt; 0.05, ** &lt; 0.01, *** &lt; 0.001, and **** &lt; 0.0001. ns = not significant.</p> "> </a> <a href="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g010.png?1728463412" title=" <strong>Figure 10</strong><br/> <p>Phase contrast microscopy of U-87 GM cultures treated with Kalata B1 and temozolomide. Panels are as follows: (<b>A</b>) no treatment/control; (<b>B</b>) 150 µM temozolomide (TMZ); (<b>C</b>) synthetic Kalata B1 0.5 µM; and (<b>D</b>) 150 µM TMZ + 0.5 µM synthetic Kalata B1 for 72 h. Imaged on an Olympus IX83 inverted microscope equipped with an Olympus XM10 camera and acquired in cellSens standard. Images were adjusted for exposure in Adobe Photoshop 23.4.2 using “Levels” and cropped for consistency and to enable ease of comparison. U-87 GM cells were plated at 6000 cells/well in a 96-well plate and allowed to adhere overnight prior to treatment. Images are representative of triplicate incubations. The yellow bar in the lower right-hand corner of the image is 50 µm. 20× magnification.</p> "> </a> <a href="https://pub.mdpi-res.com/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g011.png?1728463418" title=" <strong>Figure 11</strong><br/> <p>Stability of synthetic Kalata B1 and temozolomide (TMZ) in human serum over 24 h. Percentage of Kalata B1 and TMZ remaining upon incubation with human serum. The portion of serum-treated peptide remaining was determined by calculating the area of the respective elution peak in UPLC-MS and comparing it with the control without incubation. Data points are means ± SEM.</p> "> </a> </div> <a class="button button--color-inversed" href="/2227-9059/12/10/2216/notes">Versions Notes</a> </div> </div> <div class="responsive-moving-container small hidden" data-id="article-counters" style="margin-top: 15px;"></div> <div class="html-dynamic"> <section> <div class="art-abstract art-abstract-new in-tab hypothesis_container"> <p> <div><section class="html-abstract" id="html-abstract"> <h2 id="html-abstract-title">Abstract</h2><b>:</b> <div class="html-p">Glioblastoma (GBM) is the most aggressive cancer originating in the brain, but unfortunately combination treatments with resection, radiation, and chemotherapy are relatively ineffective. Therefore, novel methods of adjuvant therapy are critically needed. Cyclotides are plant-derived circular peptides that chemosensitize drug-resistant breast cancer to doxorubicin. We analyzed naturally occurring and synthetic cyclotides (Cycloviolacin O3, Cycloviolacin O19, natural Kalata B1, synthetic Kalata B1, and Vitri E) alone and in co-exposure treatments with the drug temozolomide (TMZ) in human glioblastoma cells. The cyclotides were identified by UPLC-PDA and HPLC-UV. The synthetic Kalata B1 sequence was verified with orbitrap LC-MS, and structural confirmation was provided by NMR spectroscopy. The cyclotides displayed dose-dependent cytotoxicity (IC<sub>50</sub> values 2.4–21.1 µM) both alone and as chemosensitizers of U-87 MG and T 98 cells to TMZ. In fact, a 16-fold lower concentration of TMZ (100 µM) was needed for significant cytotoxicity in U-87 MG cells co-exposed to synthetic Kalata B (0.5 µM). Similarly, a 15-fold lower concentration of TMZ (75 µM) was required for a significant reduction in cell viability in T 98 cells co-exposed to synthetic Kalata B1 (0.25 µM). Kalata B1 remained stable in human serum stability assays. The data support the assertion that cyclotides may chemosensitize glioblastoma cells to TMZ.</div> </section> <div id="html-keywords"> <div class="html-gwd-group"><div id="html-keywords-title">Keywords: </div><a href="/search?q=cyclotides">cyclotides</a>; <a href="/search?q=glioblastoma">glioblastoma</a>; <a href="/search?q=temozolomide">temozolomide</a>; <a href="/search?q=chemosensitize">chemosensitize</a>; <a href="/search?q=chemotherapy">chemotherapy</a>; <a href="/search?q=adjuvant+therapy">adjuvant therapy</a>; <a href="/search?q=Viola"><span class="html-italic">Viola</span></a></div> <div> </div> </div> </div> </p> </div> </section> </div> <div class="hypothesis_container"> <ul class="menu html-nav" data-prev-node="#html-quick-links-title"> </ul> <div class="html-body"> <section id='sec1-biomedicines-12-02216' type='intro'><h2 data-nested='1'> 1. Introduction</h2><div class='html-p'>The discovery of cyclotides dates back to 1973, when they were found to be the active peptides responsible for increasing uterine contractions and hastening labor within an ethnobotanical tea used by indigenous populations in Africa. Traditional healers of the Lulua Tribe in the Democratic Republic of the Congo boiled the aerial tissues of a plant they call Kalata-Kalata (<span class='html-italic'>Oldenlandia affinis</span> (R & S) DC, Rubiaceae) and ingested the tea to facilitate labor [<a href="#B1-biomedicines-12-02216" class="html-bibr">1</a>]. One active compound elucidated from the plant extract is Kalata B1, the prototypic cyclotide. Cyclotides can be divided into three subfamilies, including bracelet, Möbius, and trypsin inhibitors, based on their structural and functional differences [<a href="#B2-biomedicines-12-02216" class="html-bibr">2</a>]. The circular variants can be found in five plant families and are common in violets (Violaceae), while they are sporadically found in genera of the Rubiaceae family. Within plants, cyclotides contribute to host defense and have been found to have insecticidal, antibacterial, and antifungal properties [<a href="#B3-biomedicines-12-02216" class="html-bibr">3</a>,<a href="#B4-biomedicines-12-02216" class="html-bibr">4</a>,<a href="#B5-biomedicines-12-02216" class="html-bibr">5</a>,<a href="#B6-biomedicines-12-02216" class="html-bibr">6</a>]. Cyclotides are resistant to thermal, chemical, and enzymatic degradation, which makes them promising candidates for therapeutic research [<a href="#B7-biomedicines-12-02216" class="html-bibr">7</a>,<a href="#B8-biomedicines-12-02216" class="html-bibr">8</a>].</div><div class='html-p'>One mechanism of cyclotide bioactivity includes the formation of monomer to tetramer pores on the phospholipid bilayer of cells. Cyclotides target the phosphatidylethanolamine (PE)-enriched head groups of phospholipids and disrupt lipid membranes via insertion [<a href="#B5-biomedicines-12-02216" class="html-bibr">5</a>,<a href="#B7-biomedicines-12-02216" class="html-bibr">7</a>]. The use of plants containing cyclotides in ethnomedical remedies has led to purified compounds which display both anti-cancer and anti-HIV activity [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>,<a href="#B10-biomedicines-12-02216" class="html-bibr">10</a>]. We have found that cyclotides can chemosensitize HIV-infected cells, breast cancer cells, and glioblastoma cells to FDA-approved drugs [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>,<a href="#B11-biomedicines-12-02216" class="html-bibr">11</a>,<a href="#B12-biomedicines-12-02216" class="html-bibr">12</a>].</div><div class='html-p'>Glioblastoma is a devastating and lethal brain cancer in children and adults, with a median survival rate of 9–16 months post diagnosis. The challenges associated with treatment include the lack of efficacy associated with resection, radiation, and chemotherapy [<a href="#B13-biomedicines-12-02216" class="html-bibr">13</a>,<a href="#B14-biomedicines-12-02216" class="html-bibr">14</a>]. The standard post-surgical drug regimen consists of temozolomide (TMZ); however, drug resistance is often rapid and only 50% of patients respond to TMZ. One significant challenge in the clinical treatment of patients suffering from glioblastoma is that high doses for prolonged periods (up to 49 days) may be required for efficacy, which can precipitously lead to TMZ resistance. After TMZ resistance develops, there are few, if any, other treatment options available since TMZ is considered the current “gold standard” of care [<a href="#B15-biomedicines-12-02216" class="html-bibr">15</a>,<a href="#B16-biomedicines-12-02216" class="html-bibr">16</a>].</div><div class='html-p'>Combination strategies with glioblastoma treatment have been hampered, at least in part, by the infiltrative nature of the tumor cells and the resistance and recurrence due to self-renewing tumor cell types such as glioblastoma stem cells, which escape chemo-radiotherapy and proliferate following treatment [<a href="#B15-biomedicines-12-02216" class="html-bibr">15</a>]. Our previous research indicates that a combination approach using cyclotides and FDA-approved drugs may chemosensitize breast cancer cells to doxorubicin [<a href="#B11-biomedicines-12-02216" class="html-bibr">11</a>], HIV-infected cells to saquinavir and T-20 [<a href="#B12-biomedicines-12-02216" class="html-bibr">12</a>], and glioblastoma cells to TMZ [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>].</div><div class='html-p'>We evaluated five natural and one synthetic cyclotide in U-87 MG and T-98G human glioblastoma cells to evaluate their cytotoxicity alone and in combination with TMZ. Cycloviolacin O3 (CyO3) and Cycloviolacin O19 (CyO19) are bracelet cyclotides, whereas Kalata B1 (KB1), Kalata B7 (KB7), and Vitri E are Möbius cyclotides with 29–31 amino acids (<a href="#biomedicines-12-02216-f001" class="html-fig">Figure 1</a>). Additionally, we identified the sequence of synthetic Kalata B1 using orbitrap LC-MS, verified its structure with NMR spectroscopy, and evaluated its stability in human serum. The current research significantly advances existing glioblastoma studies by demonstrating, for the first time to our knowledge, the dose-dependent cytotoxicity of several natural cyclotides (CyO3, CyO19, Kalata B7, and Vitri E) against U-87 MG human glioblastoma cells and the chemosensitizing property of the synthetically derived Kalata B1.</div></section><section id='sec2-biomedicines-12-02216' type=''><h2 data-nested='1'> 2. Materials and Methods</h2><section id='sec2dot1-biomedicines-12-02216' type='methods'><h4 class='html-italic' data-nested='2'> 2.1. General Experimental Procedures</h4><div class='html-p'>American Type Culture Collection (ATCC) provided the SH-SY5Y human neuro- blastoma (ATCC CRL-2266, PD21-26), U-87 MG human glioblastoma (ATCC HTB-14), and T-98G (ATCC CRL-1690) cells, as well as Eagle’s minimal essential medium (ATCC Cat. No. 30-2003). Dulbecco’s modified Eagle’s medium/Nutrient F-12 Ham (Cat. No. D8437), the Roche cell proliferation kit I (MTT) (Cat. No. 11465007001, Roche, Basel, Switzerland), dimethyl sulfoxide (DMSO) (Cat. No. D8418), NaCl (Cat. No. S7650), ammonium bicarbonate (NH4HCO3, Cat. No. A6141), acetonitrile (AcN, Cat. No. 34851), iodoacetamide (Cat. No. A3221), endoproteinase Glu-C (Cat. No. 1140399001), human serum isolated from male AB plasma (Cat. No. H4522), urea (Cat. No. U5128), and trichloroacetic acid (Cat. No. T6399) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dithiothreitol (Cat. No. 1610610) was purchased from Bio-Rad (Hercules, CA, USA). Temozolomide was provided by Selleck Chemicals, Houston, TX, USA (Cat. No. S1237). Trypsin (Cat. No. V5280) and chymotrypsin (Cat. No. V1061) were obtained from Promega (Madison, WI, USA). C18 (EC) SPE columns (Part No. 221-0100-B) were purchased from Biotage (Uppsala, Sweden). LiChroSolv water (Cat. No. 1.15333.1000), C18 Ziptips (Ref. No. ZTC185960), and the Ultrafree MCGV centrifugal 0.22 μm filters, Durapore RVDF (Ref. No. UFC30GVNB), were obtained through Millipore-Sigma, St. Louis, MO, USA. Nylon filters (0.2 μm, Cat. No. 7402-004) were supplied by Whatman GE Healthcare Life Sciences (Cleves, OH, USA). Ethanol (EtOH, Cat. No. 22032106) and trifluoroacetic acid (Cat. No. 04902-100) were purchased from Fisher Scientific (Waltham, MA, USA), while formic acid (Cat. No. 28905) and 11 mm, 600 μL plastic crimp/snap top autosampler vials (Cat. No. C4011-11) were obtained from Thermo Fisher Scientific. The Qubit standard protein assay kit (Cat. No. Q33212) was provided by Thermo Fisher Scientific. Each chemical was cell culture or reagent grade, and all digestive enzymes were sequencing grade.</div></section><section id='sec2dot2-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 2.2. Isolation and Purification of Plant Peptides</h4><div class='html-p'>Cyclotides CyO3, CyO19, natural Kalata B1, and Vitri E were isolated using similar procedures described in a previous study [<a href="#B6-biomedicines-12-02216" class="html-bibr">6</a>]. CyO3, CyO19, and natural Kalata B1 were isolated from <span class='html-italic'>V. odorata</span> L., while Vitri E was isolated from <span class='html-italic'>V. tricolor</span> L. Plant material (leaves) of both species was bought from a commercial supplier, Alfred Galke GmbH (Gittelde, Germany). About 200 g of plant material was extracted with 2L of 60% methanol overnight. Subsequently, the extracts were diluted with water to below 20% methanol and loaded into a C18 low-pressure column (C18 spherical, 40–60 μm, 100 A, 40 g, Agela Technologies, Torrance, CA, USA) using a pump. The loaded columns were attached to a low-pressure chromatography system (Äkta FPLC, GE Healthcare, Chicago, IL, USA) operated with a flow rate of 20 mL min<sup>−1</sup> and a gradient of 10–95% ACN, 0.1% TFA, and one min fractions were collected. The fractions containing the cyclotides were identified using LCQ electrospray ion trap MS (Thermo Finnigan, San Jose, CA, USA), operated in the positive-ion mode with direct injection. The peptides were purified from their respective fractions using HPLC. Kalata B7 was isolated from <span class='html-italic'>Geophilia repens</span> (Rubiaceae) during a previous study [<a href="#B17-biomedicines-12-02216" class="html-bibr">17</a>].</div><div class='html-p'>CS Bio Co. (Menlo Park, CA, USA, 94025) synthesized 30 mg of Kalata B1 (Sample No. CS9566, Lot No. X477) with a purity of 97.51%, as analyzed by Agilent (Santa Clara, CA, USA) 1290 HPLC using a Phenomenex Luna C 18 5 µm 100 Å, 4.6 × 250 mm column (25 °C). The HPLC method involved 30–60% Buffer B for 20 min (Buffer A = 0.1% TFA in H<sub>2</sub>O and Buffer B = 0.1% TFA in ACN) with a flow rate of 1 mL/min., monitored at a wavelength of 214 nm. A linear peptide was synthesized, cyclized, and properly folded using head-to-tail lactam bonding and natural oxidation (<a href="#app1-biomedicines-12-02216" class="html-app">Figure S1</a>) and the identity was confirmed via comparison with the natural Kalata B1.</div></section><section id='sec2dot3-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 2.3. Verification of Synthetic Kalata B1</h4><section id='sec2dot3dot1-biomedicines-12-02216' type=''><h4 class='' data-nested='3'> 2.3.1. UPLC-MS Synthetic Kalata B1 Verification</h4><div class='html-p'>Purified CyO3, CyO19, natural Kalata B1, Kalata B7, and Vitri E were subjected to ultraperformance liquid chromatography coupled with photodiode array detection (UPLC-PDA) and reverse-phase high-performance liquid chromatography with UV detection (HPLC-UV). During UPCL-PDA, 5 µL injections (10% (<span class='html-italic'>v</span>/<span class='html-italic'>v</span>) ethanol (EtOH) in water) were loaded onto a C 18 LC Column (Kinetex Core-Shell) of 100 mm × 2.1 mm, 1.7 µM, 100 Å. The flow rate was maintained at 0.65 mL/min using a Waters Acquity Ultraperformance LC (Milford, MA, USA). The solvents were as follows: water + 0.1% (<span class='html-italic'>v</span>/<span class='html-italic'>v</span>) trifluoroacetic acid (A) and acetonitrile + 0.1% (<span class='html-italic'>v</span>/<span class='html-italic'>v</span>) trifluoroacetic acid. The gradient was set from 0–10% B for 1 min, then 10–60% B for 6 min, followed by 60–85% B for 2 min. The column temperature was set at 40 °C, while the sample temperature was 7 °C. During the UPLC-PDA analysis, the same gradient and temperatures were used; however, the loop was switched from partial to a full loop. The following <span class='html-italic'>m</span>/<span class='html-italic'>z</span> ratios were monitored: TMZ (195.06), synthetic and natural Kalata B1 (964.77), CyO3 (1051.80), CyO19 (1076.94), and Vitri E (975.45). Chromatograms were evaluated using Empower 3 imaging software, version 7.50.00.00.</div></section><section id='sec2dot3dot2-biomedicines-12-02216' type=''><h4 class='' data-nested='3'> 2.3.2. Orbitrap Exploris 480 Liquid Chromatography-Mass Spectrometry</h4><div class='html-p'>Synthetic Kalata B1 peptide was modified and digested with endoGluC and trypsin before being cleaned up with C18 zip tips, according to Gerlach and colleagues [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>]. The dried residue was resuspended with 20 μl Direct Q water + 0.1% (<span class='html-italic'>v</span>/<span class='html-italic'>v</span>) formic acid for mass spec analysis. The samples were analyzed using an Orbitrap Exploris 480 mass spectrometer (Thermo Scientific) and an Easy-nLC 1200 (Thermo Scientific) nano liquid chromatography system. For the LC separation, samples (1 μL) were injected onto a nanoEase C18 column (Waters MZ HSS T3 column, 100 Å, 1.8 μm, 75 μm × 150 mm) at a flow rate of 20 μL per minute in a volume of 6μL and maximum pressure of 980 bar with solvents of water (A, LiChrosolv, EMD Millipore) + 0.1% formic acid (Ricca Chemicals, Arlington, TX, USA) and 80% acetonitrile in water (LiChrosolv, EMD Millipore) + 0.1% formic acid. The samples were separated for mass spectrometric analysis using a gradient of 5–100% B over 12 min, maintaining 100% B for a further 8 min with a flow of 300 nL/min and a total run time of 20 min. The analytical column equilibration was 10 μL, with a maximum pressure of 500 bar. The flush volume was set at 100 μL. Samples were analyzed by tSIM-MS/MS, modifying several parameters to improve detection of the peptides. The source was maintained at a spray voltage of 1.4 kV and capillary temperature of 250 °C. After digestion, for the optimal detection of the two Kalata B1 peptides, the following methods were used.</div><div class='html-p'><span class='html-italic'>Peptide 1 (NGLPVCGE)</span>: A tSIM method was set up with a default charge state of 2, but it also included a charge state of 1, an isolation window of <span class='html-italic'>m</span>/<span class='html-italic'>z</span> = 2, an orbitrap resolution of 480,000, 1 microscan, an RF lens setting of 50%, and 20 dependent scans. tSIMs of 845.3821 (z = 1) and 423.1947 (z = 2) were used to isolate peptides before MS/MS. These scans were performed with an isolation window of <span class='html-italic'>m</span>/<span class='html-italic'>z</span> = 2, a stepped energy mode of 15, 30, 45%, an orbitrap resolution of 480,000, 1 microscan, and an expected LC peak width of 15. All other parameters were set at the manufacturers’ default.</div><div class='html-p'><span class='html-italic'>Peptide 2 (RTCVPWSCTCGPTNCTGGVCT)</span>: Mass spectrometric analysis was performed the same as for peptide 1 with the following changes. During the tSIM, charge states of 2 and 3 were assessed with an RF lens of 125% and tSIMs of 1215.9894 (z = 2) and 810.9954 (z = 3). The MS/MS parameters were the same as for peptide 1, with stepped energies of 20, 40, 60%. After analysis, all peptides were identified and sequenced by hand in Freestyle software version 1.7 (Thermo Scientific).</div></section><section id='sec2dot3dot3-biomedicines-12-02216' type=''><h4 class='' data-nested='3'> 2.3.3. NMR Spectroscopy</h4><div class='html-p'>Approximately 10 mg of the peptide was prepared in 600 µL in 90% H<sub>2</sub>O/10% D<sub>2</sub>O. All the NMR experiments were performed at a pH of 3.6 and a sample temperature of 303 K. The NMR experiments were performed on a JEOL 600 MHz spectrometer (Fresno State University, Fresno, CA, USA) equipped with the SuperCOOL cryogenic (liquid N<sub>2</sub>) probe with a triple-axis pulse-field gradient. The following two-dimensional (2D) NMR experiments were collected: <sup>1</sup>H-<sup>1</sup>H double quantum filtered correlation spectroscopy (DQFC) [<a href="#B18-biomedicines-12-02216" class="html-bibr">18</a>], <sup>1</sup>H-<sup>1</sup>H total correlation spectroscopy (TOCSY, 80 ms with a DIPSI spin-lock) [<a href="#B19-biomedicines-12-02216" class="html-bibr">19</a>], <sup>1</sup>H-<sup>1</sup>H nuclear Overhauser effect spectroscopy (NOESY, 150 ms), and <sup>1</sup>H-<sup>1</sup>H rotating frame nuclear Overhauser effect spectroscopy (ROESY, 200 ms with adiabatic spin-lock with a 6.0 kHz) [<a href="#B20-biomedicines-12-02216" class="html-bibr">20</a>,<a href="#B21-biomedicines-12-02216" class="html-bibr">21</a>,<a href="#B22-biomedicines-12-02216" class="html-bibr">22</a>].</div><div class='html-p'>Typically, the time domain data were collected with 4096 × 256 (t<sub>2</sub> × t<sub>1</sub>) complex points in a phase-sensitive mode, and 128 transients were collected per t<sub>1</sub> increment with a relaxation delay of 2.0 s. The chemical shifts were referenced using sodium trimethyl-silyl propane sulfonate (DSS) (TMS) as the internal standard. Three-bond <sup>1</sup>H-<sup>1</sup>H coupling constants were measured from one-dimensional spectra. NMR data were processed using a combination of JEOL Delta (zero-filled twice, sine-squared window functions, and complex Fourier transform) and saved in the Sparky [<a href="#B23-biomedicines-12-02216" class="html-bibr">23</a>,<a href="#B24-biomedicines-12-02216" class="html-bibr">24</a>] format. The integration function in Sparky was performed to obtain the volume for each NOE cross peak. Finally, the chemical shifts and NOE cross-peak volume were exported from Sparky for structure calculation.</div></section><section id='sec2dot3dot4-biomedicines-12-02216' type=''><h4 class='' data-nested='3'> 2.3.4. The Ensemble of NMR Structures</h4><div class='html-p'>The structure calculation of Kalata B1 was performed using the program “combined assignment and dynamics algorithm for NMR applications” (CYANA) [<a href="#B25-biomedicines-12-02216" class="html-bibr">25</a>]. The ensemble of representative structures was generated using a combination of NOESY cross-peaks, chemical shifts, and three-bond (<sup>3</sup>J<sub>HNα</sub>) coupling constants. In general, the structure calculation started with 1000 randomly generated structures. Two thousand and forty-eight randomly generated structures underwent simulated annealing using 16,384 molecular dynamics steps within the provided parameters. Two individual structure calculations were performed with additional parameters added on the next calculation as follows: (1) using torsion angle restraints generated based on the amino acid sequence and cis proline omega torsion angle for the P24 and (2) adding upper limit distance constraints between the protons generated from the NOE cross peak volume. When generating the upper limit distance constraints, an average upper distance limit of 4.2 Å was used. The ‘<span class='html-italic'>distance modify</span>’ command in CYANA was used to remove redundant and unnecessary distance constraints and apply the necessary pseudo-atom corrections. One hundred forty-nine upper limit distance constraints were generated and used for the structure calculation. The structure calculation selected the 20 lowest energy structures as the structural representation within the given parameters. The structure analysis was performed using PyMOL (the PyMOL Molecular Graphics System, Version 2.5.4, Schrodinger, LLC, New York, NY, USA).</div></section></section><section id='sec2dot4-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 2.4. Quantifying Cyclotides for Evaluation in Bioactivity Assays</h4><div class='html-p'>Freeze-dried samples of CyO3, CyO19, Kalata B1, Kalata B7, and Vitri E (~0.5–2.0 mg) were dissolved into 10% DMSO and quantified following the protocols described by Gerlach and colleagues [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>]. The concentrations of the peptides were determined spectrophotometrically using extinction coefficients determined by the ExPASy Protparam tool (<a href='https://web.expasy.org/protparam/' target='_blank' rel="noopener noreferrer">https://web.expasy.org/protparam/</a>, accessed on 14 August 2024) as follows: CyO3 (7365 mol<sup>−1</sup>cm<sup>−1</sup>), CyO19 (7365 mol<sup>−1</sup>cm<sup>−1</sup>), synthetic and natural Kalata B1 (5875 mol<sup>−1</sup>cm<sup>−1</sup>), Kalata B7 (7365 mol<sup>−1</sup>cm<sup>−1</sup>), and Vitri E (5875 mol<sup>−1</sup>cm<sup>−1</sup>). In addition to quantification via UV absorbance at 280 nm in a Nanodrop 2000 spectrophotometer (ThermoScientific), the peptides were also quantified as previously described by Gerlach et al. 2022 [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>], using Qubit 3.0 fluorometer (Thermo Fisher Scientific Invitrogen Life Technologies) protein quantification assays (Cat. No. Q33212).</div></section><section id='sec2dot5-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 2.5. Cell Culture</h4><div class='html-p'>SH-SY5Y human neuroblastoma cells (population doubling (PD) 20−24), T-98G, and U-87 MG human glioblastoma cells were removed from cryogenic storage, thawed rapidly in a water bath (37 °C) and reconstituted in 75 cm<sup>2</sup> flasks. For SH-SY5Y, the following media were used: Dulbecco’s modified Eagle’s medium/Nutrient F12 Ham (Sigma-Aldrich, Cat. No. D8437) containing 10% heat-inactivated fetal bovine serum, 4 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin. For U-87 MG and T-98G Eagle’s minimal essential medium (ATCC: The Global Bioresource Center, Cat. No. ATCC 30-2003), containing 10% heat-inactivated fetal bovine serum, 4 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin were used.</div><div class='html-p'>Cells were incubated at 37 °C in a humidified atmosphere of 5% CO2 and 95% air until recovered and ready for passaging (approximately 4 days). At between 80% and 90% confluence, the cells were trypsinized with warm (37 °C) sterile 0.5% Trypsin-EDTA (Thermo Fisher, Cat. No. 15400054) and passaged into 175 cm<sup>2</sup> flasks, where they were left to multiply until they reached 80−90% confluence. To plate cells: cultures were trypsinized, resuspended in 20 mL sterile PBS, counted using a cell counter (Olympus cell counter model R1), and plated in 96-well plates at an optimal density of 6000 cells/well in the designated culture media. The cells were left to adhere overnight before treatment the following day, when the culture medium was removed and replaced with the treatment medium. All incubations were conducted in triplicate wells and repeated at least three times (n = 3).</div></section><section id='sec2dot6-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 2.6. Phase Contrast Microscopy</h4><div class='html-p'>U-87 MG cells were plated into 96-well plates at 6000 cells/well, allowed to adhere overnight, then the medium changed to the following: (A) no treatment/control; (B) 150 µM temozolomide (TMZ); (C) synthetic Kalata B1 0.5 µM; and (D) 150 µM TMZ + 0.5 µM synthetic Kalata B1. The cultures were incubated for 72 h. The cultures were imaged on an Olympus IX83 inverted microscope using phase-contrast with an Olympus XM10 camera at 20× magnification (Tokyo, Japan). Images were acquired in cellSens Standard, then imported into Adobe Photoshop 23.4.2, where they were adjusted for exposure using auto “Levels”, sharpened using Topaz Laboratories Sharpen AI version 2.1.7 and cropped for consistency and to enable ease of comparison.</div></section><section id='sec2dot7-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 2.7. Determination of Half-Maximal Inhibitory Concentration (IC<sub>50</sub> Values) in TMZ and Cyclotides</h4><div class='html-p'>To calculate TMZ IC<sub>50</sub> values in each cell line, TMZ was dissolved in DMSO at a stock concentration of 103 mM, which was then diluted into culture medium on the day of use, with the DMSO final concentration always less than 0.1%. The following concentrations of TMZ were used in the experiments: 0.5, 5, 50, 500, 2500, 5000 μM. The media were sterilized for each treatment media with a 0.22 μm filter before they were added to cell cultures. To determine the IC<sub>50</sub> values for each cyclotide in each cell type, cyclotides were dissolved into 10% DMSO and then dilutions used to screen cyclotides with a final DMSO concentration of less than 0.1% The following concentrations of cyclotides were used: 0.1, 1, 5, 10 μM. For co-exposure experiments, a simultaneous exposure of cyclotide and TMZ at each respective dose was added to cell cultures (100 μL final volume per well).</div></section><section id='sec2dot8-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 2.8. MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] Assay</h4><div class='html-p'>In accordance with the methods developed by Mosmann and colleagues [<a href="#B26-biomedicines-12-02216" class="html-bibr">26</a>], the MTT assay was conducted using the Roche cell proliferation kit (Cat. No. 11465007001). Briefly, cells that are active can cleave MTT. The MTT is originally a yellow tetrazolium salt that undergoes a colorimetric change to purple formazan crystals, and the change in absorbance can be measured as an indicator of cell viability/proliferation. For these experiments, cells were plated into 96-well flat-bottom plates, as described in the cell culture above. After 24 h, the medium was replaced with 100 µL treatment medium consisting of either the control, TMZ, cyclotide alone, or a co-exposure treatment of TMZ + cyclotide. To align with our previous work [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>], experiments were conducted with a 72 h time frame for cytotoxicity evaluation.</div><div class='html-p'>The concentrations of cyclotides alone were as follows: 0, 0.1, 1, 5, and 10 µM of either CyO3, CyO19, Kalata B1, Kalata B7, and Vitri E. The maximum dose of 10 µM was used because previous research [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>] indicated this concentration provided sufficient cell death while preserving limited quantities of purified cyclotides. After a 72 h incubation period, plates were removed from the incubator and 10 µL of MTT was added to each well. The plates were returned to the incubator for an additional 4 h, after which 100 µL of a solubilization solution was added to each well. The plates were returned to incubation overnight, and the next day absorbances were read at 570 nm on a Molecular Devices FlexStation 3 using SoftMax Pro Microplate Data Acquisition and Analysis Software (version 7.0.2). All data were expressed as a percent of controls in a normalized response, and all incubations were conducted in triplicate wells (technical replicates) and in triplicate, independent incubations (n = 3).</div></section><section id='sec2dot9-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 2.9. IC<sub>50</sub> Value Calculations and Statistical Analysis</h4><div class='html-p'>We calculated the IC<sub>50</sub> values with a best fit method using nonlinear regression analysis with dose-dependent inhibition and an inhibitor versus normalized response. MTT assay results were analyzed statistically using one-way analysis of variance (ANOVA) and Tukey’s post hoc analysis, where <span class='html-italic'>p</span> < 0.05 was considered significant. All data were evaluated using GraphPad Prism for Mac OS (version 9.5.1). For each calculation, at least three independent experiments with 3 biological replicates for each experiment were performed.</div></section><section id='sec2dot10-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 2.10. Human Serum Stability Assay</h4><div class='html-p'>Established protocols [<a href="#B27-biomedicines-12-02216" class="html-bibr">27</a>,<a href="#B28-biomedicines-12-02216" class="html-bibr">28</a>] were used to analyze peptide stability in human serum isolated from male AB plasma (Sigma-Aldrich). Briefly, thawed plasma was centrifuged for 10 min at 15,000× <span class='html-italic'>g</span> to concentrate lipids. The supernatant was carefully removed with a Pasteur pipette and incubated for 15 min at 37 °C, 5% CO<sub>2</sub>. Stock synthetic Kalata B1 and TMZ (300 µM) were diluted 10 times with warmed plasma or warmed PBS for 0, 3, 8, and 24 h. Controls were synthetic Kalata B1 or TMZ in PBS that were incubated in parallel with human serum test compounds. Serum proteins were denatured with urea (final concentration of 3M) at 4 °C for 10 min., at each time point. Afterwards, trichloroacetic acid (7% final <span class='html-italic'>v</span>/<span class='html-italic'>v</span>) was added for 10 min to precipitate the proteins. Each sample was centrifuged for 10 min at 15,000× <span class='html-italic'>g</span>; the supernatant was removed an evaluated in triplicate using UPLC-MS.</div></section></section><section id='sec3-biomedicines-12-02216' type='results'><h2 data-nested='1'> 3. Results</h2><section id='sec3dot1-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 3.1. Verification of Synthetic Kalata B1</h4><section id='sec3dot1dot1-biomedicines-12-02216' type=''><h4 class='' data-nested='3'> 3.1.1. UPLC-MS Synthetic Kalata B1 Verification</h4><div class='html-p'>In previous research [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>], we demonstrated that our stock of purified natural Kalata B1 was 95.4% pure. We commissioned CSBio (Menlo Park, CA, USA) to synthesize 30 mg of synthetic Kalata B1, which is documented at 97.51% pure. They synthesized a linear cyclotide, cyclized it, and folded it using head-to-tail lactam bonding and natural oxidation (<a href="#app1-biomedicines-12-02216" class="html-app">Figure S1</a>). The present UPLC-MS analyses of both synthetic and natural Kalata B1 indicated they had an elution time of 5.05–5.25 min., and both have a <span class='html-italic'>m</span>/<span class='html-italic'>z</span> of 964.77. There was a slight shift in the elution time, which was attributed to matrix interference (<a href="#biomedicines-12-02216-f002" class="html-fig">Figure 2</a>).</div></section><section id='sec3dot1dot2-biomedicines-12-02216' type=''><h4 class='' data-nested='3'> 3.1.2. Orbitrap Analysis of Exploris 480 Mass Spectrometry–Liquid Chromatography</h4><div class='html-p'>The Orbitrap analysis of the alkylated and enzymatically digested peptides revealed the presence of ions consistent with digested Kalata B1 (e.g., <a href="#biomedicines-12-02216-f003" class="html-fig">Figure 3</a>). The assessment of the masses observed showed good consistency with the exact masses expected, with <span class='html-italic'>m</span>/<span class='html-italic'>z</span> values of 845.3799 and 1215.9891 observed for peptides 1 and 2, respectively. Sequence analysis of the two peptides showed daughter ions consistent with the expected sequences (<a href="#biomedicines-12-02216-t001" class="html-table">Table 1</a> and <a href="#biomedicines-12-02216-t002" class="html-table">Table 2</a>). For peptide 1, all the amino acids could be found in this 8-mer peptide (<a href="#biomedicines-12-02216-t001" class="html-table">Table 1</a>), whereas only 19 of the 21 amino acids could be sequenced for peptide 2 (<a href="#biomedicines-12-02216-t002" class="html-table">Table 2</a>). However, given the expected vs. observed <span class='html-italic'>m</span>/<span class='html-italic'>z</span> ratios of the parent peptides (845.3821 vs. 845.3799, peptide 1 and 1215.9894 vs. 1215.9891, peptide 2), both peptides match Kalata B1, with 93% coverage of the peptide according to Proteome Discoverer (Thermo Scientific) assessment of the sequence.</div></section><section id='sec3dot1dot3-biomedicines-12-02216' type=''><h4 class='' data-nested='3'> 3.1.3. The Structural Characteristics of Kalata B1 Using NMR Spectroscopy</h4><div class='html-p'>The DQFC, TOCSY, and NOESY spectra were used for the sequence-specific backbone chemical shift assignment of Kalata B1 (<a href="#biomedicines-12-02216-f004" class="html-fig">Figure 4</a>A). A representative three-dimensional structure determined by NMR spectroscopy is also shown (<a href="#biomedicines-12-02216-f004" class="html-fig">Figure 4</a>B). The six cysteine residues (I to VI) and the corresponding disulfide bonds in the three-dimensional fold of the synthetic Kalata B1 are indicated. All the proton chemical shifts of Kalata B1 were assigned (<a href="#biomedicines-12-02216-t003" class="html-table">Table 3</a>), except for the protons on the end of the guanidino group from Arg 28, the thiol protons from the six cysteine residues, and the protons from the carboxamide group of Asn 15 and Asn 29. <a href="#biomedicines-12-02216-t001" class="html-table">Table 1</a> also lists the three-bond <sup>1</sup>H-<sup>1</sup>H coupling constant (<sup>3</sup>J<sub>NH-CαH</sub>).</div><div class='html-p'>After assigning the backbone chemical shift and confirming the cyclic peptide nature of Kalata B1 via the NOE cross peak between H<sup>α</sup> of Asn 29 to the H<sup>N</sup> of Gly 1 (<a href="#app1-biomedicines-12-02216" class="html-app">Figure S2b</a>), the DQFC and TOCSY were used to complete the chemical shift assignment of the side chains. Kalata B1 has three proline residues at positions 3, 17, and 24. Proline can exist as a cis or trans isomer due to the peptide bond linkage, which can be identified via NMR [<a href="#B29-biomedicines-12-02216" class="html-bibr">29</a>]. Pro 3 and Pro 17 were present as trans isomers, as indicated by the NOE cross peak between the H<sup>δ</sup> of the proline to the H<sup>α</sup> of the preceding residue on its N-terminus side (<a href="#app1-biomedicines-12-02216" class="html-app">Figure S2c</a>). On the other hand, pro 24 was present as a cis isomer, as evidenced by the NOE cross peak between the Hα of the proline and the H<sup>α</sup> of the preceding residue on its N-terminus side (see <a href="#app1-biomedicines-12-02216" class="html-app">Figure S2c</a>). Initial structures were generated without the NOE distance constraints to convey the cyclic nature of Kalata B1 via the NMR data. An evolution of the structure generation process is illustrated (<a href="#app1-biomedicines-12-02216" class="html-app">Figure S3</a>), showing the structure before and after adding the NOE distance constraints.</div><div class='html-p'>The 20 Kalata B1 structures generated by CYANA using NOE distance constraints have no distance violation greater than 0.3 Å (<a href="#app1-biomedicines-12-02216" class="html-app">Figure S4a</a>). The average backbone RMSD to mean was 0.56 ± 0.25 Å (0.27…1.53 Å; 20 structures), and the average heavy atom RMSD to mean was: 0.87 ± 0.29 Å (0.59…2.02 Å; 20 structures). A three-dimensional alignment of the structure of Kalata B1 presented here has an RMSD of 1.18 Å, with reference to the solution structure of Kalata B1 from the natural resources (PDB 2khb).</div></section></section><section id='sec3dot2-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 3.2. Determination of IC<sub>50</sub> Values in TMZ and Cyclotides</h4><div class='html-p'>The IC<sub>50</sub> values of natural and synthetic cyclotides, as well as TMZ, are provided in <a href="#biomedicines-12-02216-t004" class="html-table">Table 4</a>. As illustrated, each cyclotide (CyO3, CyO19, natural Kalata B1, synthetic Kalata B1, and Vitri E) showed much lower IC<sub>50</sub> values compared to the TMZ IC<sub>50</sub> values. Synthetic Kalata B1 was approximately 2-fold more potent in the T-98G cells compared to the U-87 MG cells. Natural Kalata B1 (IC<sub>50</sub> = 7.9 µM) was comparable to the synthetic Kalata B1 (IC<sub>50</sub> value = 5.4 µM) in U-87 MG cells. The IC<sub>50</sub> values were calculated using nonlinear regression analysis with dose-dependent inhibition and an inhibitor vs. normalized response, as calculated by GraphPad Prism 9.5.1. For details of cell culture, see Materials and Methods. <sup>a</sup> represents data previously published [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>].</div><div class='html-p'>The IC<sub>50</sub> values of TMZ, natural cyclotides (CyO3, CyO19, Kalata B1, Kalata B7, and Vitri E), and the synthetic cyclotide Kalata B1 were evaluated in T-98G human glioblastoma, U-87 MG human glioblastoma, and SH-SY5Y human neuroblastoma cell cultures using MTT assays by plating cells in 96-wells with various concentrations of cyclotides and a 72 h incubation period. The IC<sub>50</sub> value can be defined as the concentration of the drug (cyclotide), whereby 50% of the cells survive, and cell viability was then expressed as a percentage of control (cells incubated with DMSO < 0.01%). As illustrated in <a href="#biomedicines-12-02216-f005" class="html-fig">Figure 5</a>, lower concentrations of TMZ were required to obtain an IC<sub>50</sub> value in T-98G cells (1160.0 µM) compared to U-87 MG cells (1617.0 µM).</div><div class='html-p'>Increasing concentrations of plant-derived cyclotides (CyO3, CyO19, Kalata B1, Kalata B7, and Vitri E) were evaluated in MTT assays to determine the IC<sub>50</sub> values in U-87 MG glioblastoma cell cultures. The cyclotides exhibited dose-dependent cytotoxicity, and the Best Fit method for IC<sub>50</sub> value was calculated with nonlinear regression analysis using dose-dependent inhibition with an inhibitor vs. normalized response and confidence intervals of 95%, as calculated by GraphPad Prism 9.0 for iOS. Cyclotides exhibited a dose-dependent cytotoxicity, with IC<sub>50</sub> values ranging from 2.4 to 21.1 µM. The cyclotides demonstrating the most to least potent cytotoxicity were as follows: CyO19 (IC<sub>50</sub> value = 2.4 µM), CyO3 (IC<sub>50</sub> value = 2.6 µM), Kalata B1 (IC<sub>50</sub> value = 7.9 µM), Kalata B7 (IC<sub>50</sub> value = 10.1 µM), and Vitri E (IC<sub>50</sub> value = 21.1 µM) (<a href="#biomedicines-12-02216-f006" class="html-fig">Figure 6</a>).</div><div class='html-p'>To evaluate the cytotoxicity of plant-derived Kalata B1 and synthetic Kalata B1, MTT assays were performed with T-98G, U-87 MG, and SH-SY5Y cell cultures. The concentrations (0–10 µM) were evaluated during a 72 h incubation period, with Best Fit IC<sub>50</sub> values calculated as previously described, which indicated that each cyclotide had an IC<sub>50</sub> value of <22 µM for the cell lines investigated. The natural Kalata B1 had an IC<sub>50</sub> value of 5.6 µM in the SH-SY5Y cells and an IC<sub>50</sub> value of 7.9 µM in the U-87 MG cell cultures. The IC<sub>50</sub> values of synthetic Kalata B1 were 2.6 µM in T-98G cells and 5.4 µM in U-87 MG cells. Thus, the activity of natural Kalata B1 was comparable to synthetic Kalata B1 (<a href="#biomedicines-12-02216-f007" class="html-fig">Figure 7</a>).</div></section><section id='sec3dot3-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 3.3. Cytotoxicity of Co-Exposure with Cyclotides and TMZ in U-87 MG and T-98G Cells</h4><div class='html-p'>In the U-87 MG glioblastoma cells, the experimentally determined IC<sub>50</sub> Best Fit value for TMZ was 1617.0 µM (<a href="#biomedicines-12-02216-f005" class="html-fig">Figure 5</a>), and the IC<sub>50</sub> Best Fit value for synthetic Kalata B1 was 5.4 µM (<a href="#biomedicines-12-02216-f007" class="html-fig">Figure 7</a>). We were particularly interested in improving the sensitivity of glioblastoma cells to TMZ; therefore, we incubated cultures with several reduced concentrations of TMZ and synthetic Kalata B1, alone and in combination. MTT assays (as previously described) were employed to evaluate cell viability following co-exposure treatments. Cells were incubated with Kalata B1 (K) (0.5, 1.0, 2.0, and 4.0 µM) alone, TMZ (T) (100, 200, 400, and 800 µM) alone, and then co-exposed to synthetic Kalata B1 and TMZ.</div><div class='html-p'>The results of the one-way analysis of variance (ANOVA), followed by Tukey’s multiple comparisons, were graphed in GraphPad Prism version 9 for iOS and indicate that there was a significant reduction in cell viability in the following co-exposure treatments: (K 0.5 µM + T 100 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>A), (K 0.5 µM + T 400 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>H), (K 0.5 + T 800 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>K), (K µM 1 + T 100 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>B), (K 1 µM + T 200 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>F), (K 1 µM + T 400 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>I), (K 1 µM + T 800 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>L), (K 2 µM + T 100 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>C), (K 2 µM + T 200 µM; Figurer 8F), (K 2 µM + T 400 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>J), (K 2 µM + T 800 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>M), (K 4 µM + T 100 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>A), (K 4 µM + T 200 µM; <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>G). Although 3 out of the 16 co-exposure treatments analyzed (K 0.5 µM + T 200 µM, K 4 µM + T 400 µM, K µM + T 800 µM) did not significantly increase TMZ-induced cytotoxicity (<a href="#app1-biomedicines-12-02216" class="html-app">Figure S5</a>), the results confirm that 81.3% of the combination treatments analyzed provide support that synthetic Kalata B1 can chemosensitize cells to TMZ, and TMZ concentrations over 16-fold lower than the IC<sub>50</sub> Best Fit value for TMZ (1615.1 µM, <a href="#biomedicines-12-02216-f005" class="html-fig">Figure 5</a>B) can still result in significant cell death in U-87 MG glioblastoma cells (<a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>). The methodology used to analyze co-exposure data are similar to a previous publication [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>], and the results are consistent with co-exposure treatments of other naturally occurring cyclotides, TMZ, and glioblastoma [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>].</div><div class='html-p'>In the T-98G glioblastoma cells, the experimentally determined IC<sub>50</sub> Best Fit value for TMZ was 1160.0 µM (<a href="#biomedicines-12-02216-f005" class="html-fig">Figure 5</a>), and the IC<sub>50</sub> Best Fit value for synthetic Kalata B1 was 2.6 µM (<a href="#biomedicines-12-02216-f007" class="html-fig">Figure 7</a>). To address our interest in chemosensitizing glioblastoma cells to TMZ, we used previously described MTT assays to analyze cell viability with T-98G cells exposed to reduced concentrations of TMZ and synthetic Kalata B1. Cells were incubated with synthetic Kalata B1 (K) (0.25, 0.5, 1.0, and 2.0 µM) alone, TMZ (T) (75, 150, 300, and 600 µM) alone, and then co-exposed to Kalata B1 and TMZ.</div><div class='html-p'>Results of one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparisons were graphed in GraphPad Prism version 9 for iOS and indicate that there was a significant reduction in cell viability in the following co-exposure treatments: (K 0.25 + T 75 µM; <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>A), (K 0.25 + T 150 µM; <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>E), (K 0.5 + T 75 µM; <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>B), (K 0.5 + T 150 µM; <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>C), (K 0.5 + T 300 µM; <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>G), (K 0.5 + T 600 µM; <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>H), (K1 + T 150 µM; <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>D), (K 1 + T 300 µM; <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>F), and (K 2 + T 600 µM; <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>I). Although some treatments (K 1 µM + T 75 µM, K 22 µM + T 75 µM, K 2 µM + T 150 µM, K 0.25 + T 300 µM, K 2 µM + T 300 µM, K 1 µM + T 600 µM) did not cause significant differences in co-exposure treatments (<a href="#app1-biomedicines-12-02216" class="html-app">Figure S6</a>), at least 56.2% of the co-treatments with synthetic Kalata B1 chemosensitized T-98G glioblastoma cells to TMZ. Furthermore, the lowest co-exposure concentration (K 0.25 µM + T 75 µM; <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>A) used a concentration of TMZ 15-fold lower that the IC<sub>50</sub> Best Fit value for TMZ in T-98G cells (1167.6 µM; <a href="#biomedicines-12-02216-f005" class="html-fig">Figure 5</a>A) and still caused significantly greater cell death compared to synthetic Kalata B1 alone (0.25 µM) or TMZ alone (75 µM) (<a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>). Low concentrations of Kalata B1, (0.5 μM in U-87 MG cells and 0.25 μM in T-98G cells) could produce a stimulatory effect, as illustrated in <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a> and <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>, when analyzed as a percent of control (no treatment). This activity has been documented in other cancer cell lines, such as human malignant glioblastoma tumor cells (U-251) and primary human umbilical vein endothelial cells (HUVEC) [<a href="#B30-biomedicines-12-02216" class="html-bibr">30</a>]. The analysis methods used were aligned with a previous publication [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>], and results are consistent with the indication that cyclotides at various concentrations may increase TMZ toxicity in glioblastoma cell lines.</div></section><section id='sec3dot4-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 3.4. Phase Contrast Microscopy to Visualize Cytotoxicity in U-87 MG Cells</h4><div class='html-p'>U-87 MG cells play a significant role in the development of glioblastoma multiforme, and they develop resistance rapidly; therefore, we are interested in determining if differences could be seen in untreated (control) U-87 MG glioblastoma cells and those treated with TMZ (150 µM), synthetic Kalata B1 (0.5 µM), and co-treatments (150 µM TMZ + 0.5 µM synthetic Kalata B1). We performed phase contrast microscopy to visualize the changes induced by treatments that previously showed significant results in cell viability assays. In cultures incubated with 150 µM TMZ (<a href="#biomedicines-12-02216-f010" class="html-fig">Figure 10</a>B) and 0.5 µM synthetic Kalata B1 (<a href="#biomedicines-12-02216-f010" class="html-fig">Figure 10</a>C), little differences were observable between the control cultures (<a href="#biomedicines-12-02216-f010" class="html-fig">Figure 10</a>A) and the respective treatment. This corresponded with the MTT cell viability assays, which returned almost no cell death for the U-87 MG cells treated with 0.5 µM synthetic Kalata B1, and less than 15% cell death for cells treated with ≤200 µM TMZ. Conversely, in cells undergoing co-exposure (150 µM TMZ + 0.5 µM synthetic Kalata B1), noticeable changes in microscopy images correlated with finding of cell death in <a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>, with various shrunken, granular, blebbing cells, which suggests apoptosis (<a href="#biomedicines-12-02216-f010" class="html-fig">Figure 10</a>D). Conversely, microscopy imaging of T-98G glioblastoma cells was not as clear, possibly due to variation in the pattern of growth (<a href="#app1-biomedicines-12-02216" class="html-app">Figure S7</a>).</div></section><section id='sec3dot5-biomedicines-12-02216' type=''><h4 class='html-italic' data-nested='2'> 3.5. Synthetic Kalata B1 Remains Stable in Human Serum</h4><div class='html-p'>We conducted peptide stability assays in human serum isolated from male AB plasma and then analyzed with UPLC-MS. Synthetic Kalata B1 and TMZ were incubated with human serum and the portion of the peptide remaining was quantified by calculating the area of the respective elution peak in UPLC-MS, compared to the control. <a href="#biomedicines-12-02216-f011" class="html-fig">Figure 11</a> indicates that at data points of 3 h, 8 h, and 24 h, synthetic Kalata B1 remains stable, whereas the concentration of TMZ decreases to less than 50% at 3 h, less than 30% at 8 h, and <5% at 24 h.</div></section></section><section id='sec4-biomedicines-12-02216' type='discussion'><h2 data-nested='1'> 4. Discussion</h2><div class='html-p'>Glioblastoma multiforme (GBM) affects both children and adults and is the most aggressive type of cancer in the brain. Post-diagnosis, patients often survive fewer than 16 months. It is estimated that less than 7% of patients diagnosed with GBM survive longer than five years after diagnosis, in part, because resection and radiation alone are not efficacious, and many patients become resistant to currently approved FDA drugs [<a href="#B31-biomedicines-12-02216" class="html-bibr">31</a>,<a href="#B32-biomedicines-12-02216" class="html-bibr">32</a>]. In this study, we investigated the effects of cyclotides on the U-87 MG and T-98G glioblastoma cell lines to determine if they can chemosensitize glioblastoma cell lines to the FDA-approved drug temozolomide (TMZ). TMZ does offer some hope to patients as a chemotherapeutic agent; however, roughly 50% of patients do not respond to TMZ, and those patients in which TMZ is initially efficacious often develop rapid drug resistance [<a href="#B33-biomedicines-12-02216" class="html-bibr">33</a>]. Therefore, studies to discover adjuvant methods to treat GBM are vital.</div><div class='html-p'>To this end, we investigated several naturally occurring cyclotides (CyO3, CyO19, Kalata B1, Kalata B7, Vitri E) and one synthetic cyclotide, Kalata B1. The plant-derived compounds are commonly found in <span class='html-italic'>Viola odorata</span> L. and <span class='html-italic'>Viola tricolor</span> L., two species of violets in the family Violaceae. Although the cytotoxicity of Kalata B1 has been reported in U-87 and U-251 glioblastoma cells using an MTT assay [<a href="#B30-biomedicines-12-02216" class="html-bibr">30</a>], different experimental conditions, incubation periods, and dyes for assessing cell viability were used in the current study, yet the results were still comparable. Tang and colleagues [<a href="#B34-biomedicines-12-02216" class="html-bibr">34</a>] evaluated the cell viability of several cyclotides in a sulforhodamine B (SRB) assay, and several of them such as Cycloviolacin O2, Vitri A, and Vitri F displayed toxicity towards U-251 glioblastoma cells that was equipotent to that of the U-87 MG and T-98G cellular data provided in the current research (<a href="#biomedicines-12-02216-t004" class="html-table">Table 4</a>).</div><div class='html-p'>Cyclotides isolated from <span class='html-italic'>Viola sumatrana</span> Miq. also lead to cytotoxicity in MTT assays, with CyO2 displaying more potent activity (IC<sub>50</sub> = 0.45 and 0.79 µM) compared to Kalata B1 (IC<sub>50</sub> = 3.21 and 10.88 µM) in the U-87 and U-251 glioblastoma cells, respectively [<a href="#B35-biomedicines-12-02216" class="html-bibr">35</a>]. Our previous research also indicates that cyclotides such as CyO2 (IC<sub>50</sub> = 4.5 µM), CyO13 (IC<sub>50</sub> = 2.57 µM), and Varv peptide A (IC<sub>50</sub> = 2.91 µM) display cytotoxicity in a range similar to the data presented in this research (IC<sub>50</sub> = 2.4–21.1 µM) (<a href="#biomedicines-12-02216-f006" class="html-fig">Figure 6</a>). This indicates that cyclotides alone may be promising compounds for further investigation in GBM chemotherapy. The five cyclotides in this research displayed dose-dependent cytotoxicity in human neuroblastoma (SH-SY5Y) and human glioblastoma (U-87 MG, and T-98G) cells (<a href="#biomedicines-12-02216-t004" class="html-table">Table 4</a>). Previous studies indicate that the cytotoxic potency of cyclotides can be significantly affected by substitutions or removal of amino acids in the cyclotide sequences. The most striking evidence of this fact was a 48-fold decrease in cytotoxicity in lymphoma cells that Lindholm and colleagues [<a href="#B36-biomedicines-12-02216" class="html-bibr">36</a>] observed when the conserved glutamic acid (E) was removed from cyclotides. The extent of surface exposure of hydrophobic patches in cyclotides has been known to affect bioactivity, which indicates that structure plays an important role in bioactivity. In the present research, two bracelet cyclotides (CyO3 and CyO19) and three Möbius cyclotides (Kalata B1, Kalata B7, and Vitri E) were screened. CyO3 and CyO19 have varying amino acids in loop 3 and loop 6, while Kalata B1, Kalata B7, and Vitri E have a distinct complement of amino acids in loops 3, 5, and 6. Despite these differences, all the cyclotides in this research displayed IC<sub>50</sub> values of <22 µM.</div><div class='html-p'>Challenges commonly associated with treating cancer can include low drug uptake across the cellular lipid bilayer, the activity of pumps, such as p-glycoprotein, that efflux drugs out of the cell, and the impenetrability of the blood–brain barrier [<a href="#B11-biomedicines-12-02216" class="html-bibr">11</a>,<a href="#B37-biomedicines-12-02216" class="html-bibr">37</a>]. Our previous research provided supporting evidence that cyclotides such as CyO2 can act as adjuvants in drug-resistant breast cancer cells, whereby cyclotides sensitize human breast adenocarcinoma (MCF-7) and drug-resistant human breast adenocarcinoma (MCF-7/ADR) cells to the FDA-approved anti-cancer drug, doxorubicin. In that case, one mechanism of cytotoxicity was most likely the development of tetramer pores on the plasma membrane of the cancer cell, which allowed for the passage and uptake of doxorubicin using the SYTOX green assay. Interestingly, Cycloviolacin O2 did not produce significant membrane disruption in primary human brain endothelial cells [<a href="#B11-biomedicines-12-02216" class="html-bibr">11</a>]. The literature indicates that the bioactivity of Kalata B1 has yet to be determined in noncancerous human brain cells; therefore, another goal of our future research will include an evaluation of Kalata B1 in unaffected brain cells.</div><div class='html-p'>Although a mechanistic evaluation of Kalata B1 bioactivity was beyond the scope of this research, a brief discussion of the previous mechanistic studies is justified. For instance, Henriques and colleagues [<a href="#B35-biomedicines-12-02216" class="html-bibr">35</a>] provided supporting evidence that cyclotides can target phosphatidylethanolamine (PE)-enriched phospholipids and penetrate the cell via both endocytosis and membrane translocation. An additional study indicated that at least one mechanism of activity of Kalata B1 and Kalata B2 is consistent with the formation of lipidic toroidal pores that decrease the critical packing parameter and act much like a surfactant [<a href="#B38-biomedicines-12-02216" class="html-bibr">38</a>]. Furthermore, bioactivity studies for immune-related disorders have investigated additional mechanisms of bioactivity, with investigations documenting that a Kalata B1 mutant, named [T20k] Kalata B1, could decrease the expression of interleukin-2 (IL-2), IL-2 cytokine secretion, and IL-2 gene expression [<a href="#B39-biomedicines-12-02216" class="html-bibr">39</a>]. Since these reports indicate the mechanisms of cyclotide bioactivity are complex, future evaluations with synthetic Kalata B1 and any analogs should also focus on delineating their specific mechanisms of action regarding GBM.</div><div class='html-p'>Here, one aim of our experiments was to assess if co-exposure of TMZ with synthetic Kalata B1 could increase the sensitivity of U-87 MG and T-98G cells to TMZ-induced cytotoxicity. The results lay the foundation for transitioning towards an in vivo xenograft mouse model to begin determining whether a combination approach with cyclotides and TMZ may reduce the required effective dose of TMZ and perhaps decrease the development of TMZ resistance in glioblastoma cells. To be considered potentially efficacious, the combination treatment must be significantly different from that of synthetic Kalata B1 alone or TMZ alone, which was indicated in the co-exposure treatment in U-87 MG (<a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a>) and T-98G (<a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>) glioblastoma cell lines. Although obtaining the IC<sub>50</sub> values of combination treatments was not a primary objective of this research, future studies will focus on obtaining full dose response curves for each agent (synthetic Kalata B1 and TMZ), as well as fixed ratio (synthetic Kalata B1 + TMZ) treatments followed by an analysis of the combination index to determine if the activity is synergistic, additive, or antagonistic.</div><div class='html-p'>The development of an in vivo mouse model is vital, yet it requires expertise, significant finances, and large quantities of the target cyclotide. To this end, an early investigation with the cyclotide Cycloviolacin O2 was completed, demonstrating that it had an abrupt in vivo toxicity profile, since a single injection at 2 mg/kg was lethal, yet there were no signs of discomfort in mice treated with 1.5 mg/kg in tumor mouse models [<a href="#B40-biomedicines-12-02216" class="html-bibr">40</a>]. A mouse model of anaplastic large cell lymphoma (ALCL) was developed, and the Kalata B1 mutant, named [T20K] Kalata B1, decreased tumor weight and increased apoptosis, which led researchers to assert that cyclotide-based drugs may be a promising therapy for patients with ALCL [<a href="#B41-biomedicines-12-02216" class="html-bibr">41</a>]. A full report on the safety and efficacy of synthetic Kalata B1 has not been conducted in mouse models of GBM; therefore, one of our future objectives is the development of an in vivo GBM mouse model.</div><div class='html-p'>Our previous research indicates that cyclotides (CyO2 and Varv peptide A) chemosensitize glioblastoma cells to TMZ. In fact, the data indicated that 8-fold less TMZ was required to cause significant cell death in U-87 MG cells exposed to CyO2 co-exposure with TMZ. Likewise, 2-fold less TMZ was needed for significant cytotoxicity in U-87 MG cells co-exposed to Varv peptide A. The data presented here indicate that the chemosensitizing effects of synthetic Kalata B1 are comparable to the activity illustrated with CyO2 and Varv peptide A [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>]. Therefore, we provide supporting evidence that synthetic Kalata B1 increased TMZ-induced cell death in both U-87 MG and T-98G glioblastoma cells (<a href="#biomedicines-12-02216-f008" class="html-fig">Figure 8</a> and <a href="#biomedicines-12-02216-f009" class="html-fig">Figure 9</a>). Although MTT assays provide valuable information regarding cell viability, it is important to fully evaluate the effects of synthetic Kalata B1 on TMZ for GBM treatment; therefore, future priorities include investigating the cyclotide’s activity in other bioactivity assays, such as the lactate dehydrogenase (LDH) assay, the colony formation assay, and the blood–brain barrier assay.</div><div class='html-p'>Symptoms of GBM include seizures, headaches, nausea, and memory loss, and the side effects of TMZ can not only intensify headaches and nausea but also include diarrhea, constipation, loss of appetite, and sores in the mouth and throat. TMZ is an oral DNA alkylating agent that induces cell cycle arrest at G2/M, and its cytotoxicity is mediated by the addition of methyl groups at N<sup>7</sup> and O<sup>6</sup> sites on guanines and the O<sup>3</sup> site on adenines in genomic DNA. A thymine instead of a cytosine is inserted during the alkylation of the O<sup>6</sup> site on guanine, which can cause cell death during DNA replication. Patient resistance to TMZ is a significant challenge in TMZ chemotherapy. The mechanism of TMZ resistance involves methylated sites that can remain mutated, be fixed via DNA mismatch repair (MMR), be dealkylated by the action of a demethylating enzyme including O<sup>6</sup>-methylguanine methyltransferase (MGMT), or be removed by base excision repair (BER) by the activity of DNA glycosylase, such as alkylpurine-DNA-N-glycosylase (APNG). When MMR is expressed, human glioblastoma cells are TMZ-sensitive, and when MGMT, BER, and APNG proteins are expressed, then the cells develop resistance to TMZ [<a href="#B31-biomedicines-12-02216" class="html-bibr">31</a>]. In the current research, the IC<sub>50</sub> value of the TMZ-resistant cell line T-98G (1160.0 μM) is within the range of the IC<sub>50</sub> values (>250–1585 μM) summarized by Lee [<a href="#B31-biomedicines-12-02216" class="html-bibr">31</a>]. Interestingly, the IC<sub>50</sub> value of the report TMZ-sensitive U-87 MG cell line (1617.0 μM) is higher than the range (7 to less than 500 μM) reviewed. Human glioblastoma cells can acquire resistance rapidly or over time; therefore, a speculative explanation for the higher IC<sub>50</sub> value of the U-87 MG cell line in this study is that these cells may have been expressing, singularly or in combination, MGMT, BER, and APNG proteins.</div><div class='html-p'>One of our objectives is to promote research that investigates methods to decrease the required concentrations of drugs such as TMZ for efficacy in vivo. Therefore, it is important to analyze the stability of cyclotides in human serum. The trypsin inhibitor cyclotide, MCoTI-II, has been extracted from the seeds of <span class='html-italic'>Momordica cochinchinensis</span> Spreng. [Cucurbitaceae]. Chan and colleagues [<a href="#B27-biomedicines-12-02216" class="html-bibr">27</a>] performed serum stability assays and documented that MCoTI-II remained stable in human serum for 24 h with 100% peptide remaining. Huang and colleagues [<a href="#B28-biomedicines-12-02216" class="html-bibr">28</a>] repeated these experiments with MCoTI-II and an analog, MCo-RMI. Both peptides remained stable in human serum for at least 24 h. We adopted their methodology and herein provide evidence that synthetic Kalata B1 also remains stable in human serum isolated from male AB plasma for at least 24 h.</div><div class='html-p'><span class='html-italic'>Momordica cochinchinensis</span> is a perennial melon that expresses the trypsin inhibitor MCoTI-II, and it is a cell-penetrating peptide, which is a group of peptides reported to cross the blood–brain barrier. Wang and colleagues [<a href="#B42-biomedicines-12-02216" class="html-bibr">42</a>] compared the four-disulfide peptide (chlorotoxin) to the three-disulfide peptide (MCoTI-II) and demonstrated that chlorotoxin did cross the blood–brain barrier in a mouse model, yet MCoTI-II was not translocated across the brain. Another report of a radio-labelled analog modified from another trypsin-inhibitor cyclotide, MCoTI-I, extracted from <span class='html-italic'>M. cochinchinensis</span>, indicated that the analog ([<sup>64</sup>CU] MCo-CVX-6D) increased the uptake of U-87-stb-C-X-C chemokine receptor type 4 (CXCR4)-expressing cells in vitro and in vivo [<a href="#B43-biomedicines-12-02216" class="html-bibr">43</a>]. MCoTI-I and MCoTI-II are characterized as cyclotides in the trypsin-inhibitor subfamily; however, they have also been regarded as cyclic knottins because they contain a cystine knot motif as with all other cyclotides, but they have no further sequence similarity with the bracelet and Möbius subfamilies of cyclotides [<a href="#B43-biomedicines-12-02216" class="html-bibr">43</a>,<a href="#B44-biomedicines-12-02216" class="html-bibr">44</a>,<a href="#B45-biomedicines-12-02216" class="html-bibr">45</a>]. Therefore, future directives include optimizing a PAMPA blood–brain barrier (BBB) model to quantify the passage of synthetic Kalata B1 (a cyclotide in the Möbius subfamily) across a simulated BBB and developing a xenograft mouse model of GBM to study cyclotide bioactivity in vivo.</div></section><section id='sec5-biomedicines-12-02216' type='conclusions'><h2 data-nested='1'> 5. Conclusions</h2><div class='html-p'>In conclusion, this research identified cyclotides with techniques including UPLC-PDA and HPLC-UV and provided sequence confirmation for synthetic Kalata B1 using orbitrap LC-MS, followed by structural confirmation with NMR spectrometry. Human stability assays verified that synthetic Kalata B1 remains stable in serum for at least 24 h, and the evaluation of bioactivity in human glioblastoma cell lines (U-87 MG and T-98G) indicate that the cyclotides, Cycloviolacin O3, Cycloviolacin O19, natural Kalata B1, synthetic Kalata B1, and Vitri E cause dose-dependent cytotoxicity. Co-exposure experiments with synthetic Kalata B1 and TMZ provided initial proof-of-concept that the synthetic Kalata B1 may enhance TMZ toxicity in U-87 MG and T-98G glioblastoma cells. Therefore, continued research on the development of an adjuvant method towards the treatment of GBM with cyclotides and TMZ is warranted.</div></section> </div> <div class="html-back"> <section><section id='app1-biomedicines-12-02216' type=''><h2 data-nested='1'> Supplementary Materials</h2><div class='html-p'>The following supporting information can be downloaded at: <a href='https://www.mdpi.com/article/10.3390/biomedicines12102216/s1' target='_blank' rel="noopener noreferrer">https://www.mdpi.com/article/10.3390/biomedicines12102216/s1</a>, Figure S1: Verification of synthetic Kalata B1 purity from CSBio (Menlo Park, CA, USA); Figure S2: The structural feature of Kalata B1; Figure S3: The folding of Kalata B1 from a random coil structure to cyclic structure using NOE distance constraints. Figure S4: Synthetic and natural Kalata B1; Figure S5: synthetic Kalata B1 did not significantly increase TMZ-induced cytotoxicity in U-87 MG glioblastoma cell culture when using the following co-exposure concentrations: Kalata B1 (K) 0.5 µM + Temozolomide (T) 200 µM, K 4 µM + T 400 µM, and K 4 µM + T 800 µM; Figure S6: Synthetic Kalata B1 did not significantly increase TMZ-induced cytotoxicity in T-98G glioblastoma cell culture when using the following co-exposure concentrations: Kalata B1 (K) 1 µM + Temozolomide 75 µM, K 2 µM + T 75 µM, K 2 µM + T 150 µM, K 0.25 + T 300 µM, K 2 µM + T 300 µM, K 1 µM + T 600 µM; Figure S7: Light microscopy of T-98G cultures treated with Kalata B1 and temozolomide.</div></section></section><section class='html-notes'><h2 >Author Contributions</h2><div class='html-p'>Conceptualization, S.L.G., J.S.M., R.A.D., S.A.B. and P.A.C.; formal analysis, S.L.G., J.S.M., R.A.D., S.A.B., C.H. and V.V.K.; funding acquisition, V.V.K., U.G. and P.A.C.; methodology, S.L.G., J.S.M., R.A.D., S.A.B., C.H., V.V.K., U.G., S.G., B.S. and P.A.C.; resources, V.V.K., U.G. and P.A.C.; software, S.L.G., J.S.M., R.A.D., C.H. and V.V.K.; supervision, S.A.B., V.V.K., U.G. and P.A.C.; writing—original draft, S.L.G., J.S.M., R.A.D., S.A.B. and V.V.K.; writing—review and editing, S.L.G., J.S.M., R.A.D., S.A.B., V.V.K., U.G., S.G., B.S. and P.A.C. All authors have read and agreed to the published version of the manuscript.</div></section><section class='html-notes'><h2>Funding</h2><div class='html-p'>The 600 MHz spectrometer (VVK California State University Fresno) was funded by NSF MRI (1919908).</div></section><section class='html-notes'><h2 >Institutional Review Board Statement</h2><div class='html-p'>Not applicable.</div></section><section class='html-notes'><h2 >Informed Consent Statement</h2><div class='html-p'>Not applicable.</div></section><section class='html-notes'><h2 >Data Availability Statement</h2><div class='html-p'>Data are contained within the article and in the <a href="#app1-biomedicines-12-02216" class="html-app">Supplementary Materials</a>. Any additional data presented in this study are available on request from the corresponding authors.</div></section><section id='html-ack' class='html-ack'><h2 >Acknowledgments</h2><div class='html-p'>The authors recognize CSBio (20 Kelly Ct, Menlo Park, CA 94025) for the synthesis of synthetic Kalata B1.</div></section><section class='html-notes'><h2 >Conflicts of Interest</h2><div class='html-p'>The authors declare no conflicts of interest.</div></section><section id='html-references_list'><h2>References</h2><ol class='html-xx'><li id='B1-biomedicines-12-02216' class='html-x' data-content='1.'>Gran, L.; Sandberg, K.; Sletten, K. A plant containing uteroactive peptides used in African traditional medicine. <span class='html-italic'>J. Ethnopharmacol.</span> <b>2000</b>, <span class='html-italic'>70</span>, 197–203. [<a href="https://scholar.google.com/scholar_lookup?title=A+plant+containing+uteroactive+peptides+used+in+African+traditional+medicine&author=Gran,+L.&author=Sandberg,+K.&author=Sletten,+K.&publication_year=2000&journal=J.+Ethnopharmacol.&volume=70&pages=197%E2%80%93203&doi=10.1016/S0378-8741(99)00175-0" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/S0378-8741(99)00175-0" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B2-biomedicines-12-02216' class='html-x' data-content='2.'>Abdul Ghani, H.; Henriques, S.T.; Huang, Y.H.; Swedberg, J.E.; Schroeder, C.I.; Craik, D.J. Structural and functional characterization of chimeric cyclotides from the Möbius and trypsin inhibitor subfamilies. <span class='html-italic'>Biopolymers</span> <b>2017</b>, <span class='html-italic'>108</span>, e22927. [<a href="https://scholar.google.com/scholar_lookup?title=Structural+and+functional+characterization+of+chimeric+cyclotides+from+the+M%C3%B6bius+and+trypsin+inhibitor+subfamilies&author=Abdul+Ghani,+H.&author=Henriques,+S.T.&author=Huang,+Y.H.&author=Swedberg,+J.E.&author=Schroeder,+C.I.&author=Craik,+D.J.&publication_year=2017&journal=Biopolymers&volume=108&pages=e22927&doi=10.1002/bip.22927&pmid=27487329" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1002/bip.22927" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/27487329" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B3-biomedicines-12-02216' class='html-x' data-content='3.'>Jennings, C.; West, J.; Waine, C.; Craik, D.; Anderson, M. Biosynthesis and insecticidal properties of plant cyclotides: The cyclic knotted proteins from <span class='html-italic'>Oldenlandia affinis</span>. <span class='html-italic'>Proc. Natl. Acad. Sci. USA</span> <b>2001</b>, <span class='html-italic'>98</span>, 10614–10619. [<a href="https://scholar.google.com/scholar_lookup?title=Biosynthesis+and+insecticidal+properties+of+plant+cyclotides:+The+cyclic+knotted+proteins+from+Oldenlandia+affinis&author=Jennings,+C.&author=West,+J.&author=Waine,+C.&author=Craik,+D.&author=Anderson,+M.&publication_year=2001&journal=Proc.+Natl.+Acad.+Sci.+USA&volume=98&pages=10614%E2%80%9310619&doi=10.1073/pnas.191366898&pmid=11535828" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1073/pnas.191366898" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/11535828" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B4-biomedicines-12-02216' class='html-x' data-content='4.'>Gruber, C.W.; Čemažar, M.; Anderson, M.A.; Craik, D.J. Insecticidal plant cyclotides and related cystine knot toxins. <span class='html-italic'>Toxicon</span> <b>2007</b>, <span class='html-italic'>49</span>, 561–575. [<a href="https://scholar.google.com/scholar_lookup?title=Insecticidal+plant+cyclotides+and+related+cystine+knot+toxins&author=Gruber,+C.W.&author=%C4%8Cema%C5%BEar,+M.&author=Anderson,+M.A.&author=Craik,+D.J.&publication_year=2007&journal=Toxicon&volume=49&pages=561%E2%80%93575&doi=10.1016/j.toxicon.2006.11.018&pmid=17224167" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/j.toxicon.2006.11.018" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/17224167" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B5-biomedicines-12-02216' class='html-x' data-content='5.'>Strömstedt, A.A.; Park, S.; Burman, R.; Göransson, U. Bactericidal activity of cyclotides where phosphatidylethanolamine-lipid selectivity determines antimicrobial spectra. <span class='html-italic'>Biochim. Biophys. Acta Biomembr.</span> <b>2017</b>, <span class='html-italic'>1859</span>, 1986–2000. [<a href="https://scholar.google.com/scholar_lookup?title=Bactericidal+activity+of+cyclotides+where+phosphatidylethanolamine-lipid+selectivity+determines+antimicrobial+spectra&author=Str%C3%B6mstedt,+A.A.&author=Park,+S.&author=Burman,+R.&author=G%C3%B6ransson,+U.&publication_year=2017&journal=Biochim.+Biophys.+Acta+Biomembr.&volume=1859&pages=1986%E2%80%932000&doi=10.1016/j.bbamem.2017.06.018&pmid=28669767" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/j.bbamem.2017.06.018" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/28669767" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B6-biomedicines-12-02216' class='html-x' data-content='6.'>Slazak, B.; Kapusta, M.; Strömstedt, A.A.; Słomka, A.; Krychowiak, M.; Shariatgorji, M.; Andrén, P.E.; Bohdanowicz, J.; Kuta, E.; Göransson, U. How Does the Sweet Violet (<span class='html-italic'>Viola odorata</span> L.) Fight Pathogens and Pests—Cyclotides as a Comprehensive Plant Host Defense System. <span class='html-italic'>Front. Plant Sci.</span> <b>2018</b>, <span class='html-italic'>9</span>, 1296. [<a href="https://scholar.google.com/scholar_lookup?title=How+Does+the+Sweet+Violet+(Viola+odorata+L.)+Fight+Pathogens+and+Pests%E2%80%94Cyclotides+as+a+Comprehensive+Plant+Host+Defense+System&author=Slazak,+B.&author=Kapusta,+M.&author=Str%C3%B6mstedt,+A.A.&author=S%C5%82omka,+A.&author=Krychowiak,+M.&author=Shariatgorji,+M.&author=Andr%C3%A9n,+P.E.&author=Bohdanowicz,+J.&author=Kuta,+E.&author=G%C3%B6ransson,+U.&publication_year=2018&journal=Front.+Plant+Sci.&volume=9&pages=1296&doi=10.3389/fpls.2018.01296" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.3389/fpls.2018.01296" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B7-biomedicines-12-02216' class='html-x' data-content='7.'>Henriques, S.T.; Craik, D.J. Cyclotides as templates in drug design. <span class='html-italic'>Drug Discov. Today</span> <b>2010</b>, <span class='html-italic'>15</span>, 57–64. [<a href="https://scholar.google.com/scholar_lookup?title=Cyclotides+as+templates+in+drug+design&author=Henriques,+S.T.&author=Craik,+D.J.&publication_year=2010&journal=Drug+Discov.+Today&volume=15&pages=57%E2%80%9364&doi=10.1016/j.drudis.2009.10.007" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/j.drudis.2009.10.007" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B8-biomedicines-12-02216' class='html-x' data-content='8.'>Azmi, S.; Mustafa, M.; Shoaib, S.; Hussain, M.K. Structures, Functions and Therapeutic Potential of Cyclotides. <span class='html-italic'>J. Explor. Res. Pharm.</span> <b>2022</b>, <span class='html-italic'>7</span>, 234–242. [<a href="https://scholar.google.com/scholar_lookup?title=Structures,+Functions+and+Therapeutic+Potential+of+Cyclotides&author=Azmi,+S.&author=Mustafa,+M.&author=Shoaib,+S.&author=Hussain,+M.K.&publication_year=2022&journal=J.+Explor.+Res.+Pharm.&volume=7&pages=234%E2%80%93242&doi=10.14218/JERP.2022.00029" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.14218/JERP.2022.00029" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B9-biomedicines-12-02216' class='html-x' data-content='9.'>Gerlach, S.L.; Dunlop, R.A.; Metcalf, J.S.; Banack, S.A.; Cox, P.A. Cyclotides Chemosensitize Glioblastoma Cells to Temozolomide. <span class='html-italic'>J. Nat. Prod.</span> <b>2022</b>, <span class='html-italic'>85</span>, 34–46. [<a href="https://scholar.google.com/scholar_lookup?title=Cyclotides+Chemosensitize+Glioblastoma+Cells+to+Temozolomide&author=Gerlach,+S.L.&author=Dunlop,+R.A.&author=Metcalf,+J.S.&author=Banack,+S.A.&author=Cox,+P.A.&publication_year=2022&journal=J.+Nat.+Prod.&volume=85&pages=34%E2%80%9346&doi=10.1021/acs.jnatprod.1c00595" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1021/acs.jnatprod.1c00595" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B10-biomedicines-12-02216' class='html-xx' data-content='10.'>Ireland, D.C.; Wang, C.K.; Wilson, J.A.; Gustafson, K.R.; Craik, D.J. Cyclotides as natural anti-HIV agents. <span class='html-italic'>Biopolymers</span> <b>2008</b>, <span class='html-italic'>90</span>, 51–60. [<a href="https://scholar.google.com/scholar_lookup?title=Cyclotides+as+natural+anti-HIV+agents&author=Ireland,+D.C.&author=Wang,+C.K.&author=Wilson,+J.A.&author=Gustafson,+K.R.&author=Craik,+D.J.&publication_year=2008&journal=Biopolymers&volume=90&pages=51%E2%80%9360&doi=10.1002/bip.20886" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1002/bip.20886" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B11-biomedicines-12-02216' class='html-xx' data-content='11.'>Gerlach, S.L.; Rathinakumar, R.; Chakravarty, G.; Göransson, U.; Wimley, W.C.; Darwin, S.P.; Mondal, D. Anticancer and chemosensitizing abilities of cycloviolacin 02 from Viola odorata and psyle cyclotides from <span class='html-italic'>Psychotria leptothyrsa</span>. <span class='html-italic'>Biopolymers</span> <b>2010</b>, <span class='html-italic'>94</span>, 617–625. [<a href="https://scholar.google.com/scholar_lookup?title=Anticancer+and+chemosensitizing+abilities+of+cycloviolacin+02+from+Viola+odorata+and+psyle+cyclotides+from+Psychotria+leptothyrsa&author=Gerlach,+S.L.&author=Rathinakumar,+R.&author=Chakravarty,+G.&author=G%C3%B6ransson,+U.&author=Wimley,+W.C.&author=Darwin,+S.P.&author=Mondal,+D.&publication_year=2010&journal=Biopolymers&volume=94&pages=617%E2%80%93625&doi=10.1002/bip.21435&pmid=20564026" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1002/bip.21435" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/20564026" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B12-biomedicines-12-02216' class='html-xx' data-content='12.'>Gerlach, S.L.; Chandra, P.K.; Gunasekera, S.; Göransson u Wimley, W.C.; Braun, S.E.; Mondal, D. The membrane-active phytopeptide cycloviolacin O<sub>2</sub> targets HIV-1 infected cells and infectious viral particles to potentiate the efficacy of antiretroviral drugs. <span class='html-italic'>Medicines</span> <b>2019</b>, <span class='html-italic'>6</span>, 33. [<a href="https://scholar.google.com/scholar_lookup?title=The+membrane-active+phytopeptide+cycloviolacin+O2+targets+HIV-1+infected+cells+and+infectious+viral+particles+to+potentiate+the+efficacy+of+antiretroviral+drugs&author=Gerlach,+S.L.&author=Chandra,+P.K.&author=Gunasekera,+S.&author=G%C3%B6ransson+u+Wimley,+W.C.&author=Braun,+S.E.&author=Mondal,+D.&publication_year=2019&journal=Medicines&volume=6&pages=33&doi=10.3390/medicines6010033&pmid=30823453" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.3390/medicines6010033" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/30823453" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B13-biomedicines-12-02216' class='html-xx' data-content='13.'>Janjua, T.I.; Rewatkar, P.; Ahmed-Cox, A.; Saeed, I.; Mansfeld, F.M.; Kulshreshtha, R.; Kumeria, T.; Ziegler, D.S.; Kavallaris, M.; Mazzieri, R.; et al. Frontiers in the treatment of glioblastoma: Past, present, and emerging. <span class='html-italic'>Adv. Drug Deliv. Rev.</span> <b>2021</b>, <span class='html-italic'>171</span>, 108–138. [<a href="https://scholar.google.com/scholar_lookup?title=Frontiers+in+the+treatment+of+glioblastoma:+Past,+present,+and+emerging&author=Janjua,+T.I.&author=Rewatkar,+P.&author=Ahmed-Cox,+A.&author=Saeed,+I.&author=Mansfeld,+F.M.&author=Kulshreshtha,+R.&author=Kumeria,+T.&author=Ziegler,+D.S.&author=Kavallaris,+M.&author=Mazzieri,+R.&publication_year=2021&journal=Adv.+Drug+Deliv.+Rev.&volume=171&pages=108%E2%80%93138&doi=10.1016/j.addr.2021.01.012&pmid=33486006" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/j.addr.2021.01.012" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/33486006" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B14-biomedicines-12-02216' class='html-xx' data-content='14.'>Wu, W.; Klockow, J.L.; Zhang, M.; Lafortune, F.; Chang, E.; Jin, L.; Wu, Y.; Daldrup-Link, H.E. Glioblastoma multiforme (GBM): An overview of current therapies and mechanisms of resistance. <span class='html-italic'>Pharmacol. Res.</span> <b>2021</b>, <span class='html-italic'>171</span>, 105780. [<a href="https://scholar.google.com/scholar_lookup?title=Glioblastoma+multiforme+(GBM):+An+overview+of+current+therapies+and+mechanisms+of+resistance&author=Wu,+W.&author=Klockow,+J.L.&author=Zhang,+M.&author=Lafortune,+F.&author=Chang,+E.&author=Jin,+L.&author=Wu,+Y.&author=Daldrup-Link,+H.E.&publication_year=2021&journal=Pharmacol.+Res.&volume=171&pages=105780&doi=10.1016/j.phrs.2021.105780&pmid=34302977" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/j.phrs.2021.105780" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/34302977" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B15-biomedicines-12-02216' class='html-xx' data-content='15.'>Stupp, R.; Mason, W.P.; van den Bent, M.J.; Weller, M.; Fisher, B.; Taphoorn, M.J.B.; Belanger, K.; Brandes, A.A.; Marosi, C.; Bogdahn, U.; et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. <span class='html-italic'>N. Engl. J. Med.</span> <b>2005</b>, <span class='html-italic'>352</span>, 987–996. [<a href="https://scholar.google.com/scholar_lookup?title=Radiotherapy+plus+concomitant+and+adjuvant+temozolomide+for+glioblastoma&author=Stupp,+R.&author=Mason,+W.P.&author=van+den+Bent,+M.J.&author=Weller,+M.&author=Fisher,+B.&author=Taphoorn,+M.J.B.&author=Belanger,+K.&author=Brandes,+A.A.&author=Marosi,+C.&author=Bogdahn,+U.&publication_year=2005&journal=N.+Engl.+J.+Med.&volume=352&pages=987%E2%80%93996&doi=10.1056/NEJMoa043330" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1056/NEJMoa043330" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B16-biomedicines-12-02216' class='html-xx' data-content='16.'>Singh, N.; Miner, A.; Hennis, L.; Mittal, S. Mechanisms of temozolomide resistance in glioblastoma—A comprehensive review. <span class='html-italic'>Cancer Drug Resist.</span> <b>2021</b>, <span class='html-italic'>4</span>, 17–43. [<a href="https://scholar.google.com/scholar_lookup?title=Mechanisms+of+temozolomide+resistance+in+glioblastoma%E2%80%94A+comprehensive+review&author=Singh,+N.&author=Miner,+A.&author=Hennis,+L.&author=Mittal,+S.&publication_year=2021&journal=Cancer+Drug+Resist.&volume=4&pages=17%E2%80%9343&doi=10.20517/cdr.2020.79" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.20517/cdr.2020.79" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B17-biomedicines-12-02216' class='html-xx' data-content='17.'>Rajendran, S.; Slazak, B.; Mohotti, S.; Muhammad, T.; Strömstedt, A.A.; Kapusta, M.M.; Wilmowicz, E.; Göransson, U.; Hettiarachchi, C.M.; Gunasekera, S. Screening for Cyclotides in Sri Lankan Medicinal Plants: Discovery, Characterization, and Bioactivity Screening of Cyclotides from <span class='html-italic'>Geophila repens</span>. <span class='html-italic'>J. Nat. Prod.</span> <b>2023</b>, <span class='html-italic'>86</span>, 52–65. [<a href="https://scholar.google.com/scholar_lookup?title=Screening+for+Cyclotides+in+Sri+Lankan+Medicinal+Plants:+Discovery,+Characterization,+and+Bioactivity+Screening+of+Cyclotides+from+Geophila+repens&author=Rajendran,+S.&author=Slazak,+B.&author=Mohotti,+S.&author=Muhammad,+T.&author=Str%C3%B6mstedt,+A.A.&author=Kapusta,+M.M.&author=Wilmowicz,+E.&author=G%C3%B6ransson,+U.&author=Hettiarachchi,+C.M.&author=Gunasekera,+S.&publication_year=2023&journal=J.+Nat.+Prod.&volume=86&pages=52%E2%80%9365&doi=10.1021/acs.jnatprod.2c00674" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1021/acs.jnatprod.2c00674" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B18-biomedicines-12-02216' class='html-xx' data-content='18.'>Rance, M.O.; Sørensen, G.; Bodenhausen, G.; Wagner, R.R.; Wüthrich, K. Improved spectral resolution in COSY 1H NMR spectra of proteins via double quantum filtering. <span class='html-italic'>Biochem. Biophys. Res. Commun.</span> <b>1983</b>, <span class='html-italic'>117</span>, 479–485. [<a href="https://scholar.google.com/scholar_lookup?title=Improved+spectral+resolution+in+COSY+1H+NMR+spectra+of+proteins+via+double+quantum+filtering&author=Rance,+M.O.&author=S%C3%B8rensen,+G.&author=Bodenhausen,+G.&author=Wagner,+R.R.&author=W%C3%BCthrich,+K.&publication_year=1983&journal=Biochem.+Biophys.+Res.+Commun.&volume=117&pages=479%E2%80%93485&doi=10.1016/0006-291X(83)91225-1" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/0006-291X(83)91225-1" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B19-biomedicines-12-02216' class='html-xx' data-content='19.'>Rucker, S.P.; Shaka, A. Broadband homonuclear cross polarization in 2D NMR using DIPSI-2. <span class='html-italic'>Mol. Phys.</span> <b>1989</b>, <span class='html-italic'>68</span>, 509–517. [<a href="https://scholar.google.com/scholar_lookup?title=Broadband+homonuclear+cross+polarization+in+2D+NMR+using+DIPSI-2&author=Rucker,+S.P.&author=Shaka,+A.&publication_year=1989&journal=Mol.+Phys.&volume=68&pages=509%E2%80%93517&doi=10.1080/00268978900102331" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1080/00268978900102331" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B20-biomedicines-12-02216' class='html-xx' data-content='20.'>Bothner-By, A.A.; Stephens, R.; Lee, J.; Warren, C.D.; Jeanloz, R. Structure determination of a tetrasaccharide: Transient nuclear Overhauser effects in the rotating frame. <span class='html-italic'>J. Am. Chem. Soc.</span> <b>1984</b>, <span class='html-italic'>106</span>, 811–813. [<a href="https://scholar.google.com/scholar_lookup?title=Structure+determination+of+a+tetrasaccharide:+Transient+nuclear+Overhauser+effects+in+the+rotating+frame&author=Bothner-By,+A.A.&author=Stephens,+R.&author=Lee,+J.&author=Warren,+C.D.&author=Jeanloz,+R.&publication_year=1984&journal=J.+Am.+Chem.+Soc.&volume=106&pages=811%E2%80%93813&doi=10.1021/ja00315a069" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1021/ja00315a069" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B21-biomedicines-12-02216' class='html-xx' data-content='21.'>Bax, A.; Davis, D.G. Practical aspects of two-dimensional transverse NOE spectroscopy. <span class='html-italic'>J. Mag. Reson.</span> <b>1985</b>, <span class='html-italic'>63</span>, 207–213. [<a href="https://scholar.google.com/scholar_lookup?title=Practical+aspects+of+two-dimensional+transverse+NOE+spectroscopy&author=Bax,+A.&author=Davis,+D.G.&publication_year=1985&journal=J.+Mag.+Reson.&volume=63&pages=207%E2%80%93213&doi=10.1016/0022-2364(85)90171-4" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/0022-2364(85)90171-4" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B22-biomedicines-12-02216' class='html-xx' data-content='22.'>Kumar, A.; Ernes, R.; Wüthrich, K. A two-dimensional nuclear Overhauser enhancement (2D NOE) experiment for the elucidation of complete proton-proton cross-relaxation networks in biological macromolecules. <span class='html-italic'>Biochem. Biophys. Res. Commun.</span> <b>1980</b>, <span class='html-italic'>95</span>, 1–6. [<a href="https://scholar.google.com/scholar_lookup?title=A+two-dimensional+nuclear+Overhauser+enhancement+(2D+NOE)+experiment+for+the+elucidation+of+complete+proton-proton+cross-relaxation+networks+in+biological+macromolecules&author=Kumar,+A.&author=Ernes,+R.&author=W%C3%BCthrich,+K.&publication_year=1980&journal=Biochem.+Biophys.+Res.+Commun.&volume=95&pages=1%E2%80%936&doi=10.1016/0006-291X(80)90695-6&pmid=7417242" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/0006-291X(80)90695-6" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/7417242" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B23-biomedicines-12-02216' class='html-xx' data-content='23.'>Goddard, T.D.; Kneller, D.G. <span class='html-italic'>SPARKY 3</span>; University of California: San Francisco, CA, USA, 2000. [<a href="https://scholar.google.com/scholar_lookup?title=SPARKY+3&author=Goddard,+T.D.&author=Kneller,+D.G.&publication_year=2000" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>]</li><li id='B24-biomedicines-12-02216' class='html-xx' data-content='24.'>Lee, W.; Tonelli, M.; Markley, J.L. NMRFAM-SPARKY: Enhancement software for biomolecular NMR spectroscopy. <span class='html-italic'>Bioinformatics</span> <b>2015</b>, <span class='html-italic'>31</span>, 1325–1327. [<a href="https://scholar.google.com/scholar_lookup?title=NMRFAM-SPARKY:+Enhancement+software+for+biomolecular+NMR+spectroscopy&author=Lee,+W.&author=Tonelli,+M.&author=Markley,+J.L.&publication_year=2015&journal=Bioinformatics&volume=31&pages=1325%E2%80%931327&doi=10.1093/bioinformatics/btu830&pmid=25505092" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1093/bioinformatics/btu830" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/25505092" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B25-biomedicines-12-02216' class='html-xx' data-content='25.'>Guntert, P.A. Automated NMR structure calculation with CYANA. <span class='html-italic'>Methods Mol. Biol.</span> <b>2004</b>, <span class='html-italic'>278</span>, 353–378. [<a href="https://scholar.google.com/scholar_lookup?title=Automated+NMR+structure+calculation+with+CYANA&author=Guntert,+P.A.&publication_year=2004&journal=Methods+Mol.+Biol.&volume=278&pages=353%E2%80%93378&pmid=15318003" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/15318003" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B26-biomedicines-12-02216' class='html-xx' data-content='26.'>Mosmann, T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. <span class='html-italic'>J. Immunol. Methods</span> <b>1983</b>, <span class='html-italic'>65</span>, 55–63. [<a href="https://scholar.google.com/scholar_lookup?title=Rapid+colorimetric+assay+for+cellular+growth+and+survival:+Application+to+proliferation+and+cytotoxicity+assays&author=Mosmann,+T.&publication_year=1983&journal=J.+Immunol.+Methods&volume=65&pages=55%E2%80%9363&doi=10.1016/0022-1759(83)90303-4" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/0022-1759(83)90303-4" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B27-biomedicines-12-02216' class='html-xx' data-content='27.'>Chan, L.Y.; Gunasekera, S.; Henriques, S.T.; Worth, N.F.; Le, S.-J.; Clark, R.J.; Campbell, J.H.; Craik, D.J.; Daly, M.L. Engineering pro-angiogenic peptides using stable, disulfide-rich cyclic scaffolds. <span class='html-italic'>Blood.</span> <b>2011</b>, <span class='html-italic'>118</span>, 6709–6717. [<a href="https://scholar.google.com/scholar_lookup?title=Engineering+pro-angiogenic+peptides+using+stable,+disulfide-rich+cyclic+scaffolds&author=Chan,+L.Y.&author=Gunasekera,+S.&author=Henriques,+S.T.&author=Worth,+N.F.&author=Le,+S.-J.&author=Clark,+R.J.&author=Campbell,+J.H.&author=Craik,+D.J.&author=Daly,+M.L.&publication_year=2011&journal=Blood.&volume=118&pages=6709%E2%80%936717&doi=10.1182/blood-2011-06-359141" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1182/blood-2011-06-359141" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B28-biomedicines-12-02216' class='html-xx' data-content='28.'>Huang, Y.H.; Chaousis, S.; Cheneval, O.; Craik, D.J.; Henriques, S.T. Optimization of the cyclotide framework to improve cell penetration properties. <span class='html-italic'>Front. Pharmacol.</span> <b>2015</b>, <span class='html-italic'>6</span>, 17. [<a href="https://scholar.google.com/scholar_lookup?title=Optimization+of+the+cyclotide+framework+to+improve+cell+penetration+properties&author=Huang,+Y.H.&author=Chaousis,+S.&author=Cheneval,+O.&author=Craik,+D.J.&author=Henriques,+S.T.&publication_year=2015&journal=Front.+Pharmacol.&volume=6&pages=17&doi=10.3389/fphar.2015.00017" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.3389/fphar.2015.00017" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B29-biomedicines-12-02216' class='html-xx' data-content='29.'>Wüthrich, K. <span class='html-italic'>NMR of Proteins and Nucleic Acids</span>; John Wiley & Sons, Inc.: Hoboken, NJ, USA, 1986. [<a href="https://scholar.google.com/scholar_lookup?title=NMR+of+Proteins+and+Nucleic+Acids&author=W%C3%BCthrich,+K.&publication_year=1986" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>]</li><li id='B30-biomedicines-12-02216' class='html-xx' data-content='30.'>Niyomploy, P.; Chan, L.Y.; Poth, A.G.; Colgrave, M.L.; Sangvanich, P.; Craik, D.J. Discovery, isolation, and structural characterization of cyclotides from <span class='html-italic'>Viola sumatrana</span> Miq. <span class='html-italic'>Biopolymers</span> <b>2016</b>, <span class='html-italic'>106</span>, 796–805. [<a href="https://scholar.google.com/scholar_lookup?title=Discovery,+isolation,+and+structural+characterization+of+cyclotides+from+Viola+sumatrana+Miq&author=Niyomploy,+P.&author=Chan,+L.Y.&author=Poth,+A.G.&author=Colgrave,+M.L.&author=Sangvanich,+P.&author=Craik,+D.J.&publication_year=2016&journal=Biopolymers&volume=106&pages=796%E2%80%93805&doi=10.1002/bip.22914" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1002/bip.22914" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B31-biomedicines-12-02216' class='html-xx' data-content='31.'>Young, R.M.; Jamshidi, A.; Davis, G.; Sherman, J.H. Current trends in the surgical management and treatment of adult glioblastoma. <span class='html-italic'>Ann. Transl. Med.</span> <b>2015</b>, <span class='html-italic'>3</span>, 121. [<a href="https://scholar.google.com/scholar_lookup?title=Current+trends+in+the+surgical+management+and+treatment+of+adult+glioblastoma&author=Young,+R.M.&author=Jamshidi,+A.&author=Davis,+G.&author=Sherman,+J.H.&publication_year=2015&journal=Ann.+Transl.+Med.&volume=3&pages=121&doi=10.3978/j.issn.2305-5839.2015.05.10" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.3978/j.issn.2305-5839.2015.05.10" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B32-biomedicines-12-02216' class='html-xx' data-content='32.'>Lee, S.Y. Temozolomide resistance in glioblastoma multiforme. <span class='html-italic'>Genes Dis.</span> <b>2016</b>, <span class='html-italic'>3</span>, 198–210. [<a href="https://scholar.google.com/scholar_lookup?title=Temozolomide+resistance+in+glioblastoma+multiforme&author=Lee,+S.Y.&publication_year=2016&journal=Genes+Dis.&volume=3&pages=198%E2%80%93210&doi=10.1016/j.gendis.2016.04.007" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/j.gendis.2016.04.007" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B33-biomedicines-12-02216' class='html-xx' data-content='33.'>Bi, W.L.; Beroukhim, R. Beating the odds: Extreme long-term survival with glioblastoma. <span class='html-italic'>Neuro-Oncology</span> <b>2014</b>, <span class='html-italic'>16</span>, 1159–1160. [<a href="https://scholar.google.com/scholar_lookup?title=Beating+the+odds:+Extreme+long-term+survival+with+glioblastoma&author=Bi,+W.L.&author=Beroukhim,+R.&publication_year=2014&journal=Neuro-Oncology&volume=16&pages=1159%E2%80%931160&doi=10.1093/neuonc/nou166&pmid=25096192" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1093/neuonc/nou166" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/25096192" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B34-biomedicines-12-02216' class='html-xx' data-content='34.'>Tang, J.; Wang, C.K.; Pan, X.; Yan, H.; Zeng, G.; Xu, W.; He, W.; Daly, N.L.; Craik, D.J.; Tan, N. Isolation and characterization of cytotoxic cyclotides from Viola tricolor. <span class='html-italic'>Peptides</span> <b>2010</b>, <span class='html-italic'>31</span>, 1434–1440. [<a href="https://scholar.google.com/scholar_lookup?title=Isolation+and+characterization+of+cytotoxic+cyclotides+from+Viola+tricolor&author=Tang,+J.&author=Wang,+C.K.&author=Pan,+X.&author=Yan,+H.&author=Zeng,+G.&author=Xu,+W.&author=He,+W.&author=Daly,+N.L.&author=Craik,+D.J.&author=Tan,+N.&publication_year=2010&journal=Peptides&volume=31&pages=1434%E2%80%931440&doi=10.1016/j.peptides.2010.05.004&pmid=20580652" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/j.peptides.2010.05.004" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/20580652" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B35-biomedicines-12-02216' class='html-xx' data-content='35.'>Henriques, S.T.; Huang, Y.H.; Chaousis, S.; Sani, M.-A.; Poth, A.G.; Separovic, F.; Craik, D.J. The Prototypic Cyclotide Kalata B1 Has a Unique Mechanism of Entering Cells. <span class='html-italic'>Chem. Biol.</span> <b>2015</b>, <span class='html-italic'>22</span>, 1087–1097. [<a href="https://scholar.google.com/scholar_lookup?title=The+Prototypic+Cyclotide+Kalata+B1+Has+a+Unique+Mechanism+of+Entering+Cells&author=Henriques,+S.T.&author=Huang,+Y.H.&author=Chaousis,+S.&author=Sani,+M.-A.&author=Poth,+A.G.&author=Separovic,+F.&author=Craik,+D.J.&publication_year=2015&journal=Chem.+Biol.&volume=22&pages=1087%E2%80%931097&doi=10.1016/j.chembiol.2015.07.012&pmid=26278183" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/j.chembiol.2015.07.012" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/26278183" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B36-biomedicines-12-02216' class='html-xx' data-content='36.'>Lindholm, P.; Göransson, U.; Johansson, S.; Claeson, P.; Gullbo, J.; Larsson, R.; Bohlin, L.; Backlund, A. Cyclotides: A novel type of cytotoxic agents. <span class='html-italic'>Mol. Cancer Ther.</span> <b>2002</b>, <span class='html-italic'>1</span>, 365–369. [<a href="https://scholar.google.com/scholar_lookup?title=Cyclotides:+A+novel+type+of+cytotoxic+agents&author=Lindholm,+P.&author=G%C3%B6ransson,+U.&author=Johansson,+S.&author=Claeson,+P.&author=Gullbo,+J.&author=Larsson,+R.&author=Bohlin,+L.&author=Backlund,+A.&publication_year=2002&journal=Mol.+Cancer+Ther.&volume=1&pages=365%E2%80%93369&pmid=12477048" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/12477048" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B37-biomedicines-12-02216' class='html-xx' data-content='37.'>Hall, K.; Lee, T.H.; Daly, N.L.; Craik, D.J.; Aguilar, M.I. Gly (6) of kalata B1 is critical for the selective binding to phosphatidylethanolamine membranes. <span class='html-italic'>Biochim. Biophys. Acta</span> <b>2012</b>, <span class='html-italic'>1818</span>, 2354–2361. [<a href="https://scholar.google.com/scholar_lookup?title=Gly+(6)+of+kalata+B1+is+critical+for+the+selective+binding+to+phosphatidylethanolamine+membranes&author=Hall,+K.&author=Lee,+T.H.&author=Daly,+N.L.&author=Craik,+D.J.&author=Aguilar,+M.I.&publication_year=2012&journal=Biochim.+Biophys.+Acta&volume=1818&pages=2354%E2%80%932361&doi=10.1016/j.bbamem.2012.04.007" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/j.bbamem.2012.04.007" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B38-biomedicines-12-02216' class='html-xx' data-content='38.'>Cransfield, C.G.; Henriques, S.T.; Martinac, B.; Duckworth, P.; Craik, D.J.; Cornell, B. Kalata B1 and Kalata B2 have a surfactant-like activity in phosphatidylethanolamine-containing lipid membranes. <span class='html-italic'>Langmuir</span> <b>2017</b>, <span class='html-italic'>33</span>, 6630–6637. [<a href="https://scholar.google.com/scholar_lookup?title=Kalata+B1+and+Kalata+B2+have+a+surfactant-like+activity+in+phosphatidylethanolamine-containing+lipid+membranes&author=Cransfield,+C.G.&author=Henriques,+S.T.&author=Martinac,+B.&author=Duckworth,+P.&author=Craik,+D.J.&author=Cornell,+B.&publication_year=2017&journal=Langmuir&volume=33&pages=6630%E2%80%936637&doi=10.1021/acs.langmuir.7b01642" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1021/acs.langmuir.7b01642" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B39-biomedicines-12-02216' class='html-xx' data-content='39.'>Gründemann, C.; Thell, K.; Lengen, K.; Garcia-Käufer, M.; Huang, Y.-H.; Huber, R.; Craik, D.J.; Schabbauer, G.; Gruber, C.W. Cyclotides suppress human T-lymphocyte proliferation by an interleukin 2-dependent mechanism. <span class='html-italic'>PLoS ONE</span> <b>2013</b>, <span class='html-italic'>8</span>, 68016. [<a href="https://scholar.google.com/scholar_lookup?title=Cyclotides+suppress+human+T-lymphocyte+proliferation+by+an+interleukin+2-dependent+mechanism&author=Gr%C3%BCndemann,+C.&author=Thell,+K.&author=Lengen,+K.&author=Garcia-K%C3%A4ufer,+M.&author=Huang,+Y.-H.&author=Huber,+R.&author=Craik,+D.J.&author=Schabbauer,+G.&author=Gruber,+C.W.&publication_year=2013&journal=PLoS+ONE&volume=8&pages=68016&doi=10.1371/journal.pone.0068016" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1371/journal.pone.0068016" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B40-biomedicines-12-02216' class='html-xx' data-content='40.'>Burman, S.; Svedlund, F.; Felth, J.; Hassan, S.; Herrmann, A.; Clark, R.J.; Craik, D.J.; Bohlin, L.; Claeson, P.; Göransson, U.; et al. Evaluation of toxicity and antitumor activity of cycloviolacin O<sub>2</sub> in mice. <span class='html-italic'>Biopolymers</span> <b>2010</b>, <span class='html-italic'>94</span>, 626–634. [<a href="https://scholar.google.com/scholar_lookup?title=Evaluation+of+toxicity+and+antitumor+activity+of+cycloviolacin+O2+in+mice&author=Burman,+S.&author=Svedlund,+F.&author=Felth,+J.&author=Hassan,+S.&author=Herrmann,+A.&author=Clark,+R.J.&author=Craik,+D.J.&author=Bohlin,+L.&author=Claeson,+P.&author=G%C3%B6ransson,+U.&publication_year=2010&journal=Biopolymers&volume=94&pages=626%E2%80%93634&doi=10.1002/bip.21408" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1002/bip.21408" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B41-biomedicines-12-02216' class='html-xx' data-content='41.'>Lind, J.; Hellinger, R.; Kudweis, P.; Moll, H.P.; Gattringer, J.; Thell, K.; Edtmayer, S.; Gruber, C.W.; Stoiber, D.; Kollmann, K. The nature inspired peptide [T20k]-kalata B1 induces anti-tumor effects in anaplastic large cell lymphoma. <span class='html-italic'>Biomed. Pharmacother.</span> <b>2022</b>, <span class='html-italic'>153</span>, 113486. [<a href="https://scholar.google.com/scholar_lookup?title=The+nature+inspired+peptide+%5BT20k%5D-kalata+B1+induces+anti-tumor+effects+in+anaplastic+large+cell+lymphoma&author=Lind,+J.&author=Hellinger,+R.&author=Kudweis,+P.&author=Moll,+H.P.&author=Gattringer,+J.&author=Thell,+K.&author=Edtmayer,+S.&author=Gruber,+C.W.&author=Stoiber,+D.&author=Kollmann,+K.&publication_year=2022&journal=Biomed.+Pharmacother.&volume=153&pages=113486&doi=10.1016/j.biopha.2022.113486" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1016/j.biopha.2022.113486" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B42-biomedicines-12-02216' class='html-xx' data-content='42.'>Wang, C.K.; Stalmans, S.; De Spiegeleer, B.; Craik, D.J. Biodistribution of the cyclotide MCoTI-II, a cyclic disulfide-rich peptide drug scaffold. <span class='html-italic'>J. Pept. Sci.</span> <b>2016</b>, <span class='html-italic'>22</span>, 305–310. [<a href="https://scholar.google.com/scholar_lookup?title=Biodistribution+of+the+cyclotide+MCoTI-II,+a+cyclic+disulfide-rich+peptide+drug+scaffold&author=Wang,+C.K.&author=Stalmans,+S.&author=De+Spiegeleer,+B.&author=Craik,+D.J.&publication_year=2016&journal=J.+Pept.+Sci.&volume=22&pages=305%E2%80%93310&doi=10.1002/psc.2862" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1002/psc.2862" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>]</li><li id='B43-biomedicines-12-02216' class='html-xx' data-content='43.'>Hernandez, J.; Gagnon, J.; Chiche, L.; Nguyen, T.; Andrieu, J.; Heitz, A.; Hong, T.T.; Pham, T.; Nguyen, D.L. Squash trypsin inhibitors from Momordica cochinchinensis exhibit an atypical macrocyclic structure. <span class='html-italic'>Biochemistry</span> <b>2002</b>, <span class='html-italic'>39</span>, 5722–5730. [<a href="https://scholar.google.com/scholar_lookup?title=Squash+trypsin+inhibitors+from+Momordica+cochinchinensis+exhibit+an+atypical+macrocyclic+structure&author=Hernandez,+J.&author=Gagnon,+J.&author=Chiche,+L.&author=Nguyen,+T.&author=Andrieu,+J.&author=Heitz,+A.&author=Hong,+T.T.&author=Pham,+T.&author=Nguyen,+D.L.&publication_year=2002&journal=Biochemistry&volume=39&pages=5722%E2%80%935730&doi=10.1021/bi9929756&pmid=10801322" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1021/bi9929756" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/10801322" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B44-biomedicines-12-02216' class='html-xx' data-content='44.'>Lesniak, W.G.; Aboye, T.; Chatterjee, S.; Camarero, J.A.; Nimmagadda, S. In vivo Evaluation of an Engineered Cyclotide as Specific CXCR4 Imaging Reagent. <span class='html-italic'>Chemistry</span> <b>2017</b>, <span class='html-italic'>23</span>, 14469–14475. [<a href="https://scholar.google.com/scholar_lookup?title=In+vivo+Evaluation+of+an+Engineered+Cyclotide+as+Specific+CXCR4+Imaging+Reagent&author=Lesniak,+W.G.&author=Aboye,+T.&author=Chatterjee,+S.&author=Camarero,+J.A.&author=Nimmagadda,+S.&publication_year=2017&journal=Chemistry&volume=23&pages=14469%E2%80%9314475&doi=10.1002/chem.201702540&pmid=28771849" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1002/chem.201702540" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/28771849" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li><li id='B45-biomedicines-12-02216' class='html-xx' data-content='45.'>Felizmenio-Quimio, M.E.; Daly, N.L.; Craik, D.J. Circular proteins in plants: Solution structure of a novel macrocyclic trypsin inhibitor from Mormordica cochinchinensis. <span class='html-italic'>J. Biol. Chem.</span> <b>2001</b>, <span class='html-italic'>276</span>, 22875–22882. [<a href="https://scholar.google.com/scholar_lookup?title=Circular+proteins+in+plants:+Solution+structure+of+a+novel+macrocyclic+trypsin+inhibitor+from+Mormordica+cochinchinensis&author=Felizmenio-Quimio,+M.E.&author=Daly,+N.L.&author=Craik,+D.J.&publication_year=2001&journal=J.+Biol.+Chem.&volume=276&pages=22875%E2%80%9322882&doi=10.1074/jbc.M101666200&pmid=11292835" class='google-scholar' target='_blank' rel='noopener noreferrer'>Google Scholar</a>] [<a href="https://doi.org/10.1074/jbc.M101666200" class='cross-ref' target='_blank' rel='noopener noreferrer'>CrossRef</a>] [<a href="https://www.ncbi.nlm.nih.gov/pubmed/11292835" class='cross-ref pub_med' data-typ='pmid' target='_blank' rel='noopener noreferrer'>PubMed</a>]</li></ol></section><section id='FiguresandTables' type='display-objects'><div class="html-fig-wrap" id="biomedicines-12-02216-f001"> <div class='html-fig_img'> <div class="html-figpopup html-figpopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f001"> <img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g001.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g001.png" alt="Biomedicines 12 02216 g001" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g001-550.jpg" /> <a class="html-expand html-figpopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f001"></a> </div> </div> <div class="html-fig_description"> <b>Figure 1.</b> A representation of the structure and sequences of cyclotides. Panel (<b>A</b>) is a three-dimensional ribbon image of Kalata B1 (PDB 1NB1, SwissProt P56254), while panel (<b>B</b>) is a circular representation of Kalata B1. Panel (<b>C</b>) depicts a ball and stick model created in JSmol. The conserved cysteine residues are highlighted in yellow, while the disulfide bonds are depicted by the red bars in panel (<b>B</b>). Number of amino acids (AA), average molecular weights (ave. mass), and the sequence of amino acids are provided for cyclotides evaluated (panel (<b>D</b>)).  </div> </div> <div class="html-fig_show mfp-hide" id="fig_body_display_biomedicines-12-02216-f001"> <div class="html-caption"> <b>Figure 1.</b> A representation of the structure and sequences of cyclotides. Panel (<b>A</b>) is a three-dimensional ribbon image of Kalata B1 (PDB 1NB1, SwissProt P56254), while panel (<b>B</b>) is a circular representation of Kalata B1. Panel (<b>C</b>) depicts a ball and stick model created in JSmol. The conserved cysteine residues are highlighted in yellow, while the disulfide bonds are depicted by the red bars in panel (<b>B</b>). Number of amino acids (AA), average molecular weights (ave. mass), and the sequence of amino acids are provided for cyclotides evaluated (panel (<b>D</b>)).</div> <div class="html-img"><img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g001.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g001.png" alt="Biomedicines 12 02216 g001" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g001.png" /></div> </div> <div class="html-fig-wrap" id="biomedicines-12-02216-f002"> <div class='html-fig_img'> <div class="html-figpopup html-figpopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f002"> <img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g002.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g002.png" alt="Biomedicines 12 02216 g002" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g002-550.jpg" /> <a class="html-expand html-figpopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f002"></a> </div> </div> <div class="html-fig_description"> <b>Figure 2.</b> UPLC-MS of synthetic Kalata B1 (black peak) and natural Kalata B1 (red peak), each with a <span class='html-italic'>m</span>/<span class='html-italic'>z</span> of 964.77 and an elution time of 5.05–5.25 min.  </div> </div> <div class="html-fig_show mfp-hide" id="fig_body_display_biomedicines-12-02216-f002"> <div class="html-caption"> <b>Figure 2.</b> UPLC-MS of synthetic Kalata B1 (black peak) and natural Kalata B1 (red peak), each with a <span class='html-italic'>m</span>/<span class='html-italic'>z</span> of 964.77 and an elution time of 5.05–5.25 min.</div> <div class="html-img"><img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g002.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g002.png" alt="Biomedicines 12 02216 g002" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g002.png" /></div> </div> <div class="html-fig-wrap" id="biomedicines-12-02216-f003"> <div class='html-fig_img'> <div class="html-figpopup html-figpopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f003"> <img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g003.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g003.png" alt="Biomedicines 12 02216 g003" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g003-550.jpg" /> <a class="html-expand html-figpopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f003"></a> </div> </div> <div class="html-fig_description"> <b>Figure 3.</b> MS/MS analysis of peptide 2 (RTCVP) with a mass of <span class='html-italic'>m</span>/<span class='html-italic'>z</span> 1215.9891.  </div> </div> <div class="html-fig_show mfp-hide" id="fig_body_display_biomedicines-12-02216-f003"> <div class="html-caption"> <b>Figure 3.</b> MS/MS analysis of peptide 2 (RTCVP) with a mass of <span class='html-italic'>m</span>/<span class='html-italic'>z</span> 1215.9891.</div> <div class="html-img"><img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g003.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g003.png" alt="Biomedicines 12 02216 g003" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g003.png" /></div> </div> <div class="html-fig-wrap" id="biomedicines-12-02216-f004"> <div class='html-fig_img'> <div class="html-figpopup html-figpopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f004"> <img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g004.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g004.png" alt="Biomedicines 12 02216 g004" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g004-550.jpg" /> <a class="html-expand html-figpopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f004"></a> </div> </div> <div class="html-fig_description"> <b>Figure 4.</b> The structural features of Kalata B1. (<b>A</b>) The amino acid sequence of Kalata B1. (<b>B</b>) An ensemble of averaged structure of Kalata B1 determined using NMR spectroscopy. The positions of the cysteines (labeled I through IV) and the corresponding disulfide linkages are shown. The * indicates a point of known cyclization between the glycine (C) and Asparagine (N) residues.  </div> </div> <div class="html-fig_show mfp-hide" id="fig_body_display_biomedicines-12-02216-f004"> <div class="html-caption"> <b>Figure 4.</b> The structural features of Kalata B1. (<b>A</b>) The amino acid sequence of Kalata B1. (<b>B</b>) An ensemble of averaged structure of Kalata B1 determined using NMR spectroscopy. The positions of the cysteines (labeled I through IV) and the corresponding disulfide linkages are shown. The * indicates a point of known cyclization between the glycine (C) and Asparagine (N) residues.</div> <div class="html-img"><img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g004.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g004.png" alt="Biomedicines 12 02216 g004" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g004.png" /></div> </div> <div class="html-fig-wrap" id="biomedicines-12-02216-f005"> <div class='html-fig_img'> <div class="html-figpopup html-figpopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f005"> <img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g005.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g005.png" alt="Biomedicines 12 02216 g005" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g005-550.jpg" /> <a class="html-expand html-figpopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f005"></a> </div> </div> <div class="html-fig_description"> <b>Figure 5.</b> Effect of natural and synthetic Kalata B1 on cell viability in SH-SY5Y, U-87MG, and T-98G cells cultures. The IC<sub>50</sub> values for the cyclotides were determined using MTT assay after incubation with cells at increasing concentrations (0.1, 1, 5, and 10 μM) as compared to the control. Panel (<b>A</b>): SH-SY5Y and U-87 MG cells, and panel (<b>B</b>) T-98G and U-87 MG cells. Cultures (6000 cells per well in a 96-well plate), were treated with the cyclotide indicated, for 72 h, then MTT measured. Cell viability was calculated as a percentage of control cultures. IC<sub>50</sub> values were calculated using the Best Fit method (See <a href="#biomedicines-12-02216-t001" class="html-table">Table 1</a>). Incubations were conducted in 96-well plates in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). The results are graphed in GraphPad Prism version 9 for iOS.  </div> </div> <div class="html-fig_show mfp-hide" id="fig_body_display_biomedicines-12-02216-f005"> <div class="html-caption"> <b>Figure 5.</b> Effect of natural and synthetic Kalata B1 on cell viability in SH-SY5Y, U-87MG, and T-98G cells cultures. The IC<sub>50</sub> values for the cyclotides were determined using MTT assay after incubation with cells at increasing concentrations (0.1, 1, 5, and 10 μM) as compared to the control. Panel (<b>A</b>): SH-SY5Y and U-87 MG cells, and panel (<b>B</b>) T-98G and U-87 MG cells. Cultures (6000 cells per well in a 96-well plate), were treated with the cyclotide indicated, for 72 h, then MTT measured. Cell viability was calculated as a percentage of control cultures. IC<sub>50</sub> values were calculated using the Best Fit method (See <a href="#biomedicines-12-02216-t001" class="html-table">Table 1</a>). Incubations were conducted in 96-well plates in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). The results are graphed in GraphPad Prism version 9 for iOS.</div> <div class="html-img"><img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g005.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g005.png" alt="Biomedicines 12 02216 g005" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g005.png" /></div> </div> <div class="html-fig-wrap" id="biomedicines-12-02216-f006"> <div class='html-fig_img'> <div class="html-figpopup html-figpopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f006"> <img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g006.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g006.png" alt="Biomedicines 12 02216 g006" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g006-550.jpg" /> <a class="html-expand html-figpopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f006"></a> </div> </div> <div class="html-fig_description"> <b>Figure 6.</b> Effect of increasing the concentration of naturally occurring cyclotides on cell viability in U-87 MG cells. IC<sub>50</sub> values for cyclotides were determined using the MTT assay after incubating U-87 MG cell cultures with increasing concentrations (0.1, 1.0, 5.0, and 10 μM) or with a control of DMSO only. Panel (<b>A</b>): CyO3, panel (<b>B</b>): CyO19, panel (<b>C</b>): Kalata B1, panel (<b>D</b>): Kalata B7, and panel (<b>E</b>), Vitri E. U-87 cells (6000 cells per well in a 96-well plate) were treated with cyclotides, as indicated, for 72 h, then MTT measured. Cell viability was calculated as a percentage of control cultures. IC<sub>50</sub> values were calculated using the Best Fit method (See <a href="#biomedicines-12-02216-t001" class="html-table">Table 1</a>). Incubations were conducted in 96-well plates in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). Results are graphed in GraphPad Prism version 9 for iOS.  </div> </div> <div class="html-fig_show mfp-hide" id="fig_body_display_biomedicines-12-02216-f006"> <div class="html-caption"> <b>Figure 6.</b> Effect of increasing the concentration of naturally occurring cyclotides on cell viability in U-87 MG cells. IC<sub>50</sub> values for cyclotides were determined using the MTT assay after incubating U-87 MG cell cultures with increasing concentrations (0.1, 1.0, 5.0, and 10 μM) or with a control of DMSO only. Panel (<b>A</b>): CyO3, panel (<b>B</b>): CyO19, panel (<b>C</b>): Kalata B1, panel (<b>D</b>): Kalata B7, and panel (<b>E</b>), Vitri E. U-87 cells (6000 cells per well in a 96-well plate) were treated with cyclotides, as indicated, for 72 h, then MTT measured. Cell viability was calculated as a percentage of control cultures. IC<sub>50</sub> values were calculated using the Best Fit method (See <a href="#biomedicines-12-02216-t001" class="html-table">Table 1</a>). Incubations were conducted in 96-well plates in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). Results are graphed in GraphPad Prism version 9 for iOS.</div> <div class="html-img"><img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g006.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g006.png" alt="Biomedicines 12 02216 g006" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g006.png" /></div> </div> <div class="html-fig-wrap" id="biomedicines-12-02216-f007"> <div class='html-fig_img'> <div class="html-figpopup html-figpopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f007"> <img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g007.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g007.png" alt="Biomedicines 12 02216 g007" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g007-550.jpg" /> <a class="html-expand html-figpopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f007"></a> </div> </div> <div class="html-fig_description"> <b>Figure 7.</b> Comparison of IC<sub>50</sub> values for natural and synthetic Kalata B1, as determined in SH-SY5Y, U-87 MG, and T-98G human cell cultures. Panel (<b>A</b>) includes SH-SY5Y and U-87 MG cells, panel (<b>B</b>) includes T-98G and U-87 MG cells, and panel (<b>C</b>) is natural Kalata B1 and synthetic Kalata B1 with concentrations of 0.1, 1, 5, and 10 μM as compared to the control. Cultures were plated in 96-well pates at 6000 cells/well, then treated with cyclotides, as indicated, for 72 h, then assayed using MTT. Cell viability was calculated as a percentage of control (where the control cells were treated with DMSO only). Incubations were conducted in 96-well plates in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). IC<sub>50</sub> values were calculated using the Best Fit method in GraphPad Prism version 8.0 (IC50 Best Fit). Results are graphed in GraphPad Prism version 8 for iOS.  </div> </div> <div class="html-fig_show mfp-hide" id="fig_body_display_biomedicines-12-02216-f007"> <div class="html-caption"> <b>Figure 7.</b> Comparison of IC<sub>50</sub> values for natural and synthetic Kalata B1, as determined in SH-SY5Y, U-87 MG, and T-98G human cell cultures. Panel (<b>A</b>) includes SH-SY5Y and U-87 MG cells, panel (<b>B</b>) includes T-98G and U-87 MG cells, and panel (<b>C</b>) is natural Kalata B1 and synthetic Kalata B1 with concentrations of 0.1, 1, 5, and 10 μM as compared to the control. Cultures were plated in 96-well pates at 6000 cells/well, then treated with cyclotides, as indicated, for 72 h, then assayed using MTT. Cell viability was calculated as a percentage of control (where the control cells were treated with DMSO only). Incubations were conducted in 96-well plates in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). IC<sub>50</sub> values were calculated using the Best Fit method in GraphPad Prism version 8.0 (IC50 Best Fit). Results are graphed in GraphPad Prism version 8 for iOS.</div> <div class="html-img"><img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g007.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g007.png" alt="Biomedicines 12 02216 g007" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g007.png" /></div> </div> <div class="html-fig-wrap" id="biomedicines-12-02216-f008"> <div class='html-fig_img'> <div class="html-figpopup html-figpopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f008"> <img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g008.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g008.png" alt="Biomedicines 12 02216 g008" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g008-550.jpg" /> <a class="html-expand html-figpopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f008"></a> </div> </div> <div class="html-fig_description"> <b>Figure 8.</b> Synthetic Kalata B1 increased TMZ-induced cell death in U-87 MG glioblastoma cultures. To determine if synthetic Kalata B1 (K) enhances TMZ (T)-induced cell death in U-87 MG cells, MTT was measured after incubating the cell cultures with concentrations of K (0.5, 1.0, 2.0, and 4.0 μM) and T (100, 200, 400, and 800 μM) both alone and when co-exposed to synthetic Kalata B1 and TMZ as illustrated in panels (<b>A</b>–<b>M</b>). The first dotted bars in each panel indicate the cells exposed to concentrations of synthetic Kalata B1 alone, while the second diagonal stripe bars indicate the cells exposed to TMZ alone. The third checkered bars indicate co-exposure treatments with synthetic Kalata B1 and TMZ. The cells were plated into 96-well plates (6000 cells/well) and treated with synthetic Kalata B, as indicated, for 72 h. Cell viability was measured using MTT and expressed as a percentage of control cultures (culture medium with DMSO only). Incubations were conducted in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). Results of one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons are graphed in GraphPad Prism version 9 for iOS with the following <span class='html-italic'>p</span>-values: * < 0.05, ** < 0.01, *** < 0.001, and **** < 0.0001. ns = not significant.  </div> </div> <div class="html-fig_show mfp-hide" id="fig_body_display_biomedicines-12-02216-f008"> <div class="html-caption"> <b>Figure 8.</b> Synthetic Kalata B1 increased TMZ-induced cell death in U-87 MG glioblastoma cultures. To determine if synthetic Kalata B1 (K) enhances TMZ (T)-induced cell death in U-87 MG cells, MTT was measured after incubating the cell cultures with concentrations of K (0.5, 1.0, 2.0, and 4.0 μM) and T (100, 200, 400, and 800 μM) both alone and when co-exposed to synthetic Kalata B1 and TMZ as illustrated in panels (<b>A</b>–<b>M</b>). The first dotted bars in each panel indicate the cells exposed to concentrations of synthetic Kalata B1 alone, while the second diagonal stripe bars indicate the cells exposed to TMZ alone. The third checkered bars indicate co-exposure treatments with synthetic Kalata B1 and TMZ. The cells were plated into 96-well plates (6000 cells/well) and treated with synthetic Kalata B, as indicated, for 72 h. Cell viability was measured using MTT and expressed as a percentage of control cultures (culture medium with DMSO only). Incubations were conducted in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). Data are expressed as mean ± standard error of the mean (SEM). Results of one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons are graphed in GraphPad Prism version 9 for iOS with the following <span class='html-italic'>p</span>-values: * < 0.05, ** < 0.01, *** < 0.001, and **** < 0.0001. ns = not significant.</div> <div class="html-img"><img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g008.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g008.png" alt="Biomedicines 12 02216 g008" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g008.png" /></div> </div> <div class="html-fig-wrap" id="biomedicines-12-02216-f009"> <div class='html-fig_img'> <div class="html-figpopup html-figpopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f009"> <img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g009.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g009.png" alt="Biomedicines 12 02216 g009" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g009-550.jpg" /> <a class="html-expand html-figpopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f009"></a> </div> </div> <div class="html-fig_description"> <b>Figure 9.</b> Synthetic Kalata B1 increased TMZ-induced cell death in T-98G glioblastoma cultures. To determine if Kalata B1 (K) enhances TMZ (T)-induced cell death in T-98G cells, MTT was measured after incubating cell cultures with synthetic Kalata B1 (0.25, 0.5, 1.0, and 2.0 μM) and TMZ (T 75, 150, 300, and 600 μM) both alone and when co-exposed as illustrated in panels (<b>A</b>–<b>I</b>). The first dotted bars in each panel indicate the cells exposed to synthetic Kalata B1 alone, while the second diagonal stripe bars indicate the cells exposed to TMZ alone. The third checkered bars indicate co-exposure treatments with synthetic Kalata B1 and TMZ. The cells were plated into 96-well plates (6000 cells/well) and treated with synthetic Kalata B1, as indicated, for 72 h. Cell viability was measured using MTT and expressed as a percentage of control cultures (culture medium + DMSO only). Incubations were conducted in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). The data are expressed as mean ± standard error of the mean (SEM). The results of one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons are graphed in GraphPad Prism version 9 for iOS, with the following <span class='html-italic'>p</span>-values: * < 0.05, ** < 0.01, *** < 0.001, and **** < 0.0001. ns = not significant.  </div> </div> <div class="html-fig_show mfp-hide" id="fig_body_display_biomedicines-12-02216-f009"> <div class="html-caption"> <b>Figure 9.</b> Synthetic Kalata B1 increased TMZ-induced cell death in T-98G glioblastoma cultures. To determine if Kalata B1 (K) enhances TMZ (T)-induced cell death in T-98G cells, MTT was measured after incubating cell cultures with synthetic Kalata B1 (0.25, 0.5, 1.0, and 2.0 μM) and TMZ (T 75, 150, 300, and 600 μM) both alone and when co-exposed as illustrated in panels (<b>A</b>–<b>I</b>). The first dotted bars in each panel indicate the cells exposed to synthetic Kalata B1 alone, while the second diagonal stripe bars indicate the cells exposed to TMZ alone. The third checkered bars indicate co-exposure treatments with synthetic Kalata B1 and TMZ. The cells were plated into 96-well plates (6000 cells/well) and treated with synthetic Kalata B1, as indicated, for 72 h. Cell viability was measured using MTT and expressed as a percentage of control cultures (culture medium + DMSO only). Incubations were conducted in triplicate wells (technical replicates) and as three independent incubations (biological replicates, n = 3). The data are expressed as mean ± standard error of the mean (SEM). The results of one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons are graphed in GraphPad Prism version 9 for iOS, with the following <span class='html-italic'>p</span>-values: * < 0.05, ** < 0.01, *** < 0.001, and **** < 0.0001. ns = not significant.</div> <div class="html-img"><img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g009.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g009.png" alt="Biomedicines 12 02216 g009" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g009.png" /></div> </div> <div class="html-fig-wrap" id="biomedicines-12-02216-f010"> <div class='html-fig_img'> <div class="html-figpopup html-figpopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f010"> <img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g010.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g010.png" alt="Biomedicines 12 02216 g010" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g010-550.jpg" /> <a class="html-expand html-figpopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f010"></a> </div> </div> <div class="html-fig_description"> <b>Figure 10.</b> Phase contrast microscopy of U-87 GM cultures treated with Kalata B1 and temozolomide. Panels are as follows: (<b>A</b>) no treatment/control; (<b>B</b>) 150 µM temozolomide (TMZ); (<b>C</b>) synthetic Kalata B1 0.5 µM; and (<b>D</b>) 150 µM TMZ + 0.5 µM synthetic Kalata B1 for 72 h. Imaged on an Olympus IX83 inverted microscope equipped with an Olympus XM10 camera and acquired in cellSens standard. Images were adjusted for exposure in Adobe Photoshop 23.4.2 using “Levels” and cropped for consistency and to enable ease of comparison. U-87 GM cells were plated at 6000 cells/well in a 96-well plate and allowed to adhere overnight prior to treatment. Images are representative of triplicate incubations. The yellow bar in the lower right-hand corner of the image is 50 µm. 20× magnification.  </div> </div> <div class="html-fig_show mfp-hide" id="fig_body_display_biomedicines-12-02216-f010"> <div class="html-caption"> <b>Figure 10.</b> Phase contrast microscopy of U-87 GM cultures treated with Kalata B1 and temozolomide. Panels are as follows: (<b>A</b>) no treatment/control; (<b>B</b>) 150 µM temozolomide (TMZ); (<b>C</b>) synthetic Kalata B1 0.5 µM; and (<b>D</b>) 150 µM TMZ + 0.5 µM synthetic Kalata B1 for 72 h. Imaged on an Olympus IX83 inverted microscope equipped with an Olympus XM10 camera and acquired in cellSens standard. Images were adjusted for exposure in Adobe Photoshop 23.4.2 using “Levels” and cropped for consistency and to enable ease of comparison. U-87 GM cells were plated at 6000 cells/well in a 96-well plate and allowed to adhere overnight prior to treatment. Images are representative of triplicate incubations. The yellow bar in the lower right-hand corner of the image is 50 µm. 20× magnification.</div> <div class="html-img"><img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g010.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g010.png" alt="Biomedicines 12 02216 g010" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g010.png" /></div> </div> <div class="html-fig-wrap" id="biomedicines-12-02216-f011"> <div class='html-fig_img'> <div class="html-figpopup html-figpopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f011"> <img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g011.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g011.png" alt="Biomedicines 12 02216 g011" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g011-550.jpg" /> <a class="html-expand html-figpopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#fig_body_display_biomedicines-12-02216-f011"></a> </div> </div> <div class="html-fig_description"> <b>Figure 11.</b> Stability of synthetic Kalata B1 and temozolomide (TMZ) in human serum over 24 h. Percentage of Kalata B1 and TMZ remaining upon incubation with human serum. The portion of serum-treated peptide remaining was determined by calculating the area of the respective elution peak in UPLC-MS and comparing it with the control without incubation. Data points are means ± SEM.  </div> </div> <div class="html-fig_show mfp-hide" id="fig_body_display_biomedicines-12-02216-f011"> <div class="html-caption"> <b>Figure 11.</b> Stability of synthetic Kalata B1 and temozolomide (TMZ) in human serum over 24 h. Percentage of Kalata B1 and TMZ remaining upon incubation with human serum. The portion of serum-treated peptide remaining was determined by calculating the area of the respective elution peak in UPLC-MS and comparing it with the control without incubation. Data points are means ± SEM.</div> <div class="html-img"><img data-large="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g011.png" data-original="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g011.png" alt="Biomedicines 12 02216 g011" data-lsrc="/biomedicines/biomedicines-12-02216/article_deploy/html/images/biomedicines-12-02216-g011.png" /></div> </div> <div class="html-table-wrap" id="biomedicines-12-02216-t001"> <div class="html-table_wrap_td"> <div class="html-tablepopup html-tablepopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href='#table_body_display_biomedicines-12-02216-t001'> <img data-lsrc="https://pub.mdpi-res.com/img/table.png" /> <a class="html-expand html-tablepopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#table_body_display_biomedicines-12-02216-t001"></a> </div> </div> <div class="html-table_wrap_discription"> <b>Table 1.</b> MS/MS analysis of peptide 1 using orbitrap mass spectrometry. </div> </div> <div class="html-table_show mfp-hide " id="table_body_display_biomedicines-12-02216-t001"> <div class="html-caption"><b>Table 1.</b> MS/MS analysis of peptide 1 using orbitrap mass spectrometry.</div> <table > <thead ><tr ><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' >Sequence</th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' ><span class='html-italic'>m</span>/<span class='html-italic'>z</span></th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' >Observed Mass Difference</th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' >Expected Mass Difference</th></tr></thead><tbody ><tr ><td align='center' valign='middle' class='html-align-center' >N</td><td align='center' valign='middle' class='html-align-center' >845.3799</td><td align='center' valign='middle' class='html-align-center' > </td><td align='center' valign='middle' class='html-align-center' > </td></tr><tr ><td align='center' valign='middle' class='html-align-center' >G</td><td align='center' valign='middle' class='html-align-center' >731.33765</td><td align='center' valign='middle' class='html-align-center' >114.04236</td><td align='center' valign='middle' class='html-align-center' >114.04293</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >L</td><td align='center' valign='middle' class='html-align-center' >674.31616</td><td align='center' valign='middle' class='html-align-center' >57.02148</td><td align='center' valign='middle' class='html-align-center' >57.02146</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >P</td><td align='center' valign='middle' class='html-align-center' >561.23236</td><td align='center' valign='middle' class='html-align-center' >113.0838</td><td align='center' valign='middle' class='html-align-center' >113.08406</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >V</td><td align='center' valign='middle' class='html-align-center' >464.17984</td><td align='center' valign='middle' class='html-align-center' >97.05252</td><td align='center' valign='middle' class='html-align-center' >97.05276</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >C</td><td align='center' valign='middle' class='html-align-center' >365.111172</td><td align='center' valign='middle' class='html-align-center' >99.06812</td><td align='center' valign='middle' class='html-align-center' >99.06841</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >G</td><td align='center' valign='middle' class='html-align-center' >205.0815</td><td align='center' valign='middle' class='html-align-center' >160.03023</td><td align='center' valign='middle' class='html-align-center' >160.03059</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >E</td><td align='center' valign='middle' class='html-align-center' >148.06015</td><td align='center' valign='middle' class='html-align-center' >57.02135</td><td align='center' valign='middle' class='html-align-center' >57.02146</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >148.060984</td></tr></tbody> </table> </div> <div class="html-table-wrap" id="biomedicines-12-02216-t002"> <div class="html-table_wrap_td"> <div class="html-tablepopup html-tablepopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href='#table_body_display_biomedicines-12-02216-t002'> <img data-lsrc="https://pub.mdpi-res.com/img/table.png" /> <a class="html-expand html-tablepopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#table_body_display_biomedicines-12-02216-t002"></a> </div> </div> <div class="html-table_wrap_discription"> <b>Table 2.</b> MS/MS analysis of peptide 2 using orbitrap mass spectrometry. </div> </div> <div class="html-table_show mfp-hide " id="table_body_display_biomedicines-12-02216-t002"> <div class="html-caption"><b>Table 2.</b> MS/MS analysis of peptide 2 using orbitrap mass spectrometry.</div> <table > <thead ><tr ><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' >Sequence</th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' ><span class='html-italic'>m</span>/<span class='html-italic'>z</span></th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' >Observed Mass Difference</th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' >Expected Mass Difference</th></tr></thead><tbody ><tr ><td align='center' valign='middle' class='html-align-center' >R</td><td align='center' valign='middle' class='html-align-center' > </td><td align='center' valign='middle' class='html-align-center' > </td><td align='center' valign='middle' class='html-align-center' >175.119501</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >T</td><td align='center' valign='middle' class='html-align-center' >175.1182</td><td align='center' valign='middle' class='html-align-center' >101.04756</td><td align='center' valign='middle' class='html-align-center' >101.04768</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >C</td><td align='center' valign='middle' class='html-align-center' >276.16638</td><td align='center' valign='middle' class='html-align-center' >160.03055</td><td align='center' valign='middle' class='html-align-center' >160.03059</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >V</td><td align='center' valign='middle' class='html-align-center' >436.19693</td><td align='center' valign='middle' class='html-align-center' >99.06827</td><td align='center' valign='middle' class='html-align-center' >99.06841</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >P</td><td align='center' valign='middle' class='html-align-center' >535.2652</td><td align='center' valign='middle' class='html-align-center' >97.05267</td><td align='center' valign='middle' class='html-align-center' >97.05276</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >W</td><td align='center' valign='middle' class='html-align-center' >632.31787</td><td align='center' valign='middle' class='html-align-center' >186.07916</td><td align='center' valign='middle' class='html-align-center' >186.07931</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >S</td><td align='center' valign='middle' class='html-align-center' >818.39703</td><td align='center' valign='middle' class='html-align-center' >87.03192</td><td align='center' valign='middle' class='html-align-center' >87.03203</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >C</td><td align='center' valign='middle' class='html-align-center' >905.42896</td><td align='center' valign='middle' class='html-align-center' >160.03052</td><td align='center' valign='middle' class='html-align-center' >160.03059</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >T</td><td align='center' valign='middle' class='html-align-center' >1065.45947</td><td align='center' valign='middle' class='html-align-center' >101.04785</td><td align='center' valign='middle' class='html-align-center' >101.04768</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >C</td><td align='center' valign='middle' class='html-align-center' >1166.50732</td><td align='center' valign='middle' class='html-align-center' >160.03113</td><td align='center' valign='middle' class='html-align-center' >160.03059</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >G</td><td align='center' valign='middle' class='html-align-center' >1326.53845</td><td align='center' valign='middle' class='html-align-center' >57.02136</td><td align='center' valign='middle' class='html-align-center' >57.02146</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >P</td><td align='center' valign='middle' class='html-align-center' >1383.55981</td><td align='center' valign='middle' class='html-align-center' >97.05212</td><td align='center' valign='middle' class='html-align-center' >97.05276</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >T</td><td align='center' valign='middle' class='html-align-center' >1480.61194</td><td align='center' valign='middle' class='html-align-center' >101.04785</td><td align='center' valign='middle' class='html-align-center' >101.04768</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >N</td><td align='center' valign='middle' class='html-align-center' >1581.65979</td><td align='center' valign='middle' class='html-align-center' >114.04272</td><td align='center' valign='middle' class='html-align-center' >114.04293</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >C</td><td align='center' valign='middle' class='html-align-center' >1695.70251</td><td align='center' valign='middle' class='html-align-center' >160.03076</td><td align='center' valign='middle' class='html-align-center' >160.03059</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >T</td><td align='center' valign='middle' class='html-align-center' >1855.73328</td><td align='center' valign='middle' class='html-align-center' >57.02209</td><td align='center' valign='middle' class='html-align-center' >57.02146</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >G</td><td align='center' valign='middle' class='html-align-center' >1956.7821</td><td align='center' valign='middle' class='html-align-center' >57.02124</td><td align='center' valign='middle' class='html-align-center' >57.02146</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >G</td><td align='center' valign='middle' class='html-align-center' >2013.8042</td><td align='center' valign='middle' class='html-align-center' >99.06763</td><td align='center' valign='middle' class='html-align-center' >99.06841</td></tr><tr ><td align='center' valign='middle' class='html-align-center' >V</td><td align='center' valign='middle' class='html-align-center' >2070.82544</td><td align='center' valign='middle' class='html-align-center' >ND</td><td align='center' valign='middle' class='html-align-center' > </td></tr><tr ><td align='center' valign='middle' class='html-align-center' >C</td><td align='center' valign='middle' class='html-align-center' >2169.89307</td><td align='center' valign='middle' class='html-align-center' >ND</td><td align='center' valign='middle' class='html-align-center' > </td></tr><tr ><td align='center' valign='middle' class='html-align-center' >T</td><td align='center' valign='middle' class='html-align-center' >ND</td><td align='center' valign='middle' class='html-align-center' > </td><td align='center' valign='middle' class='html-align-center' > </td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >ND</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td></tr></tbody> </table> <div class='html-table_foot html-p'><div class='html-p' style='text-indent:0em;'><span class='html-fn-content'>ND, not detected.</span></div><div style='clear:both;'></div></div> </div> <div class="html-table-wrap" id="biomedicines-12-02216-t003"> <div class="html-table_wrap_td"> <div class="html-tablepopup html-tablepopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href='#table_body_display_biomedicines-12-02216-t003'> <img data-lsrc="https://pub.mdpi-res.com/img/table.png" /> <a class="html-expand html-tablepopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#table_body_display_biomedicines-12-02216-t003"></a> </div> </div> <div class="html-table_wrap_discription"> <b>Table 3.</b> Chemical shifts of Kalata B1. </div> </div> <div class="html-table_show mfp-hide " id="table_body_display_biomedicines-12-02216-t003"> <div class="html-caption"><b>Table 3.</b> Chemical shifts of Kalata B1.</div> <table > <thead ><tr ><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' >Residues</th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' > </th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' > </th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' > </th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' > </th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' > </th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' > </th><th align='center' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-center' > </th></tr><tr ><th align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </th><th align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >HN</th><th align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Hα</th><th align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Hβ</th><th align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Hγ</th><th align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Hδ</th><th align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Others</th><th align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3JNH-CaH (Hz)</th></tr></thead><tbody ><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Gly1</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.66</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.15, 3.50</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Leu 2</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >7.67</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.96</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.84, 1.39</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.62</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >0.89, 0.82</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >10.13</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Pro 3</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.98</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >2.37, 1.63</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >2.06, 1.96</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.70, 3.67</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Val 4</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.04</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.57</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >2.50</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >0.79, 0.75</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.5</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Cys 5</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >7.90</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.38</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.26, 2.91</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.83</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Gly 6</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.43</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.66</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Glu 7</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >7.08</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.71</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.93, 1.78</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >2.45</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.11</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Thr 8</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.36</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.47</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.36</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.05</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.25</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Cys 9</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.25</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.85</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.10, 2.83</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >11.2</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Val 10</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.43</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.76</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.95</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >0.94, 0.86</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.2</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Gly 11</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.59</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.17, 3.75</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Gly 12</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.13</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.32, 3.95</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Thr 13</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >7.76</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.62</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.01</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.06</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Cys 14</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.60</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.61</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >2.95, 2.68</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.77</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Asn 15</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.33</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.61</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >2.94, 2.70</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.98</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Thr 16</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.27</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.41</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.12</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.24</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.36</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Pro 17</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.18</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >2.23, 1.82</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >2.06, 1.92</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.06, 3.62</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Gly 18</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.68</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.19, 3.60</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Cys 19</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >7.60</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >5.23</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.73, 2.53</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >10.28</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Thr 20</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.42</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.44</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.95</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.04</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.17</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Cys 21</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.85</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.5</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.03, 2.73</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.62</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Ser 22</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.87</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.68</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.76</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.62</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Trp 23</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >7.88</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.98</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.16</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >7.21</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >10.31 (HNε), 7.34 (Hε3), 7.13 (HH2)</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >7.42 (HZ2), 7.02 (HZ3)</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Pro 24</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.33</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.59 -0.30</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.27, 1.19</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.15, 3.11</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Val 25</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.18</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.12</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.83</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >0.76, 0.73</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.01</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Cys 26</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >7.63</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >5.02</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.13, 2.66</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.37</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Thr 27</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.77</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.98</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.62</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >0.79</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.74</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Arg 28</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >8.62</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.67</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.58</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >1.34</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.07</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >6.86 (Hε)</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >10.6</td></tr><tr ><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >Asn 29</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.45</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >4.31</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >3.00, 2.75</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' > </td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >-</td><td align='center' valign='middle' style='border-bottom:solid thin' class='html-align-center' >9.87</td></tr></tbody> </table> </div> <div class="html-table-wrap" id="biomedicines-12-02216-t004"> <div class="html-table_wrap_td"> <div class="html-tablepopup html-tablepopup-link" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href='#table_body_display_biomedicines-12-02216-t004'> <img data-lsrc="https://pub.mdpi-res.com/img/table.png" /> <a class="html-expand html-tablepopup" data-counterslinkmanual = "https://www.mdpi.com/2227-9059/12/10/2216/display" href="#table_body_display_biomedicines-12-02216-t004"></a> </div> </div> <div class="html-table_wrap_discription"> <b>Table 4.</b> Toxicity and IC<sub>50</sub> values of five cyclotides and TMZ in SH-SY5Y, U-87 MG, and T-98G cell cultures. </div> </div> <div class="html-table_show mfp-hide " id="table_body_display_biomedicines-12-02216-t004"> <div class="html-caption"><b>Table 4.</b> Toxicity and IC<sub>50</sub> values of five cyclotides and TMZ in SH-SY5Y, U-87 MG, and T-98G cell cultures.</div> <table > <thead ><tr ><th align='left' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-left' >Cell Type</th><th align='left' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-left' >Compound</th><th align='left' valign='middle' style='border-top:solid thin;border-bottom:solid thin' class='html-align-left' >IC<sub>50</sub> (μM) ± S.D.</th></tr></thead><tbody ><tr ><td rowspan='8' align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >U-87 MG</td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >CyO3</td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >2.6 ± 0.05</td></tr><tr ><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >CyO19</td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >2.4 ± 0.07</td></tr><tr ><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >Kalata B1 <sup>a</sup></td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >7.9 ± 1.66</td></tr><tr ><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >Kalata B1 (synthetic)</td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >5.4 ± 1.06</td></tr><tr ><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >Kalata B7</td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >10.1 ± 1.74</td></tr><tr ><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >Vitri E</td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >21.1 ± 2.90</td></tr><tr ><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >TMZ <sup>a</sup></td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >2898.0 ± 6.00</td></tr><tr ><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >TMZ</td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >1617.0 ± 5.56</td></tr><tr ><td rowspan='2' align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >T-98G</td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >Kalata B1 (synthetic)</td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >2.6 ± 0.24</td></tr><tr ><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >TMZ</td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >1160.0 ± 5.31</td></tr><tr ><td rowspan='2' align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >SH-SY5Y</td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >Kalata B1 <sup>a</sup></td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >5.6 ± 1.71</td></tr><tr ><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >TMZ <sup>a</sup></td><td align='left' valign='middle' style='border-bottom:solid thin' class='html-align-left' >393.0 ± 4.50</td></tr></tbody> </table> <div class='html-table_foot html-p'><div class='html-p' style='text-indent:0em;'><span class='html-fn-content'><sup>a</sup> indicates previously published data [<a href="#B9-biomedicines-12-02216" class="html-bibr">9</a>].</span></div><div style='clear:both;'></div></div> </div> </section><section class='html-fn_group'><table><tr id=''><td></td><td><div class='html-p'><b>Disclaimer/Publisher’s Note:</b> The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.</div></td></tr></table></section> <section id="html-copyright"><br>© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (<a href="https://creativecommons.org/licenses/by/4.0/" target="_blank" rel="noopener noreferrer">https://creativecommons.org/licenses/by/4.0/</a>).</section> </div> </div> <div class="additional-content"> <h2><a name="cite"></a>Share and Cite</h2> <div class="social-media-links" style="text-align: left;"> <a href="/cdn-cgi/l/email-protection#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" title="Email"> <i class="fa fa-envelope-square" style="font-size: 30px;"></i> </a> <a href="https://twitter.com/intent/tweet?text=Kalata+B1+Enhances+Temozolomide+Toxicity+to+Glioblastoma+Cells&hashtags=mdpibiomedicines&url=https%3A%2F%2Fwww.mdpi.com%2F2976274&via=Biomed_MDPI" onclick="windowOpen(this.href,600,800); return false" target="_blank" rel="noopener noreferrer"> <i class="fa fa-twitter-x-square" style="font-size: 30px;"></i> </a> <a href=" http://www.linkedin.com/shareArticle?mini=true&url=https%3A%2F%2Fwww.mdpi.com%2F2976274&title=Kalata%20B1%20Enhances%20Temozolomide%20Toxicity%20to%20Glioblastoma%20Cells%26source%3Dhttps%3A%2F%2Fwww.mdpi.com%26summary%3DGlioblastoma%20%28GBM%29%20is%20the%20most%20aggressive%20cancer%20originating%20in%20the%20brain%2C%20but%20unfortunately%20combination%20treatments%20with%20resection%2C%20radiation%2C%20and%20chemotherapy%20are%20relatively%20ineffective.%20Therefore%2C%20novel%20methods%20of%20adjuvant%20therapy%20are%20critically%20%5B...%5D" onclick="windowOpen(this.href,600,800); return false" title="LinkedIn" target="_blank" rel="noopener noreferrer"> <i class="fa fa-linkedin-square" style="font-size: 30px;"></i> </a> <a href="https://www.facebook.com/sharer.php?u=https://www.mdpi.com/2976274" title="facebook" target="_blank" rel="noopener noreferrer"> <i class="fa fa-facebook-square" style="font-size: 30px;"></i> </a> <a href="javascript:void(0);" title="Wechat" data-reveal-id="weixin-share-modal"> <i class="fa fa-weixin-square" style="font-size: 26px;"></i> </a> <a href="http://www.reddit.com/submit?url=https://www.mdpi.com/2976274" title="Reddit" target="_blank" rel="noopener noreferrer"> <i class="fa fa-reddit-square" style="font-size: 30px;"></i> </a> <a href="http://www.mendeley.com/import/?url=https://www.mdpi.com/2976274" title="Mendeley" target="_blank" rel="noopener noreferrer"> <i class="fa fa-mendeley-square" style="font-size: 30px;"></i> </a> </div> <div class="in-tab" style="padding-top: 0px!important; margin-top: 15px;"> <div><b>MDPI and ACS Style</b></div> <p> Gerlach, S.L.; Metcalf, J.S.; Dunlop, R.A.; Banack, S.A.; Her, C.; Krishnan, V.V.; Göransson, U.; Gunasekera, S.; Slazak, B.; Cox, P.A. Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells. <em>Biomedicines</em> <b>2024</b>, <em>12</em>, 2216. https://doi.org/10.3390/biomedicines12102216 </p> <div style="display: block"> <b>AMA Style</b><br> <p> Gerlach SL, Metcalf JS, Dunlop RA, Banack SA, Her C, Krishnan VV, Göransson U, Gunasekera S, Slazak B, Cox PA. Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells. <em>Biomedicines</em>. 2024; 12(10):2216. https://doi.org/10.3390/biomedicines12102216 </p> <b>Chicago/Turabian Style</b><br> <p> Gerlach, Samantha L., James S. Metcalf, Rachael A. Dunlop, Sandra Anne Banack, Cheenou Her, Viswanathan V. Krishnan, Ulf Göransson, Sunithi Gunasekera, Blazej Slazak, and Paul Alan Cox. 2024. "Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells" <em>Biomedicines</em> 12, no. 10: 2216. https://doi.org/10.3390/biomedicines12102216 </p> <b>APA Style</b><br> <p> Gerlach, S. L., Metcalf, J. S., Dunlop, R. A., Banack, S. A., Her, C., Krishnan, V. V., Göransson, U., Gunasekera, S., Slazak, B., & Cox, P. A. (2024). Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells. <em>Biomedicines</em>, <em>12</em>(10), 2216. https://doi.org/10.3390/biomedicines12102216 </p> </div> </div> <div class="info-box no-margin"> Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details <a target="_blank" href="https://www.mdpi.com/about/announcements/784">here</a>. </div> <h2><a name="metrics"></a>Article Metrics</h2> <div class="row"> <div class="small-12 columns"> <div id="loaded_cite_count" style="display:none">No</div> <div id="framed_div_cited_count" class="in-tab" style="display: none; overflow: auto;"></div> <div id="loaded" style="display:none">No</div> <div id="framed_div" class="in-tab" style="display: none; margin-top: 10px;"></div> </div> <div class="small-12 columns"> <div id="article_stats_div" style="display: none; margin-bottom: 1em;"> <h3>Article Access Statistics</h3> <div id="article_stats_swf" ></div> For more information on the journal statistics, click <a href="/journal/biomedicines/stats">here</a>. <div class="info-box"> Multiple requests from the same IP address are counted as one view. </div> </div> </div> </div> </div> </div> </article> </div> <div id="supplementaryModal" class="reveal-modal reveal-modal-new" data-reveal aria-labelledby="Captcha" aria-hidden="true" role="dialog"> <div class="row"> <div class="large-12 medium-12 small-12 columns"> <h2>Supplementary Material</h2> <div class="custom-accordion-for-small-screen-content show-for-medium-up"> <div class="in-tab"> <ul style="margin:0; list-style: none; overflow: auto;"> <li><a name="supplementary_file1"></a><b>Supplementary File 1:</b> <p> <a href="/2227-9059/12/10/2216/s1?version=1727511843"> ZIP-Document </a> (ZIP, 1040 KiB) </p> </li> </ul> </div> </div> </div> </div> <a class="close-reveal-modal" aria-label="Close"> <i class="material-icons">clear</i> </a> </div> </div></div> <div class="webpymol-controls webpymol-controls-template" style="margin-top: 10px; display: none;"> <a class="bzoom">Zoom</a> <span style="display: inline-block; margin-left: 5px; margin-right: 5px;">|</span> <a class="borient"> Orient </a> <span style="display: inline-block; margin-left: 5px; margin-right: 5px;">|</span> <a class="blines"> As Lines </a> <span style="display: inline-block; margin-left: 5px; margin-right: 5px;">|</span> <a class="bsticks"> As Sticks </a> <span style="display: inline-block; margin-left: 5px; margin-right: 5px;">|</span> <a class="bcartoon"> As Cartoon </a> <span style="display: inline-block; margin-left: 5px; margin-right: 5px;">|</span> <a class="bsurface"> As Surface </a> <span style="display: inline-block; margin-left: 5px; margin-right: 5px;">|</span> <a class="bprevscene">Previous Scene</a> <span style="display: inline-block; margin-left: 5px; margin-right: 5px;">|</span> <a class="bnextscene">Next Scene</a> </div> <div id="scifeed-modal" class="reveal-modal reveal-modal-new" data-reveal aria-labelledby="modalTitle" aria-hidden="true" role="dialog"> </div> <div id="recommended-articles-modal" class="reveal-modal reveal-modal-new" data-reveal aria-labelledby="modalTitle" aria-hidden="true" role="dialog"> </div> <div id="author-biographies-modal" class="reveal-modal reveal-modal-new" data-reveal aria-labelledby="modalTitle" aria-hidden="true" role="dialog"> </div> <div id="cite-modal" class="reveal-modal reveal-modal-new" data-reveal aria-labelledby="Captcha" aria-hidden="true" role="dialog"> <div class="row"> <div class="small-12 columns"> <h2 style="margin: 0;">Cite</h2> </div> <div class="small-12 columns">  <form style="margin:0; padding:0; display:inline;" name="export-bibtex" method="POST" action="/export"> <input type="hidden" name="articles_ids[]" value="1488137"> <input type="hidden" name="export_format_top" value="bibtex"> <input type="hidden" name="export_submit_top" value=""> </form>  <form style="margin:0; padding:0; display:inline;" name="export-endnote" method="POST" action="/export"> <input type="hidden" name="articles_ids[]" value="1488137"> <input type="hidden" name="export_format_top" value="endnote_no_abstract"> <input type="hidden" name="export_submit_top" value=""> </form>  <form style="margin:0; padding:0; display:inline;" name="export-ris" method="POST" action="/export"> <input type="hidden" name="articles_ids[]" value="1488137"> <input type="hidden" name="export_format_top" value="ris"> <input type="hidden" name="export_submit_top" value=""> </form> <div> Export citation file: <a href="javascript:window.document.forms['export-bibtex'].submit()">BibTeX</a> | <a href="javascript:window.document.forms['export-endnote'].submit()">EndNote</a> | <a href="javascript:window.document.forms['export-ris'].submit()">RIS</a> </div> </div> <div class="small-12 columns"> <div class="in-tab"> <div><b>MDPI and ACS Style</b></div> <p> Gerlach, S.L.; Metcalf, J.S.; Dunlop, R.A.; Banack, S.A.; Her, C.; Krishnan, V.V.; Göransson, U.; Gunasekera, S.; Slazak, B.; Cox, P.A. Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells. <em>Biomedicines</em> <b>2024</b>, <em>12</em>, 2216. https://doi.org/10.3390/biomedicines12102216 </p> <div style="display: block"> <b>AMA Style</b><br> <p> Gerlach SL, Metcalf JS, Dunlop RA, Banack SA, Her C, Krishnan VV, Göransson U, Gunasekera S, Slazak B, Cox PA. Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells. <em>Biomedicines</em>. 2024; 12(10):2216. https://doi.org/10.3390/biomedicines12102216 </p> <b>Chicago/Turabian Style</b><br> <p> Gerlach, Samantha L., James S. Metcalf, Rachael A. Dunlop, Sandra Anne Banack, Cheenou Her, Viswanathan V. Krishnan, Ulf Göransson, Sunithi Gunasekera, Blazej Slazak, and Paul Alan Cox. 2024. "Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells" <em>Biomedicines</em> 12, no. 10: 2216. https://doi.org/10.3390/biomedicines12102216 </p> <b>APA Style</b><br> <p> Gerlach, S. L., Metcalf, J. S., Dunlop, R. A., Banack, S. A., Her, C., Krishnan, V. V., Göransson, U., Gunasekera, S., Slazak, B., & Cox, P. A. (2024). Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells. <em>Biomedicines</em>, <em>12</em>(10), 2216. https://doi.org/10.3390/biomedicines12102216 </p> </div> </div> <div class="info-box no-margin"> Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details <a target="_blank" href="https://www.mdpi.com/about/announcements/784">here</a>. </div> </div> </div> <a class="close-reveal-modal" aria-label="Close"> <i class="material-icons">clear</i> </a> </div> </div> </div> </div> </div> </section> <div id="footer"> <div class="journal-info"> <span> <em><a class="Var_JournalInfo" href="/journal/biomedicines">Biomedicines</a></em>, EISSN 2227-9059, Published by MDPI </span> <div class="large-right"> <span> <a href="/rss/journal/biomedicines" class="rss-link">RSS</a> </span> <span> <a href="/journal/biomedicines/toc-alert">Content Alert</a> </span> </div> </div> <div class="row full-width footer-links" data-equalizer="footer" data-equalizer-mq="small"> <div class="large-2 large-push-4 medium-3 small-6 columns" data-equalizer-watch="footer"> <h3> Further Information </h3> <a href="/apc"> Article Processing Charges </a> <a href="/about/payment"> Pay an Invoice </a> <a href="/openaccess"> Open Access Policy </a> <a href="/about/contact"> Contact MDPI </a> <a href="https://careers.mdpi.com" target="_blank" rel="noopener noreferrer"> Jobs at MDPI </a> </div> <div class="large-2 large-push-4 medium-3 small-6 columns" data-equalizer-watch="footer"> <h3> Guidelines </h3> <a href="/authors"> For Authors </a> <a href="/reviewers"> For Reviewers </a> <a href="/editors"> For Editors </a> <a href="/librarians"> For Librarians </a> <a href="/publishing_services"> For Publishers </a> <a href="/societies"> For Societies </a> <a href="/conference_organizers"> For Conference Organizers </a> </div> <div class="large-2 large-push-4 medium-3 small-6 columns"> <h3> MDPI Initiatives </h3> <a href="https://sciforum.net" target="_blank" rel="noopener noreferrer"> Sciforum </a> <a href="https://www.mdpi.com/books" target="_blank" rel="noopener noreferrer"> MDPI Books </a> <a href="https://www.preprints.org" target="_blank" rel="noopener noreferrer"> Preprints.org </a> <a href="https://www.scilit.net" target="_blank" rel="noopener noreferrer"> Scilit </a> <a href="https://sciprofiles.com?utm_source=mpdi.com&utm_medium=bottom_menu&utm_campaign=initiative" target="_blank" rel="noopener noreferrer"> SciProfiles </a> <a href="https://encyclopedia.pub" target="_blank" rel="noopener noreferrer"> Encyclopedia </a> <a href="https://jams.pub" target="_blank" rel="noopener noreferrer"> JAMS </a> <a href="/about/proceedings"> Proceedings Series </a> </div> <div class="large-2 large-push-4 medium-3 small-6 right-border-large-without columns UA_FooterFollowMDPI"> <h3> Follow MDPI </h3> <a href="https://www.linkedin.com/company/mdpi" target="_blank" rel="noopener noreferrer"> LinkedIn </a> <a href="https://www.facebook.com/MDPIOpenAccessPublishing" target="_blank" rel="noopener noreferrer"> Facebook </a> <a href="https://twitter.com/MDPIOpenAccess" target="_blank" rel="noopener noreferrer"> Twitter </a> </div> <div id="footer-subscribe" class="large-4 large-pull-8 medium-12 small-12 left-border-large columns"> <div class="footer-subscribe__container"> <img class="show-for-large-up" src="https://pub.mdpi-res.com/img/design/mdpi-pub-logo-white-small.png?71d18e5f805839ab?1732615622" alt="MDPI" title="MDPI Open Access Journals" style="height: 50px; margin-bottom: 10px;"> <form id="newsletter" method="POST" action="/subscribe"> <p> Subscribe to receive issue release notifications and newsletters from MDPI journals </p> <select multiple id="newsletter-journal" class="foundation-select" name="journals[]"> <option value="acoustics">Acoustics</option> <option value="amh">Acta Microbiologica Hellenica</option> <option value="actuators">Actuators</option> <option value="admsci">Administrative Sciences</option> <option value="adolescents">Adolescents</option> <option value="arm">Advances in Respiratory Medicine</option> <option value="aerobiology">Aerobiology</option> <option value="aerospace">Aerospace</option> <option value="agriculture">Agriculture</option> <option value="agriengineering">AgriEngineering</option> <option value="agrochemicals">Agrochemicals</option> <option value="agronomy">Agronomy</option> <option value="ai">AI</option> <option value="air">Air</option> <option value="algorithms">Algorithms</option> <option value="allergies">Allergies</option> <option value="alloys">Alloys</option> <option value="analytica">Analytica</option> <option value="analytics">Analytics</option> <option value="anatomia">Anatomia</option> <option value="anesthres">Anesthesia Research</option> <option value="animals">Animals</option> <option value="antibiotics">Antibiotics</option> <option value="antibodies">Antibodies</option> <option value="antioxidants">Antioxidants</option> <option value="applbiosci">Applied Biosciences</option> <option value="applmech">Applied Mechanics</option> <option value="applmicrobiol">Applied Microbiology</option> <option value="applnano">Applied Nano</option> <option value="applsci">Applied Sciences</option> <option value="asi">Applied System Innovation</option> <option value="appliedchem">AppliedChem</option> <option value="appliedmath">AppliedMath</option> <option value="aquacj">Aquaculture Journal</option> <option value="architecture">Architecture</option> <option value="arthropoda">Arthropoda</option> <option value="arts">Arts</option> <option value="astronomy">Astronomy</option> <option value="atmosphere">Atmosphere</option> <option value="atoms">Atoms</option> <option value="audiolres">Audiology Research</option> <option value="automation">Automation</option> <option value="axioms">Axioms</option> <option value="bacteria">Bacteria</option> <option value="batteries">Batteries</option> <option value="behavsci">Behavioral Sciences</option> <option value="beverages">Beverages</option> <option value="BDCC">Big Data and Cognitive Computing</option> <option value="biochem">BioChem</option> <option value="bioengineering">Bioengineering</option> <option value="biologics">Biologics</option> <option value="biology">Biology</option> <option value="blsf">Biology and Life Sciences Forum</option> <option value="biomass">Biomass</option> <option value="biomechanics">Biomechanics</option> <option value="biomed">BioMed</option> <option value="biomedicines">Biomedicines</option> <option value="biomedinformatics">BioMedInformatics</option> <option value="biomimetics">Biomimetics</option> <option value="biomolecules">Biomolecules</option> <option value="biophysica">Biophysica</option> <option value="biosensors">Biosensors</option> <option value="biotech">BioTech</option> <option value="birds">Birds</option> <option value="blockchains">Blockchains</option> <option value="brainsci">Brain Sciences</option> <option value="buildings">Buildings</option> <option value="businesses">Businesses</option> <option value="carbon">C</option> <option value="cancers">Cancers</option> <option value="cardiogenetics">Cardiogenetics</option> <option value="catalysts">Catalysts</option> <option value="cells">Cells</option> <option value="ceramics">Ceramics</option> <option value="challenges">Challenges</option> <option value="ChemEngineering">ChemEngineering</option> <option value="chemistry">Chemistry</option> <option value="chemproc">Chemistry Proceedings</option> <option value="chemosensors">Chemosensors</option> <option value="children">Children</option> <option value="chips">Chips</option> <option value="civileng">CivilEng</option> <option value="cleantechnol">Clean Technologies</option> <option value="climate">Climate</option> <option value="ctn">Clinical and Translational Neuroscience</option> <option value="clinbioenerg">Clinical Bioenergetics</option> <option value="clinpract">Clinics and Practice</option> <option value="clockssleep">Clocks & Sleep</option> <option value="coasts">Coasts</option> <option value="coatings">Coatings</option> <option value="colloids">Colloids and Interfaces</option> <option value="colorants">Colorants</option> <option value="commodities">Commodities</option> <option value="complications">Complications</option> <option value="compounds">Compounds</option> <option value="computation">Computation</option> <option value="csmf">Computer Sciences & Mathematics Forum</option> <option value="computers">Computers</option> <option value="condensedmatter">Condensed Matter</option> <option value="conservation">Conservation</option> <option value="constrmater">Construction Materials</option> <option value="cmd">Corrosion and Materials Degradation</option> <option value="cosmetics">Cosmetics</option> <option value="covid">COVID</option> <option value="crops">Crops</option> <option value="cryo">Cryo</option> <option value="cryptography">Cryptography</option> <option value="crystals">Crystals</option> <option value="cimb">Current Issues in Molecular Biology</option> <option value="curroncol">Current Oncology</option> <option value="dairy">Dairy</option> <option value="data">Data</option> <option value="dentistry">Dentistry Journal</option> <option value="dermato">Dermato</option> <option value="dermatopathology">Dermatopathology</option> <option value="designs">Designs</option> <option value="diabetology">Diabetology</option> <option value="diagnostics">Diagnostics</option> <option value="dietetics">Dietetics</option> <option value="digital">Digital</option> <option value="disabilities">Disabilities</option> <option value="diseases">Diseases</option> <option value="diversity">Diversity</option> <option value="dna">DNA</option> <option value="drones">Drones</option> <option value="ddc">Drugs and Drug Candidates</option> <option value="dynamics">Dynamics</option> <option value="earth">Earth</option> <option value="ecologies">Ecologies</option> <option value="econometrics">Econometrics</option> <option value="economies">Economies</option> <option value="education">Education Sciences</option> <option value="electricity">Electricity</option> <option value="electrochem">Electrochem</option> <option value="electronicmat">Electronic Materials</option> <option value="electronics">Electronics</option> <option value="ecm">Emergency Care and Medicine</option> <option value="encyclopedia">Encyclopedia</option> <option value="endocrines">Endocrines</option> <option value="energies">Energies</option> <option value="esa">Energy Storage and Applications</option> <option value="eng">Eng</option> <option value="engproc">Engineering Proceedings</option> <option value="entropy">Entropy</option> <option value="environsciproc">Environmental Sciences Proceedings</option> <option value="environments">Environments</option> <option value="epidemiologia">Epidemiologia</option> <option value="epigenomes">Epigenomes</option> <option value="ebj">European Burn Journal</option> <option value="ejihpe">European Journal of Investigation in Health, Psychology and Education</option> <option value="fermentation">Fermentation</option> <option value="fibers">Fibers</option> <option value="fintech">FinTech</option> <option value="fire">Fire</option> <option value="fishes">Fishes</option> <option value="fluids">Fluids</option> <option value="foods">Foods</option> <option value="forecasting">Forecasting</option> <option value="forensicsci">Forensic Sciences</option> <option value="forests">Forests</option> <option value="fossstud">Fossil Studies</option> <option value="foundations">Foundations</option> <option value="fractalfract">Fractal and Fractional</option> <option value="fuels">Fuels</option> <option value="future">Future</option> <option value="futureinternet">Future Internet</option> <option value="futurepharmacol">Future Pharmacology</option> <option value="futuretransp">Future Transportation</option> <option value="galaxies">Galaxies</option> <option value="games">Games</option> <option value="gases">Gases</option> <option value="gastroent">Gastroenterology Insights</option> <option value="gastrointestdisord">Gastrointestinal Disorders</option> <option value="gastronomy">Gastronomy</option> <option value="gels">Gels</option> <option value="genealogy">Genealogy</option> <option value="genes">Genes</option> <option value="geographies">Geographies</option> <option value="geohazards">GeoHazards</option> <option value="geomatics">Geomatics</option> <option value="geometry">Geometry</option> <option value="geosciences">Geosciences</option> <option value="geotechnics">Geotechnics</option> <option value="geriatrics">Geriatrics</option> <option value="glacies">Glacies</option> <option value="gucdd">Gout, Urate, and Crystal Deposition Disease</option> <option value="grasses">Grasses</option> <option value="hardware">Hardware</option> <option value="healthcare">Healthcare</option> <option value="hearts">Hearts</option> <option value="hemato">Hemato</option> <option value="hematolrep">Hematology Reports</option> <option value="heritage">Heritage</option> <option value="histories">Histories</option> <option value="horticulturae">Horticulturae</option> <option value="hospitals">Hospitals</option> <option value="humanities">Humanities</option> <option value="humans">Humans</option> <option value="hydrobiology">Hydrobiology</option> <option value="hydrogen">Hydrogen</option> <option value="hydrology">Hydrology</option> <option value="hygiene">Hygiene</option> <option value="immuno">Immuno</option> <option value="idr">Infectious Disease Reports</option> <option value="informatics">Informatics</option> <option value="information">Information</option> <option value="infrastructures">Infrastructures</option> <option value="inorganics">Inorganics</option> <option value="insects">Insects</option> <option value="instruments">Instruments</option> <option value="iic">Intelligent Infrastructure and Construction</option> <option value="ijerph">International Journal of Environmental Research and Public Health</option> <option value="ijfs">International Journal of Financial Studies</option> <option value="ijms">International Journal of Molecular Sciences</option> <option value="IJNS">International Journal of Neonatal Screening</option> <option value="ijpb">International Journal of Plant Biology</option> <option value="ijt">International Journal of Topology</option> <option value="ijtm">International Journal of Translational Medicine</option> <option value="ijtpp">International Journal of Turbomachinery, Propulsion and Power</option> <option value="ime">International Medical Education</option> <option value="inventions">Inventions</option> <option value="IoT">IoT</option> <option value="ijgi">ISPRS International Journal of Geo-Information</option> <option value="J">J</option> <option value="jal">Journal of Ageing and Longevity</option> <option value="jcdd">Journal of Cardiovascular Development and Disease</option> <option value="jcto">Journal of Clinical & Translational Ophthalmology</option> <option value="jcm">Journal of Clinical Medicine</option> <option value="jcs">Journal of Composites Science</option> <option value="jcp">Journal of Cybersecurity and Privacy</option> <option value="jdad">Journal of Dementia and Alzheimer's Disease</option> <option value="jdb">Journal of Developmental Biology</option> <option value="jeta">Journal of Experimental and Theoretical Analyses</option> <option value="jfb">Journal of Functional Biomaterials</option> <option value="jfmk">Journal of Functional Morphology and Kinesiology</option> <option value="jof">Journal of Fungi</option> <option value="jimaging">Journal of Imaging</option> <option value="jintelligence">Journal of Intelligence</option> <option value="jlpea">Journal of Low Power Electronics and Applications</option> <option value="jmmp">Journal of Manufacturing and Materials Processing</option> <option value="jmse">Journal of Marine Science and Engineering</option> <option value="jmahp">Journal of Market Access & Health Policy</option> <option value="jmp">Journal of Molecular Pathology</option> <option value="jnt">Journal of Nanotheranostics</option> <option value="jne">Journal of Nuclear Engineering</option> <option value="ohbm">Journal of Otorhinolaryngology, Hearing and Balance Medicine</option> <option value="jop">Journal of Parks</option> <option value="jpm">Journal of Personalized Medicine</option> <option value="jpbi">Journal of Pharmaceutical and BioTech Industry</option> <option value="jor">Journal of Respiration</option> <option value="jrfm">Journal of Risk and Financial Management</option> <option value="jsan">Journal of Sensor and Actuator Networks</option> <option value="joma">Journal of the Oman Medical Association</option> <option value="jtaer">Journal of Theoretical and Applied Electronic Commerce Research</option> <option value="jvd">Journal of Vascular Diseases</option> <option value="jox">Journal of Xenobiotics</option> <option value="jzbg">Journal of Zoological and Botanical Gardens</option> <option value="journalmedia">Journalism and Media</option> <option value="kidneydial">Kidney and Dialysis</option> <option value="kinasesphosphatases">Kinases and Phosphatases</option> <option value="knowledge">Knowledge</option> <option value="labmed">LabMed</option> <option value="laboratories">Laboratories</option> <option value="land">Land</option> <option value="languages">Languages</option> <option value="laws">Laws</option> <option value="life">Life</option> <option value="limnolrev">Limnological Review</option> <option value="lipidology">Lipidology</option> <option value="liquids">Liquids</option> <option value="literature">Literature</option> <option value="livers">Livers</option> <option value="logics">Logics</option> <option value="logistics">Logistics</option> <option value="lubricants">Lubricants</option> <option value="lymphatics">Lymphatics</option> <option value="make">Machine Learning and Knowledge Extraction</option> <option value="machines">Machines</option> <option value="macromol">Macromol</option> <option value="magnetism">Magnetism</option> <option value="magnetochemistry">Magnetochemistry</option> <option value="marinedrugs">Marine Drugs</option> <option value="materials">Materials</option> <option value="materproc">Materials Proceedings</option> <option value="mca">Mathematical and Computational Applications</option> <option value="mathematics">Mathematics</option> <option value="medsci">Medical Sciences</option> <option value="msf">Medical Sciences Forum</option> <option value="medicina">Medicina</option> <option value="medicines">Medicines</option> <option value="membranes">Membranes</option> <option value="merits">Merits</option> <option value="metabolites">Metabolites</option> <option value="metals">Metals</option> <option value="meteorology">Meteorology</option> <option value="methane">Methane</option> <option value="mps">Methods and Protocols</option> <option value="metrics">Metrics</option> <option value="metrology">Metrology</option> <option value="micro">Micro</option> <option value="microbiolres">Microbiology Research</option> <option value="micromachines">Micromachines</option> <option value="microorganisms">Microorganisms</option> <option value="microplastics">Microplastics</option> <option value="minerals">Minerals</option> <option value="mining">Mining</option> <option value="modelling">Modelling</option> <option value="mmphys">Modern Mathematical Physics</option> <option value="molbank">Molbank</option> <option value="molecules">Molecules</option> <option value="mti">Multimodal Technologies and Interaction</option> <option value="muscles">Muscles</option> <option value="nanoenergyadv">Nanoenergy Advances</option> <option value="nanomanufacturing">Nanomanufacturing</option> <option value="nanomaterials">Nanomaterials</option> <option value="ndt">NDT</option> <option value="network">Network</option> <option value="neuroglia">Neuroglia</option> <option value="neurolint">Neurology International</option> <option value="neurosci">NeuroSci</option> <option value="nitrogen">Nitrogen</option> <option value="ncrna">Non-Coding RNA</option> <option value="nursrep">Nursing Reports</option> <option value="nutraceuticals">Nutraceuticals</option> <option value="nutrients">Nutrients</option> <option value="obesities">Obesities</option> <option value="oceans">Oceans</option> <option value="onco">Onco</option> <option value="optics">Optics</option> <option value="oral">Oral</option> <option value="organics">Organics</option> <option value="organoids">Organoids</option> <option value="osteology">Osteology</option> <option value="oxygen">Oxygen</option> <option value="parasitologia">Parasitologia</option> <option value="particles">Particles</option> <option value="pathogens">Pathogens</option> <option value="pathophysiology">Pathophysiology</option> <option value="pediatrrep">Pediatric Reports</option> <option value="pets">Pets</option> <option value="pharmaceuticals">Pharmaceuticals</option> <option value="pharmaceutics">Pharmaceutics</option> <option value="pharmacoepidemiology">Pharmacoepidemiology</option> <option value="pharmacy">Pharmacy</option> <option value="philosophies">Philosophies</option> <option value="photochem">Photochem</option> <option value="photonics">Photonics</option> <option value="phycology">Phycology</option> <option value="physchem">Physchem</option> <option value="psf">Physical Sciences Forum</option> <option value="physics">Physics</option> <option value="physiologia">Physiologia</option> <option value="plants">Plants</option> <option value="plasma">Plasma</option> <option value="platforms">Platforms</option> <option value="pollutants">Pollutants</option> <option value="polymers">Polymers</option> <option value="polysaccharides">Polysaccharides</option> <option value="populations">Populations</option> <option value="poultry">Poultry</option> <option value="powders">Powders</option> <option value="proceedings">Proceedings</option> <option value="processes">Processes</option> <option value="prosthesis">Prosthesis</option> <option value="proteomes">Proteomes</option> <option value="psychiatryint">Psychiatry International</option> <option value="psychoactives">Psychoactives</option> <option value="psycholint">Psychology International</option> <option value="publications">Publications</option> <option value="qubs">Quantum Beam Science</option> <option value="quantumrep">Quantum Reports</option> <option value="quaternary">Quaternary</option> <option value="radiation">Radiation</option> <option value="reactions">Reactions</option> <option value="realestate">Real Estate</option> <option value="receptors">Receptors</option> <option value="recycling">Recycling</option> <option value="rsee">Regional Science and Environmental Economics</option> <option value="religions">Religions</option> <option value="remotesensing">Remote Sensing</option> <option value="reports">Reports</option> <option value="reprodmed">Reproductive Medicine</option> <option value="resources">Resources</option> <option value="rheumato">Rheumato</option> <option value="risks">Risks</option> <option value="robotics">Robotics</option> <option value="ruminants">Ruminants</option> <option value="safety">Safety</option> <option value="sci">Sci</option> <option value="scipharm">Scientia Pharmaceutica</option> <option value="sclerosis">Sclerosis</option> <option value="seeds">Seeds</option> <option value="sensors">Sensors</option> <option value="separations">Separations</option> <option value="sexes">Sexes</option> <option value="signals">Signals</option> <option value="sinusitis">Sinusitis</option> <option value="smartcities">Smart Cities</option> <option value="socsci">Social Sciences</option> <option value="siuj">Société Internationale d’Urologie Journal</option> <option value="societies">Societies</option> <option value="software">Software</option> <option value="soilsystems">Soil Systems</option> <option value="solar">Solar</option> <option value="solids">Solids</option> <option value="spectroscj">Spectroscopy Journal</option> <option value="sports">Sports</option> <option value="standards">Standards</option> <option value="stats">Stats</option> <option value="stresses">Stresses</option> <option value="surfaces">Surfaces</option> <option value="surgeries">Surgeries</option> <option value="std">Surgical Techniques Development</option> <option value="sustainability">Sustainability</option> <option value="suschem">Sustainable Chemistry</option> <option value="symmetry">Symmetry</option> <option value="synbio">SynBio</option> <option value="systems">Systems</option> <option value="targets">Targets</option> <option value="taxonomy">Taxonomy</option> <option value="technologies">Technologies</option> <option value="telecom">Telecom</option> <option value="textiles">Textiles</option> <option value="thalassrep">Thalassemia Reports</option> <option value="therapeutics">Therapeutics</option> <option value="thermo">Thermo</option> <option value="timespace">Time and Space</option> <option value="tomography">Tomography</option> <option value="tourismhosp">Tourism and Hospitality</option> <option value="toxics">Toxics</option> <option value="toxins">Toxins</option> <option value="transplantology">Transplantology</option> <option value="traumacare">Trauma Care</option> <option value="higheredu">Trends in Higher Education</option> <option value="tropicalmed">Tropical Medicine and Infectious Disease</option> <option value="universe">Universe</option> <option value="urbansci">Urban Science</option> <option value="uro">Uro</option> <option value="vaccines">Vaccines</option> <option value="vehicles">Vehicles</option> <option value="venereology">Venereology</option> <option value="vetsci">Veterinary Sciences</option> <option value="vibration">Vibration</option> <option value="virtualworlds">Virtual Worlds</option> <option value="viruses">Viruses</option> <option value="vision">Vision</option> <option value="waste">Waste</option> <option value="water">Water</option> <option value="wild">Wild</option> <option value="wind">Wind</option> <option value="women">Women</option> <option value="world">World</option> <option value="wevj">World Electric Vehicle Journal</option> <option value="youth">Youth</option> <option value="zoonoticdis">Zoonotic Diseases</option> </select> <input name="email" type="email" placeholder="Enter your email address..." required="required" /> <button class="genericCaptcha button button--dark UA_FooterNewsletterSubscribeButton" type="submit">Subscribe</button> </form> </div> </div> </div> <div id="footer-copyright"> <div class="row"> <div class="columns large-6 medium-6 small-12 text-left"> © 1996-2024 MDPI (Basel, Switzerland) unless otherwise stated </div> <div class="columns large-6 medium-6 small-12 small-text-left medium-text-right large-text-right"> <a data-dropdown="drop-view-disclaimer" aria-controls="drop-view-disclaimer" aria-expanded="false" data-options="align:top; is_hover:true; hover_timeout:2000;"> Disclaimer </a> <div id="drop-view-disclaimer" class="f-dropdown label__btn__dropdown label__btn__dropdown--wide text-left" data-dropdown-content aria-hidden="true" tabindex="-1"> Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. </div> <a href="/about/terms-and-conditions"> Terms and Conditions </a> <a href="/about/privacy"> Privacy Policy </a> </div> </div> </div> </div> <div id="cookie-notification" class="js-allow-cookies" style="display: none;"> <div class="columns large-10 medium-10 small-12"> We use cookies on our website to ensure you get the best experience.<br class="show-for-medium-up"/> Read more about our cookies <a href="/about/privacy">here</a>. </div> <div class="columns large-2 medium-2 small-12 small-only-text-left text-right"> <a class="button button--default" href="/accept_cookies">Accept</a> </div> </div> </div> <div id="main-share-modal" class="reveal-modal reveal-modal-new reveal-modal-new--small" data-reveal aria-labelledby="modalTitle" aria-hidden="true" role="dialog"> <div class="row"> <div class="small-12 columns"> <h2 style="margin: 0;">Share Link</h2> </div> <div class="small-12 columns"> <div class="social-media-links UA_ShareModalLinks" style="text-align: left;"> <a href="/cdn-cgi/l/email-protection#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" title="Email"> <i class="fa fa-envelope-square" style="font-size: 30px;"></i> </a> <a href="https://twitter.com/intent/tweet?text=Kalata+B1+Enhances+Temozolomide+Toxicity+to+Glioblastoma+Cells&hashtags=mdpibiomedicines&url=https%3A%2F%2Fwww.mdpi.com%2F2976274&via=Biomed_MDPI" onclick="windowOpen(this.href,600,800); return false" title="Twitter" target="_blank" rel="noopener noreferrer"> <i class="fa fa-twitter-x-square" style="font-size: 30px;"></i> </a> <a href=" http://www.linkedin.com/shareArticle?mini=true&url=https%3A%2F%2Fwww.mdpi.com%2F2976274&title=Kalata%20B1%20Enhances%20Temozolomide%20Toxicity%20to%20Glioblastoma%20Cells%26source%3Dhttps%3A%2F%2Fwww.mdpi.com%26summary%3DGlioblastoma%20%28GBM%29%20is%20the%20most%20aggressive%20cancer%20originating%20in%20the%20brain%2C%20but%20unfortunately%20combination%20treatments%20with%20resection%2C%20radiation%2C%20and%20chemotherapy%20are%20relatively%20ineffective.%20Therefore%2C%20novel%20methods%20of%20adjuvant%20therapy%20are%20critically%20%5B...%5D" onclick="windowOpen(this.href,600,800); return false" title="LinkedIn" target="_blank" rel="noopener noreferrer"> <i class="fa fa-linkedin-square" style="font-size: 30px;"></i> </a> <a href="https://www.facebook.com/sharer.php?u=https://www.mdpi.com/2976274" title="facebook" target="_blank" rel="noopener noreferrer"> <i class="fa fa-facebook-square" style="font-size: 30px;"></i> </a> <a href="javascript:void(0);" title="Wechat" data-reveal-id="weixin-share-modal"> <i class="fa fa-weixin-square" style="font-size: 26px;"></i> </a> <a href="http://www.reddit.com/submit?url=https://www.mdpi.com/2976274" title="Reddit" target="_blank" rel="noopener noreferrer"> <i class="fa fa-reddit-square" style="font-size: 30px;"></i> </a> <a href="http://www.mendeley.com/import/?url=https://www.mdpi.com/2976274" title="Mendeley" target="_blank" rel="noopener noreferrer"> <i class="fa fa-mendeley-square" style="font-size: 30px;"></i> </a> <a href="http://www.citeulike.org/posturl?url=https://www.mdpi.com/2976274" title="CiteULike" target="_blank" rel="noopener noreferrer"> <i class="fa fa-citeulike-square" style="font-size: 30px;"></i> </a> </div> </div> <div class="small-9 columns"> <input id="js-clipboard-text" type="text" readonly value="https://www.mdpi.com/2976274" /> </div> <div class="small-3 columns text-left"> <a class="button button--color js-clipboard-copy" data-clipboard-target="#js-clipboard-text">Copy</a> </div> </div> <a class="close-reveal-modal" aria-label="Close"> <i class="material-icons">clear</i> </a> </div> <div id="weixin-share-modal" class="reveal-modal reveal-modal-new" data-reveal aria-labelledby="weixin-share-modal-title" aria-hidden="true" role="dialog"> <div class="row"> <div class="small-12 columns"> <h2 id="weixin-share-modal-title" style="margin: 0;">Share</h2> </div> <div class="small-12 columns"> <div class="weixin-qr-code-section"> <?xml version="1.0" standalone="no"?> <!DOCTYPE svg PUBLIC "-//W3C//DTD SVG 1.1//EN" "http://www.w3.org/Graphics/SVG/1.1/DTD/svg11.dtd"> <svg width="300" height="300" version="1.1" xmlns="http://www.w3.org/2000/svg"> <desc>https://www.mdpi.com/2976274</desc> <g id="elements" fill="black" stroke="none"> <rect x="0" y="0" width="12" height="12" /> <rect x="12" y="0" width="12" height="12" /> <rect x="24" y="0" width="12" height="12" /> <rect x="36" y="0" width="12" height="12" /> <rect x="48" y="0" width="12" height="12" /> <rect x="60" y="0" width="12" height="12" /> <rect x="72" y="0" width="12" height="12" /> <rect x="120" y="0" width="12" height="12" /> <rect x="168" y="0" width="12" height="12" /> <rect x="180" y="0" width="12" height="12" /> <rect x="216" y="0" width="12" height="12" /> <rect x="228" y="0" width="12" height="12" /> <rect x="240" y="0" width="12" height="12" /> <rect x="252" y="0" width="12" height="12" /> <rect x="264" y="0" width="12" height="12" /> <rect x="276" y="0" width="12" height="12" /> <rect x="288" y="0" width="12" height="12" /> <rect x="0" y="12" width="12" height="12" /> <rect x="72" y="12" width="12" height="12" /> <rect x="108" y="12" width="12" height="12" /> <rect x="180" y="12" width="12" height="12" /> <rect x="216" y="12" width="12" height="12" /> <rect x="288" y="12" width="12" height="12" /> <rect x="0" y="24" width="12" height="12" /> <rect x="24" y="24" width="12" height="12" /> <rect x="36" y="24" width="12" height="12" /> <rect x="48" y="24" width="12" height="12" /> <rect x="72" y="24" width="12" height="12" /> <rect x="96" y="24" width="12" height="12" /> <rect x="132" y="24" width="12" height="12" /> <rect x="180" y="24" width="12" height="12" /> <rect x="216" y="24" width="12" height="12" /> <rect x="240" y="24" width="12" height="12" /> <rect x="252" y="24" width="12" height="12" /> <rect x="264" y="24" width="12" height="12" /> <rect x="288" y="24" width="12" height="12" /> <rect x="0" y="36" width="12" height="12" /> <rect x="24" y="36" width="12" height="12" /> <rect x="36" y="36" width="12" height="12" /> <rect x="48" y="36" width="12" height="12" /> <rect x="72" y="36" width="12" height="12" /> <rect x="108" y="36" width="12" height="12" /> <rect x="168" y="36" width="12" height="12" /> <rect x="216" y="36" width="12" height="12" /> <rect x="240" y="36" width="12" height="12" /> <rect x="252" y="36" width="12" height="12" /> <rect x="264" y="36" width="12" height="12" /> <rect x="288" y="36" width="12" height="12" /> <rect x="0" y="48" width="12" height="12" /> <rect x="24" y="48" width="12" height="12" /> <rect x="36" y="48" width="12" height="12" /> <rect x="48" y="48" width="12" height="12" /> <rect x="72" y="48" width="12" height="12" /> <rect x="108" y="48" width="12" height="12" /> <rect x="144" y="48" width="12" height="12" /> <rect x="156" y="48" width="12" height="12" /> <rect x="168" y="48" width="12" height="12" /> <rect x="180" y="48" width="12" height="12" /> <rect x="192" y="48" width="12" height="12" /> <rect x="216" y="48" width="12" height="12" /> <rect x="240" y="48" width="12" height="12" /> <rect x="252" y="48" width="12" height="12" /> <rect x="264" y="48" width="12" height="12" /> <rect x="288" y="48" width="12" height="12" /> <rect x="0" y="60" width="12" height="12" /> <rect x="72" y="60" width="12" height="12" /> <rect x="120" y="60" width="12" height="12" /> <rect x="132" y="60" width="12" height="12" /> <rect x="168" y="60" width="12" height="12" /> <rect x="180" y="60" width="12" height="12" /> <rect x="192" y="60" width="12" height="12" /> <rect x="216" y="60" width="12" height="12" /> <rect x="288" y="60" width="12" height="12" /> <rect x="0" y="72" width="12" height="12" /> <rect x="12" y="72" width="12" height="12" /> <rect x="24" y="72" width="12" height="12" /> <rect x="36" y="72" width="12" height="12" /> <rect x="48" y="72" width="12" height="12" /> <rect x="60" y="72" width="12" height="12" /> <rect x="72" y="72" width="12" height="12" /> <rect x="96" y="72" width="12" height="12" /> <rect x="120" y="72" width="12" height="12" /> <rect x="144" y="72" width="12" height="12" /> <rect x="168" y="72" width="12" height="12" /> <rect x="192" y="72" width="12" height="12" /> <rect x="216" y="72" width="12" height="12" /> <rect x="228" y="72" width="12" height="12" /> <rect x="240" y="72" width="12" height="12" /> <rect x="252" y="72" width="12" height="12" /> <rect x="264" y="72" width="12" height="12" /> <rect x="276" y="72" width="12" height="12" /> <rect x="288" y="72" width="12" height="12" /> <rect x="96" y="84" width="12" height="12" /> <rect x="132" y="84" width="12" height="12" /> <rect x="156" y="84" width="12" height="12" /> <rect x="168" y="84" width="12" height="12" /> <rect x="180" y="84" width="12" height="12" /> <rect x="0" y="96" width="12" height="12" /> <rect x="12" y="96" width="12" height="12" /> <rect x="24" y="96" width="12" height="12" /> <rect x="48" y="96" width="12" height="12" /> <rect x="60" y="96" width="12" height="12" /> <rect x="72" y="96" width="12" height="12" /> <rect x="84" y="96" width="12" height="12" /> <rect x="96" y="96" width="12" height="12" /> <rect x="144" y="96" width="12" height="12" /> <rect x="156" y="96" width="12" height="12" /> <rect x="180" y="96" width="12" height="12" /> <rect x="192" y="96" width="12" height="12" /> <rect x="204" y="96" width="12" height="12" /> <rect x="216" y="96" width="12" height="12" /> <rect x="264" y="96" width="12" height="12" /> <rect x="0" y="108" width="12" height="12" /> <rect x="108" y="108" width="12" height="12" /> <rect x="132" y="108" width="12" height="12" /> <rect x="192" y="108" width="12" height="12" /> <rect x="216" y="108" width="12" height="12" /> <rect x="288" y="108" width="12" height="12" /> <rect x="48" y="120" width="12" height="12" /> <rect x="72" y="120" width="12" height="12" /> <rect x="84" y="120" width="12" height="12" /> <rect x="120" y="120" width="12" height="12" /> <rect x="132" y="120" width="12" height="12" /> <rect x="192" y="120" width="12" height="12" /> <rect x="204" y="120" width="12" height="12" /> <rect x="216" y="120" width="12" height="12" /> <rect x="228" y="120" width="12" height="12" /> <rect x="264" y="120" width="12" height="12" /> <rect x="276" y="120" width="12" height="12" /> <rect x="288" y="120" width="12" height="12" /> <rect x="0" y="132" width="12" height="12" /> <rect x="12" y="132" width="12" height="12" /> <rect x="24" y="132" width="12" height="12" /> <rect x="36" y="132" width="12" height="12" /> <rect x="48" y="132" width="12" height="12" /> <rect x="108" y="132" width="12" height="12" /> <rect x="120" y="132" width="12" height="12" /> <rect x="144" y="132" width="12" height="12" /> <rect x="156" y="132" width="12" height="12" /> <rect x="168" y="132" width="12" height="12" /> <rect x="180" y="132" width="12" height="12" /> <rect x="216" y="132" width="12" height="12" /> <rect x="276" y="132" width="12" height="12" /> <rect x="24" y="144" width="12" height="12" /> <rect x="48" y="144" width="12" height="12" /> <rect x="72" y="144" width="12" height="12" /> <rect x="96" y="144" width="12" height="12" /> <rect x="120" y="144" width="12" height="12" /> <rect x="132" y="144" width="12" height="12" /> <rect x="156" y="144" width="12" height="12" /> <rect x="168" y="144" width="12" height="12" /> <rect x="204" y="144" width="12" height="12" /> <rect x="216" y="144" width="12" height="12" /> <rect x="228" y="144" width="12" height="12" /> <rect x="252" y="144" width="12" height="12" /> <rect x="276" y="144" width="12" height="12" /> <rect x="288" y="144" width="12" height="12" /> <rect x="36" y="156" width="12" height="12" /> <rect x="84" y="156" width="12" height="12" /> <rect x="120" y="156" width="12" height="12" /> <rect x="132" y="156" width="12" height="12" /> <rect x="156" y="156" width="12" height="12" /> <rect x="192" y="156" width="12" height="12" /> <rect x="216" y="156" width="12" height="12" /> <rect x="252" y="156" width="12" height="12" /> <rect x="288" y="156" width="12" height="12" /> <rect x="0" y="168" width="12" height="12" /> <rect x="36" y="168" width="12" height="12" /> <rect x="48" y="168" width="12" height="12" /> <rect x="72" y="168" width="12" height="12" /> <rect x="96" y="168" width="12" height="12" /> <rect x="108" y="168" width="12" height="12" /> <rect x="120" y="168" width="12" height="12" /> <rect x="132" y="168" width="12" height="12" /> <rect x="144" y="168" width="12" height="12" /> <rect x="168" y="168" width="12" height="12" /> <rect x="192" y="168" width="12" height="12" /> <rect x="204" y="168" width="12" height="12" /> <rect x="216" y="168" width="12" height="12" /> <rect x="228" y="168" width="12" height="12" /> <rect x="264" y="168" width="12" height="12" /> <rect x="276" y="168" width="12" height="12" /> <rect x="288" y="168" width="12" height="12" /> <rect x="12" y="180" width="12" height="12" /> <rect x="60" y="180" width="12" height="12" /> <rect x="84" y="180" width="12" height="12" /> <rect x="96" y="180" width="12" height="12" /> <rect x="108" y="180" width="12" height="12" /> <rect x="120" y="180" width="12" height="12" /> <rect x="132" y="180" width="12" height="12" /> <rect x="156" y="180" width="12" height="12" /> <rect x="180" y="180" width="12" height="12" /> <rect x="192" y="180" width="12" height="12" /> <rect x="216" y="180" width="12" height="12" /> <rect x="240" y="180" width="12" height="12" /> <rect x="276" y="180" width="12" height="12" /> <rect x="0" y="192" width="12" height="12" /> <rect x="24" y="192" width="12" height="12" /> <rect x="36" y="192" width="12" height="12" /> <rect x="48" y="192" width="12" height="12" /> <rect x="72" y="192" width="12" height="12" /> <rect x="84" y="192" width="12" height="12" /> <rect x="96" y="192" width="12" height="12" /> <rect x="108" y="192" width="12" height="12" /> <rect x="144" y="192" width="12" height="12" /> <rect x="192" y="192" width="12" height="12" /> <rect x="204" y="192" width="12" height="12" /> <rect x="216" y="192" width="12" height="12" /> <rect x="228" y="192" width="12" height="12" /> <rect x="240" y="192" width="12" height="12" /> <rect x="252" y="192" width="12" height="12" /> <rect x="96" y="204" width="12" height="12" /> <rect x="108" y="204" width="12" height="12" /> <rect x="120" y="204" width="12" height="12" /> <rect x="132" y="204" width="12" height="12" /> <rect x="144" y="204" width="12" height="12" /> <rect x="156" y="204" width="12" height="12" /> <rect x="180" y="204" width="12" height="12" /> <rect x="192" y="204" width="12" height="12" /> <rect x="240" y="204" width="12" height="12" /> <rect x="252" y="204" width="12" height="12" /> <rect x="276" y="204" width="12" height="12" /> <rect x="288" y="204" width="12" height="12" /> <rect x="0" y="216" width="12" height="12" /> <rect x="12" y="216" width="12" height="12" /> <rect x="24" y="216" width="12" height="12" /> <rect x="36" y="216" width="12" height="12" /> <rect x="48" y="216" width="12" height="12" /> <rect x="60" y="216" width="12" height="12" /> <rect x="72" y="216" width="12" height="12" /> <rect x="96" y="216" width="12" height="12" /> <rect x="108" y="216" width="12" height="12" /> <rect x="144" y="216" width="12" height="12" /> <rect x="156" y="216" width="12" height="12" /> <rect x="180" y="216" width="12" height="12" /> <rect x="192" y="216" width="12" height="12" /> <rect x="216" y="216" width="12" height="12" /> <rect x="240" y="216" width="12" height="12" /> <rect x="252" y="216" width="12" height="12" /> <rect x="276" y="216" width="12" height="12" /> <rect x="288" y="216" width="12" height="12" /> <rect x="0" y="228" width="12" height="12" /> <rect x="72" y="228" width="12" height="12" /> <rect x="96" y="228" width="12" height="12" /> <rect x="120" y="228" width="12" height="12" /> <rect x="132" y="228" width="12" height="12" /> <rect x="144" y="228" width="12" height="12" /> <rect x="168" y="228" width="12" height="12" /> <rect x="192" y="228" width="12" height="12" /> <rect x="240" y="228" width="12" height="12" /> <rect x="252" y="228" width="12" height="12" /> <rect x="276" y="228" width="12" height="12" /> <rect x="0" y="240" width="12" height="12" /> <rect x="24" y="240" width="12" height="12" /> <rect x="36" y="240" width="12" height="12" /> <rect x="48" y="240" width="12" height="12" /> <rect x="72" y="240" width="12" height="12" /> <rect x="96" y="240" width="12" height="12" /> <rect x="120" y="240" width="12" height="12" /> <rect x="144" y="240" width="12" height="12" /> <rect x="168" y="240" width="12" height="12" /> <rect x="192" y="240" width="12" height="12" /> <rect x="204" y="240" width="12" height="12" /> <rect x="216" y="240" width="12" height="12" /> <rect x="228" y="240" width="12" height="12" /> <rect x="240" y="240" width="12" height="12" /> <rect x="252" y="240" width="12" height="12" /> <rect x="276" y="240" width="12" height="12" /> <rect x="288" y="240" width="12" height="12" /> <rect x="0" y="252" width="12" height="12" /> <rect x="24" y="252" width="12" height="12" /> <rect x="36" y="252" width="12" height="12" /> <rect x="48" y="252" width="12" height="12" /> <rect x="72" y="252" width="12" height="12" /> <rect x="108" y="252" width="12" height="12" /> <rect x="120" y="252" width="12" height="12" /> <rect x="132" y="252" width="12" height="12" /> <rect x="156" y="252" width="12" height="12" /> <rect x="168" y="252" width="12" height="12" /> <rect x="204" y="252" width="12" height="12" /> <rect x="228" y="252" width="12" height="12" /> <rect x="240" y="252" width="12" height="12" /> <rect x="252" y="252" width="12" height="12" /> <rect x="264" y="252" width="12" height="12" /> <rect x="0" y="264" width="12" height="12" /> <rect x="24" y="264" width="12" height="12" /> <rect x="36" y="264" width="12" height="12" /> <rect x="48" y="264" width="12" height="12" /> <rect x="72" y="264" width="12" height="12" /> <rect x="96" y="264" width="12" height="12" /> <rect x="108" y="264" width="12" height="12" /> <rect x="132" y="264" width="12" height="12" /> <rect x="168" y="264" width="12" height="12" /> <rect x="180" y="264" width="12" height="12" /> <rect x="240" y="264" width="12" height="12" /> <rect x="288" y="264" width="12" height="12" /> <rect x="0" y="276" width="12" height="12" /> <rect x="72" y="276" width="12" height="12" /> <rect x="96" y="276" width="12" height="12" /> <rect x="108" y="276" width="12" height="12" /> <rect x="156" y="276" width="12" height="12" /> <rect x="168" y="276" width="12" height="12" /> <rect x="180" y="276" width="12" height="12" /> <rect x="192" y="276" width="12" height="12" /> <rect x="216" y="276" width="12" height="12" /> <rect x="240" y="276" width="12" height="12" /> <rect x="252" y="276" width="12" height="12" /> <rect x="276" y="276" width="12" height="12" /> <rect x="0" y="288" width="12" height="12" /> <rect x="12" y="288" width="12" height="12" /> <rect x="24" y="288" width="12" height="12" /> <rect x="36" y="288" width="12" height="12" /> <rect x="48" y="288" width="12" height="12" /> <rect x="60" y="288" width="12" height="12" /> <rect x="72" y="288" width="12" height="12" /> <rect x="96" y="288" width="12" height="12" /> <rect x="108" y="288" width="12" height="12" /> <rect x="156" y="288" width="12" height="12" /> <rect x="168" y="288" width="12" height="12" /> <rect x="180" y="288" width="12" height="12" /> <rect x="192" y="288" width="12" height="12" /> <rect x="228" y="288" width="12" height="12" /> <rect x="276" y="288" width="12" height="12" /> <rect x="288" y="288" width="12" height="12" /> </g> </svg> </div> </div> </div> <a class="close-reveal-modal" aria-label="Close"> <i class="material-icons">clear</i> </a> </div> <a href="#" class="back-to-top"><span class="show-for-medium-up">Back to Top</span><span class="show-for-small">Top</span></a> <script data-cfasync="false" src="/cdn-cgi/scripts/5c5dd728/cloudflare-static/email-decode.min.js"></script><script src="https://pub.mdpi-res.com/assets/js/modernizr-2.8.3.min.js?5227e0738f7f421d?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/jquery-1.12.4.min.js?4f252523d4af0b47?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/foundation-5.5.3.min.js?6b2ec41c18b29054?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/foundation-5.5.3.equalizer.min.js?0f6c549b75ec554c?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/jquery.multiselect.js?0edd3998731d1091?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/jquery.cycle2.min.js?63413052928f97ee?1732615622"></script> <script> // old browser fix - this way the console log rows won't throw (silent) errors in browsers not supporting console log if (!window.console) window.console = {}; if (!window.console.log) window.console.log = function () { }; var currentJournalNameSystem = "biomedicines"; $(document).ready(function() { $('select.foundation-select').multiselect({ search: true, minHeight: 130, maxHeight: 130, }); $(document).foundation({ orbit: { timer_speed: 4000, }, reveal: { animation: 'fadeAndPop', animation_speed: 100, } }); $(".chosen-select").each(function(element) { var maxSelected = (undefined !== $(this).data('maxselectedoptions') ? $(this).data('maxselectedoptions') : 100); $(this).on('chosen:ready', function(event, data) { var select = $(data.chosen.form_field); if (select.attr('id') === 'journal-browser-volume') { $(data.chosen.dropdown).addClass('UI_JournalBrowser_Volume_Options'); } if (select.attr('id') === 'journal-browser-issue') { $(data.chosen.dropdown).addClass('UI_JournalBrowser_Issue_Options'); } }).chosen({ display_disabled_options: false, disable_search_threshold: 7, max_selected_options: maxSelected, width: "100%" }); }); $(".toEncode").each(function(e) { var oldHref = $(this).attr("href"); var newHref = oldHref.replace('.botdefense.please.enable.javascript.','@'); $(this).attr("href", newHref); if (!$(this).hasClass("emailCaptcha")) { $(this).html(newHref.replace('mailto:', '')); } $(this).removeClass("visibility-hidden"); }); $(document).on('opened.fndtn.reveal', '[data-reveal]', function() { $(document).foundation('equalizer', 'reflow'); }); // fix the images that have tag height / width defined // otherwise the default foundation styles overwrite the tag definitions $("img").each(function() { if ($(this).attr('width') != undefined || $(this).attr('height') != undefined) { $(this).addClass("img-fixed"); } }); $("#basic_search, #advanced_search").submit(function(e) { var searchArguments = false; $(this).find("input,select").not("#search,.search-button").each(function() { if (undefined === $(this).val() || "" === $(this).val()) { $(this).attr('name', null); } else { $(this).attr('name'); searchArguments = true; } }); if (!searchArguments) { window.location = $(this).attr('action'); return false; } }); $(".hide-show-desktop-option").click(function(e) { e.preventDefault(); var parentDiv = $(this).closest("div"); $.ajax({ url: $(this).attr('href'), success: function(msg) { parentDiv.removeClass().hide(); } }); }); $(".generic-toggleable-header").click(function(e) { $(this).toggleClass("active"); $(this).next(".generic-toggleable-content").toggleClass("active"); }); /* * handle whole row as a link if the row contains only one visible link */ $("table.new tr").hover(function() { if ($(this).find("td:visible a").length == 1) { $(this).addClass("single-link"); } }, function() { $(this).removeClass("single-link"); }); $("table.new:not(.table-of-tables)").on("click", "tr.single-link", function(e) { var target = $(e.target); if (!e.ctrlKey && !target.is("a")) { $(this).find("td:visible a")[0].click(); } }); $(document).on("click", ".custom-accordion-for-small-screen-link", function(e) { if ($(this).closest("#basic_search").length > 0) { if ($(".search-container__advanced").first().is(":visible")) { openAdvanced() } } if (Foundation.utils.is_small_only()) { if ($(this).hasClass("active")) { $(this).removeClass("active"); $(this).next(".custom-accordion-for-small-screen-content").addClass("show-for-medium-up"); } else { $(this).addClass("active"); $(this).next(".custom-accordion-for-small-screen-content").removeClass("show-for-medium-up"); $(document).foundation('orbit', 'reflow'); } } if (undefined !== $(this).data("callback")) { var customCallback = $(this).data("callback"); func = window[customCallback]; func(); } }); $(document).on("click", ".js-open-small-search", function(e) { e.preventDefault(); $(this).toggleClass("active").closest(".tab-bar").toggleClass("active"); $(".search-container").toggleClass("hide-for-small-down"); }); $(document).on("click", ".js-open-menu", function(e) { $(".search-container").addClass("hide-for-small-down"); }); $(window).on('resize', function() { recalculate_main_browser_position(); recalculate_responsive_moving_containers(); }); updateSearchLabelVisibilities(); recalculate_main_browser_position(); recalculate_responsive_moving_containers(); if (window.document.documentMode == 11) { $("<link/>", { rel: "stylesheet", type: "text/css", href: "https://fonts.googleapis.com/icon?family=Material+Icons"}).appendTo("head"); } }); function recalculate_main_browser_position() { if (Foundation.utils.is_small_only()) { if ($("#js-main-top-container").parent("#js-large-main-top-container").length > 0) { $("#js-main-top-container").appendTo($("#js-small-main-top-container")); } } else { if ($("#js-main-top-container").parent("#js-small-main-top-container").length > 0) { $("#js-main-top-container").appendTo($("#js-large-main-top-container")); } } } function recalculate_responsive_moving_containers() { $(".responsive-moving-container.large").each(function() { var previousParent = $(".responsive-moving-container.active[data-id='"+$(this).data("id")+"']"); var movingContent = previousParent.html(); if (Foundation.utils.is_small_only()) { var currentParent = $(".responsive-moving-container.small[data-id='"+$(this).data("id")+"']"); } else if (Foundation.utils.is_medium_only()) { var currentParent = $(".responsive-moving-container.medium[data-id='"+$(this).data("id")+"']"); } else { var currentParent = $(".responsive-moving-container.large[data-id='"+$(this).data("id")+"']"); } if (previousParent.attr("class") !== currentParent.attr("class")) { currentParent.html(movingContent); previousParent.html(); currentParent.addClass("active"); previousParent.removeClass("active"); } }); } // cookies allowed is checked from a) local storage and b) from server separately so that the footer bar doesn't // get included in the custom page caches function checkCookiesAllowed() { var cookiesEnabled = localStorage.getItem("mdpi_cookies_enabled"); if (null === cookiesEnabled) { $.ajax({ url: "/ajax_cookie_value/mdpi_cookies_accepted", success: function(data) { if (data.value) { localStorage.setItem("mdpi_cookies_enabled", true); checkDisplaySurvey(); } else { $(".js-allow-cookies").show(); } } }); } else { checkDisplaySurvey(); } } function checkDisplaySurvey() { } window.addEventListener('CookiebotOnAccept', function (e) { var CookieDate = new Date; if (Cookiebot.consent.preferences) { CookieDate.setFullYear(CookieDate.getFullYear() + 1); document.cookie = "mdpi_layout_type_v2=mobile; path=/; expires=" + CookieDate.toUTCString() + ";"; $(".js-toggle-desktop-layout-link").css("display", "inline-block"); } }, false); window.addEventListener('CookiebotOnDecline', function (e) { if (!Cookiebot.consent.preferences) { $(".js-toggle-desktop-layout-link").hide(); if ("" === "desktop") { window.location = "/toggle_desktop_layout_cookie"; } } }, false); var hash = $(location).attr('hash'); if ("#share" === hash) { if (1 === $("#main-share-modal").length) { $('#main-share-modal').foundation('reveal', 'open'); } } </script> <script src="https://pub.mdpi-res.com/assets/js/lib.js?f8d3d71b3a772f9d?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/mdpi.js?c267ce58392b15da?1732615622"></script> <script>var banners_url = 'https://serve.mdpi.com';</script> <script type='text/javascript' src='https://pub.mdpi-res.com/assets/js/ifvisible.min.js?c621d19ecb761212?1732615622'></script> <script src="https://pub.mdpi-res.com/assets/js/xmltohtml/affix.js?ac4ea55275297c15?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/clipboard.min.js?3f3688138a1b9fc4?1732615622"></script> <script type="text/javascript"> $(document).ready(function() { var helpFunctions = $(".middle-column__help__fixed"); var leftColumnAffix = $(".left-column__fixed"); var middleColumn = $("#middle-column"); var clone = null; helpFunctions.affix({ offset: { top: function() { return middleColumn.offset().top - 8 - (Foundation.utils.is_medium_only() ? 30 : 0); }, bottom: function() { return $("#footer").innerHeight() + 74 + (Foundation.utils.is_medium_only() ? 0 : 0); } } }); if (leftColumnAffix.length > 0) { clone = leftColumnAffix.clone(); clone.addClass("left-column__fixed__affix"); clone.insertBefore(leftColumnAffix); clone.css('width', leftColumnAffix.outerWidth() + 50); clone.affix({ offset: { top: function() { return leftColumnAffix.offset().top - 30 - (Foundation.utils.is_medium_only() ? 50 : 0); }, bottom: function() { return $("#footer").innerHeight() + 92 + (Foundation.utils.is_medium_only() ? 0 : 0); } } }); } $(window).on("resize", function() { if (clone !== null) { clone.css('width', leftColumnAffix.outerWidth() + 50); } }); new ClipboardJS('.js-clipboard-copy'); }); </script> <script src="https://pub.mdpi-res.com/assets/js/jquery-ui-1.13.2.min.js?1e2047978946a1d2?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/slick.min.js?d5a61c749e44e471?1732615622"></script> <script> $(document).ready(function() { $(".link-article-menu").click(function(e) { e.preventDefault(); $(this).find('span').toggle(); $(this).next("div").toggleClass("active"); }); $(".js-similarity-related-articles").click(function(e) { e.preventDefault(); if ('' !== $('#recommended-articles-modal').attr('data-url')) { $('#recommended-articles-modal').foundation('reveal', 'open', $('#recommended-articles-modal').attr('data-url')); } }); $.ajax({ url: "/article/1488137/similarity-related/show-link", success: function(result) { if (result.show) { $('#recommended-articles-modal').attr('data-url', result.link); $('.js-article-similarity-container').show(); } } }); $(document).on('opened.fndtn.reveal', '[data-reveal]', function() { var modal = $(this); if (modal.attr('id') === "author-biographies-modal") { modal.find('.multiple-items').slick({ slidesToShow: 1, nextArrow: '<a class="slick-next" href="#"><i class="material-icons">chevron_right</i></a>', prevArrow: '<a class="slick-prev" href="#"><i class="material-icons">chevron_left</i></a>', slidesToScroll: 1, draggable: false, }); modal.find('.multiple-items').slick('refresh'); } }); }); </script> <script> $(document).ready(function() { $(document).on('keyup', function (e) { if (e.keyCode == 27) { var hElem = $(this).find(".annotator-adder"); if (hElem.length){ hElem.css({'visibility':'hidden'}); } else { document.querySelector("hypothesis-adder").shadowRoot.querySelector(".annotator-adder").style.visibility = "hidden"; } } }); }); </script> <script> window.hypothesisConfig = function () { return { sidebarAppUrl: 'https://commenting.mdpi.com/app.html', showHighlights: 'whenSidebarOpen' , openSidebar: false , assetRoot: 'https://commentingres.mdpi.com/hypothesis', services: [{ apiUrl: 'https://commenting.mdpi.com/api/', authority: 'mdpi', grantToken: '', doi: '10.3390/biomedicines12102216' }], }; }; </script> <script async id="hypothesis_frame"></script> <script type="text/javascript"> if (-1 !== window.location.href.indexOf("?src=")) { window.history.replaceState({}, '', `${location.pathname}`); } $(document).ready(function() { var scifeedCounter = 0; var search = window.location.search; var mathjaxReady = false; // late image file loading $("img[data-lsrc]").each(function() { $(this).attr("src", $(this).data("lsrc")); }); // late mathjax initialization var head = document.getElementsByTagName("head")[0]; var script = document.createElement("script"); script.type = "text/x-mathjax-config"; script[(window.opera ? "innerHTML" : "text")] = "MathJax.Hub.processSectionDelay = 0;\n" + "MathJax.Hub.Config({\n" + " \"menuSettings\": {\n" + " CHTMLpreview: false\n" + " },\n" + " \"CHTML-preview\":{\n" + " disabled: true\n" + " },\n" + " \"HTML-CSS\": {\n" + " scale: 90,\n" + " availableFonts: [],\n" + " preferredFont: null,\n" + " preferredFonts: null,\n" + " webFont:\"Gyre-Pagella\",\n" + " imageFont:'TeX',\n" + " undefinedFamily:\"'Arial Unicode MS',serif\",\n" + " linebreaks: { automatic: false }\n" + " },\n" + " \"TeX\": {\n" + " extensions: ['noErrors.js'],\n" + " noErrors: {\n" + " inlineDelimiters: [\"\",\"\"],\n" + " multiLine: true,\n" + " style: {\n" + " 'font-size': '90%',\n" + " 'text-align': 'left',\n" + " 'color': 'black',\n" + " 'padding': '1px 3px',\n" + " 'border': '1px solid'\n" + " }\n" + " }\n" + " }\n" + "});\n" + "MathJax.Hub.Register.StartupHook('End', function() {\n" + " refreshMathjaxWidths();\n" + " mathjaxReady = true;\n" + "});\n" + "MathJax.Hub.Startup.signal.Interest(function (message) {\n" + " if (message == 'End') {\n" + " var hypoLink = document.getElementById('hypothesis_frame');\n" + " if (null !== hypoLink) {\n" + " hypoLink.setAttribute('src', 'https://commenting.mdpi.com/embed.js');\n" + " }\n" + " }\n" + "});"; head.appendChild(script); script = document.createElement("script"); script.type = "text/javascript"; script.src = "https://pub.mdpi-res.com/bundles/mathjax/MathJax.js?config=TeX-AMS-MML_HTMLorMML"; head.appendChild(script); // article version checker if (0 === search.indexOf('?type=check_update&version=')) { $.ajax({ url: "/2227-9059/12/10/2216" + "/versioncheck" + search, success: function(result) { $(".js-check-update-container").html(result); } }); } $('#feed_option').click(function() { // tracker if ($('#scifeed_clicked').length<1) { $(this).append('<span style="display:none" id="scifeed_clicked">done</span>'); } $('#feed_data').toggle('slide', { direction: 'up'}, '1000'); // slideToggle(700); OR toggle(700) $("#scifeed_error_msg").html('').hide(); $("#scifeed_notice_msg").html('').hide(); }); $('#feed_option').click(function(event) { setTimeout(function(){ var captchaSection = $("#captchaSection"); captchaSection.removeClass('ui-helper-hidden').find('input').prop('disabled', false); // var img = captchaSection.find('img'); // img.attr('src', img.data('url') + "?" + (new Date()).getTime()); // $(".captcha_reload").trigger("click"); var img = document.getElementById('gregwar_captcha_scifeed'); img.src = '/generate-captcha/gcb_captcha?n=' + (new Date()).getTime(); },800); }); $(document).on('click', '.split_feeds', function() { var name = $( this ).attr('name'); var flag = 1 - ($(this).is(":checked")*1); $('.split_feeds').each(function (index) { if ($( this ).attr('name') !== name) { $(this)[0].checked = flag; } }); }); $(document).on('click', '#scifeed_submit, #scifeed_submit1', function(event) { event.preventDefault(); $(".captcha_reload").trigger("click"); $("#scifeed_error_msg").html(""); $("#scifeed_error_msg").hide(); }); $(document).on('click', '.subscription_toggle', function(event) { if ($(this).val() === 'Create SciFeed' && $('#scifeed_hidden_flag').length>0) { event.preventDefault(); // alert('Here there would be a captcha because user is not logged in'); var captchaSection = $("#captchaSection"); if (captchaSection.hasClass('ui-helper-hidden')) { captchaSection.removeClass('ui-helper-hidden').find('input').prop('disabled', false); var img = captchaSection.find('img'); img.attr('src', img.data('url') + "?" + (new Date()).getTime()); $("#reloadCaptcha").trigger("click"); } } }); $(document).on('click', '.scifeed_msg', function(){ $(this).hide(); }); $(document).on('click', '.article-scilit-search', function(e) { e.preventDefault(); var data = $(".article-scilit-search-data").val(); var dataArray = data.split(';').map(function(keyword) { return "(\"" + keyword.trim() + "\")"; }); var searchQuery = dataArray.join(" OR "); var searchUrl = encodeURI("https://www.scilit.net/articles/search?q="+ searchQuery + "&advanced=1&highlight=1"); var win = window.open(searchUrl, '_blank'); if (win) { win.focus(); } else { window.location(searchUrl); } }); display_stats(); citedCount(); follow_goto(); // Select the node that will be observed for mutations const targetNodes = document.getElementsByClassName('hypothesis-count-container'); // Options for the observer (which mutations to observe) const config = { attributes: false, childList: true, subtree: false }; // Callback function to execute when mutations are observed const callback = function(mutationList, observer) { for(const mutation of mutationList) { if (mutation.type === 'childList') { let node = $(mutation.target); if (parseInt(node.html()) > 0) { node.show(); } } } }; // Create an observer instance linked to the callback function const observer = new MutationObserver(callback); // Start observing the target node for configured mutations for(const targetNode of targetNodes) { observer.observe(targetNode, config); } // Select the node that will be observed for mutations const mathjaxTargetNode = document.getElementById('middle-column'); // Callback function to execute when mutations are observed const mathjaxCallback = function(mutationList, observer) { if (mathjaxReady && typeof(MathJax) !== 'undefined') { refreshMathjaxWidths(); } }; // Create an observer instance linked to the callback function const mathjaxObserver = new ResizeObserver(mathjaxCallback); // Start observing the target node for configured mutations mathjaxObserver.observe(mathjaxTargetNode); }); /* END $(document).ready */ function refreshMathjaxWidths() { let width = ($('.html-body').width()*0.9) + "px"; $('.MathJax_Display').css('max-width', width); $('.MJXc-display').css('max-width', width); } function sendScifeedFrom(form) { if (!$('#scifeed_email').val().trim()) { // empty email alert('Please, provide an email for subscribe to this scifeed'); return false; } else if (!$('#captchaSection').hasClass('ui-helper-hidden') && !$('#captchaSection').find('input').val().trim()) { // empty captcha alert('Please, fill the captcha field.'); return false; } else if( ((($('#scifeed_form').find('input:checkbox:checked').length)-($('#split_feeds:checked').length))<1) || ($('#scifeed_kwd_txt').length < 0 && !$('#scifeed_kwd_txt').val().trim()) || ($('#scifeed_author_txt').length<0 &&!$('#scifeed_author_txt').val().trim()) ) { alert('You did not select anything to subscribe'); return false; } else if(($('#scifeed_form').find('input:checkbox:checked').length)-($('#split_feeds2:checked').length)<1){ alert("You did not select anything to subscribe"); return false; } else { var url = $('#scifeed_subscribe_url').html(); var formData = $(form).serializeArray(); $.post(url, formData).done(function (data) { if (JSON.parse(data)) { $('.scifeed_msg').hide(); var res = JSON.parse(data); var successFeeds = 0; var errorFeeds = 0; if (res) { $('.scifeed_msg').html(''); $.each(res, function (index, val) { if (val) { if (val.error) { errorFeeds++; $("#scifeed_error_msg").append(index+' - '+val.error+'<br>'); } if (val.notice) // for successful feed creation { successFeeds++; // $("#scifeed_notice_msg").append(index+' - '+val.notice+'<br>'); $("#scifeed_notice_msg").append('<li>'+index+'</li>'); } } }); if (successFeeds>0) { text = $('#scifeed_notice_msg').html(); text = 'The following feed'+(successFeeds>1?'s have':' has')+ ' been sucessfully created:<br><ul>'+ text + '</ul>' +($('#scifeed_hidden_flag').length>0 ? 'You are not logged in, so you probably need to validate '+ (successFeeds>1?'them':' it')+'.<br>' :'' ) +'Please check your email'+(successFeeds>1?'s':'')+' for more details.'; //(successFeeds>1?' for each of them':'')+'.<br>'; $("#scifeed_notice_msg").html(text); $("#scifeed_notice_msg").show(); } if (errorFeeds>0) { $("#scifeed_error_msg").show();; } } $("#feed_data").hide(); } }); } } function follow_goto() { var hashStr = location.hash.replace("#",""); if(typeof hashStr !== 'undefined') { if( hashStr == 'supplementary') { document.getElementById('suppl_id').scrollIntoView(); } if( hashStr == 'citedby') { document.getElementById('cited_id').scrollIntoView(); } } } function cited() { $("#framed_div").toggle('fast', function(){ if ($(this).css('display') != 'none') { var loaded = document.getElementById("loaded"); if(loaded.innerHTML == "No") { // Load Xref result var container = document.getElementById("framed_div"); // This replace the content container.innerHTML = "<img src=\"https://pub.mdpi-res.com/img/loading_circle.gif?9a82694213036313?1732615622\" height=\"20\" width=\"20\" alt=\"Processing...\" style=\"vertical-align:middle; margin-right:0.6em;\">"; var url = "/citedby/10.3390%252Fbiomedicines12102216/158"; $.post(url, function(result) { if (result.success) { container.innerHTML = result.view; } loaded.innerHTML = "Yes"; }); } } return true; // for not going at the beginning of the page... }) return true; // for not going at the beginning of the page... } function detect_device() { // Added by Bastien (18/08/2014): based on the http://detectmobilebrowsers.com/ detector var check = false; (function(a){if(/(android|bb\d+|meego).+mobile|avantgo|bada\/|blackberry|blazer|compal|elaine|fennec|hiptop|iemobile|ip(hone|od)|iris|kindle|lge |maemo|midp|mmp|mobile.+firefox|netfront|opera m(ob|in)i|palm( os)?|phone|p(ixi|re)\/|plucker|pocket|psp|series(4|6)0|symbian|treo|up\.(browser|link)|vodafone|wap|windows (ce|phone)|xda|xiino/i.test(a)||/1207|6310|6590|3gso|4thp|50[1-6]i|770s|802s|a wa|abac|ac(er|oo|s\-)|ai(ko|rn)|al(av|ca|co)|amoi|an(ex|ny|yw)|aptu|ar(ch|go)|as(te|us)|attw|au(di|\-m|r |s )|avan|be(ck|ll|nq)|bi(lb|rd)|bl(ac|az)|br(e|v)w|bumb|bw\-(n|u)|c55\/|capi|ccwa|cdm\-|cell|chtm|cldc|cmd\-|co(mp|nd)|craw|da(it|ll|ng)|dbte|dc\-s|devi|dica|dmob|do(c|p)o|ds(12|\-d)|el(49|ai)|em(l2|ul)|er(ic|k0)|esl8|ez([4-7]0|os|wa|ze)|fetc|fly(\-|_)|g1 u|g560|gene|gf\-5|g\-mo|go(\.w|od)|gr(ad|un)|haie|hcit|hd\-(m|p|t)|hei\-|hi(pt|ta)|hp( i|ip)|hs\-c|ht(c(\-| |_|a|g|p|s|t)|tp)|hu(aw|tc)|i\-(20|go|ma)|i230|iac( |\-|\/)|ibro|idea|ig01|ikom|im1k|inno|ipaq|iris|ja(t|v)a|jbro|jemu|jigs|kddi|keji|kgt( |\/)|klon|kpt |kwc\-|kyo(c|k)|le(no|xi)|lg( g|\/(k|l|u)|50|54|\-[a-w])|libw|lynx|m1\-w|m3ga|m50\/|ma(te|ui|xo)|mc(01|21|ca)|m\-cr|me(rc|ri)|mi(o8|oa|ts)|mmef|mo(01|02|bi|de|do|t(\-| |o|v)|zz)|mt(50|p1|v )|mwbp|mywa|n10[0-2]|n20[2-3]|n30(0|2)|n50(0|2|5)|n7(0(0|1)|10)|ne((c|m)\-|on|tf|wf|wg|wt)|nok(6|i)|nzph|o2im|op(ti|wv)|oran|owg1|p800|pan(a|d|t)|pdxg|pg(13|\-([1-8]|c))|phil|pire|pl(ay|uc)|pn\-2|po(ck|rt|se)|prox|psio|pt\-g|qa\-a|qc(07|12|21|32|60|\-[2-7]|i\-)|qtek|r380|r600|raks|rim9|ro(ve|zo)|s55\/|sa(ge|ma|mm|ms|ny|va)|sc(01|h\-|oo|p\-)|sdk\/|se(c(\-|0|1)|47|mc|nd|ri)|sgh\-|shar|sie(\-|m)|sk\-0|sl(45|id)|sm(al|ar|b3|it|t5)|so(ft|ny)|sp(01|h\-|v\-|v )|sy(01|mb)|t2(18|50)|t6(00|10|18)|ta(gt|lk)|tcl\-|tdg\-|tel(i|m)|tim\-|t\-mo|to(pl|sh)|ts(70|m\-|m3|m5)|tx\-9|up(\.b|g1|si)|utst|v400|v750|veri|vi(rg|te)|vk(40|5[0-3]|\-v)|vm40|voda|vulc|vx(52|53|60|61|70|80|81|83|85|98)|w3c(\-| )|webc|whit|wi(g |nc|nw)|wmlb|wonu|x700|yas\-|your|zeto|zte\-/i.test(a.substr(0,4)))check = true})(navigator.userAgent||navigator.vendor||window.opera); return check; } function display_stats(){ $("#article_stats_div").toggle(); return false; } /* * Cited By Scopus */ function citedCount(){ $("#framed_div_cited_count").toggle('fast', function(){ if ($(this).css('display') != 'none') { var loaded = document.getElementById("loaded_cite_count"); // to load only once the result! if(loaded.innerHTML == "No") { // Load Xref result var d = document.getElementById("framed_div_cited_count"); // This replace the content d.innerHTML = "<img src=\"https://pub.mdpi-res.com/img/loading_circle.gif?9a82694213036313?1732615622\" height=\"20\" width=\"20\" alt=\"Processing...\" style=\"vertical-align:middle; margin-right:0.6em;\">"; $.ajax({ method : "POST", url : "/cite-count/10.3390%252Fbiomedicines12102216", success : function(data) { if (data.succ) { d.innerHTML = data.view; loaded.innerHTML = "Yes"; follow_goto(); } } }); } } // end else return true; // for not going at the beginning of the page... }) return true; // for not going at the beginning of the page... } </script><script type="text/javascript" src="https://pub.mdpi-res.com/assets/js/third-party/highcharts/highcharts.js?bdd06f45e34c33df?1732615622"></script><script type="text/javascript" src="https://pub.mdpi-res.com/assets/js/third-party/highcharts/modules/exporting.js?944dc938d06de3a8?1732615622"></script><script type="text/javascript" defer="defer"> var advancedStatsData; var selectedStatsType = "abstract"; $(function(){ var countWrapper = $('#counts-wrapper'); $('#author_stats_id #type_links a').on('click', function(e) { e.preventDefault(); selectedStatsType = $(this).data('type'); $('#article_advanced_stats').vectorMap('set', 'values', advancedStatsData[selectedStatsType]); $('#advanced_stats_max').html(advancedStatsData[selectedStatsType].max); $('#type_links a').removeClass('active'); $(this).addClass('active'); }); $.get('/2227-9059/12/10/2216/stats', function (result) { if (!result.success) { return; } // process article metrics part in left column var viewNumber = countWrapper.find(".view-number"); viewNumber.html(result.metrics.views); viewNumber.parent().toggleClass("count-div--grey", result.metrics.views == 0); var downloadNumber = countWrapper.find(".download-number"); downloadNumber.html(result.metrics.downloads); downloadNumber.parent().toggleClass("count-div--grey", result.metrics.downloads == 0); var citationsNumber = countWrapper.find(".citations-number"); citationsNumber.html(result.metrics.citations); citationsNumber.parent().toggleClass("count-div--grey", result.metrics.citations == 0); if (result.metrics.views > 0 || result.metrics.downloads > 0 || result.metrics.citations > 0) { countWrapper.find("#js-counts-wrapper__views, #js-counts-wrapper__downloads").addClass("visible").show(); if (result.metrics.citations > 0) { countWrapper.find('.citations-number').html(result.metrics.citations).show(); countWrapper.find("#js-counts-wrapper__citations").addClass("visible").show(); } else { countWrapper.find("#js-counts-wrapper__citations").remove(); } $("[data-id='article-counters']").removeClass("hidden"); } if (result.metrics.altmetrics_score > 0) { $("#js-altmetrics-donut").show(); } // process view chart in main column var jsondata = result.chart; var series = new Array(); $.each(jsondata.elements, function(i, element) { var dataValues = new Array(); $.each(element.values, function(i, value) { dataValues.push(new Array(value.tip, value.value)); }); series[i] = {name: element.text, data:dataValues}; }); Highcharts.setOptions({ chart: { style: { fontFamily: 'Arial,sans-serif' } } }); $('#article_stats_swf').highcharts({ chart: { type: 'line', width: $("#tabs").width() //* 0.91 }, credits: { enabled: false }, exporting: { enabled: true }, title: { text: jsondata.title.text, x: -20 //center }, xAxis: { categories: jsondata.x_axis.labels.labels, offset: jsondata.x_axis.offset, labels:{ step: jsondata.x_axis.labels.steps, rotation: 30 } }, yAxis: { max: jsondata.y_axis.max, min: jsondata.y_axis.min, offset: jsondata.y_axis.offset, labels: { steps: jsondata.y_axis.steps }, title: { enabled: false } }, tooltip: { formatter: function (){ return this.key.replace("#val#", this.y); } }, legend: { align: 'top', itemDistance: 50 }, series: series }); }); $('#supplement_link').click(function() { document.getElementById('suppl_id').scrollIntoView(); }); $('#stats_link').click(function() { document.getElementById('stats_id').scrollIntoView(); }); // open mol viewer for molbank special supplementary files $('.showJmol').click(function(e) { e.preventDefault(); var jmolModal = $("#jmolModal"); var url = "/article/1488137/jsmol_viewer/__supplementary_id__"; url = url.replace(/__supplementary_id__/g, $(this).data('index')); $('#jsmol-content').attr('src', url); jmolModal.find(".content").html($(this).data('description')); jmolModal.foundation("reveal", "open"); }); }); !function() { "use strict"; function e(e) { try { if ("undefined" == typeof console) return; "error"in console ? console.error(e) : console.log(e) } catch (e) {} } function t(e) { return d.innerHTML = '<a href="' + e.replace(/"/g, """) + '"></a>', d.childNodes[0].getAttribute("href") || "" } function n(n, c) { var o = ""; var k = parseInt(n.substr(c + 4, 2), 16); for (var i = c; i < n.length; i += 2) { if (i != c + 4) { var s = parseInt(n.substr(i, 2), 16) ^ k; o += String.fromCharCode(s); } } try { o = decodeURIComponent(escape(o)); } catch (error) { console.error(error); } return t(o); } function c(t) { for (var r = t.querySelectorAll("a"), c = 0; c < r.length; c++) try { var o = r[c] , a = o.href.indexOf(l); a > -1 && (o.href = "mailto:" + n(o.href, a + l.length)) } catch (i) { e(i) } } function o(t) { for (var r = t.querySelectorAll(u), c = 0; c < r.length; c++) try { var o = r[c] , a = o.parentNode , i = o.getAttribute(f); if (i) { var l = n(i, 0) , d = document.createTextNode(l); a.replaceChild(d, o) } } catch (h) { e(h) } } function a(t) { for (var r = t.querySelectorAll("template"), n = 0; n < r.length; n++) try { i(r[n].content) } catch (c) { e(c) } } function i(t) { try { c(t), o(t), a(t) } catch (r) { e(r) } } var l = "/cnd-cgi/l/email-protection#" , u = ".__cf_email__" , f = "data-cfemail" , d = document.createElement("div"); i(document), function() { var e = document.currentScript || document.scripts[document.scripts.length - 1]; e.parentNode.removeChild(e) }() }(); </script><script type="text/javascript"> function setCookie(cname, cvalue, ctime) { ctime = (typeof ctime === 'undefined') ? 10*365*24*60*60*1000 : ctime; // default => 10 years var d = new Date(); d.setTime(d.getTime() + ctime); // ==> 1 hour = 60*60*1000 var expires = "expires="+d.toUTCString(); document.cookie = cname + "=" + cvalue + "; " + expires +"; path=/"; } function getCookie(cname) { var name = cname + "="; var ca = document.cookie.split(';'); for(var i=0; i<ca.length; i++) { var c = ca[i]; while (c.charAt(0)==' ') c = c.substring(1); if (c.indexOf(name) == 0) return c.substring(name.length, c.length); } return ""; } </script><script type="text/javascript" src="https://d1bxh8uas1mnw7.cloudfront.net/assets/embed.js"></script><script> $(document).ready(function() { if ($("#js-similarity-related-data").length > 0) { $.ajax({ url: '/article/1488137/similarity-related', success: function(response) { $("#js-similarity-related-data").html(response); $("#js-related-articles-menu").show(); $(document).foundation('tab', 'reflow'); MathJax.Hub.Queue(["Typeset", MathJax.Hub]); } }); } }); </script><link rel="stylesheet" href="https://pub.mdpi-res.com/assets/css/jquery-ui-1.10.4.custom.min.css?80647d88647bf347?1732615622"><link rel="stylesheet" href="https://pub.mdpi-res.com/assets/css/magnific-popup.min.css?04d343e036f8eecd?1732615622"><script type="text/javascript" src="https://pub.mdpi-res.com/assets/js/magnific-popup.min.js?2be3d9e7dc569146?1732615622"></script><script> $(function() { $(".js-show-more-academic-editors").on("click", function(e) { e.preventDefault(); $(this).hide(); $(".academic-editor-container").removeClass("hidden"); }); }); </script> <link rel="stylesheet" href="https://pub.mdpi-res.com/assets/css/vmap/jqvmap.min.css?126a06688aa11c13?1732615622"> <script src="https://pub.mdpi-res.com/assets/js/vmap/jquery.vmap.min.js?935f68d33bdd88a1?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/vmap/jquery.vmap.world.js?16677403c0e1bef1?1732615622"></script> <script> function updateSlick() { $('.multiple-items').slick('setPosition'); } $(document).ready(function() { $('.multiple-items').slick({ slidesToShow: 1, nextArrow: '<a class="slick-next" href="#"><i class="material-icons">chevron_right</i></a>', prevArrow: '<a class="slick-prev" href="#"><i class="material-icons">chevron_left</i></a>', slidesToScroll: 1, responsive: [ { breakpoint: 1024, settings: { slidesToShow: 1, slidesToScroll: 1, } }, { breakpoint: 600, settings: { slidesToShow: 1, slidesToScroll: 1, } }, { breakpoint: 480, settings: { slidesToShow: 1, slidesToScroll: 1, } } ] }); $('.multiple-items').show(); $(document).on('click', '.reviewReportSelector', function(e) { let path = $(this).attr('data-path'); handleReviews(path, $(this)); }); $(document).on('click', '.viewReviewReports', function(e) { let versionOne = $('#versionTab_1'); if (!versionOne.hasClass('activeTab')) { let path = $(this).attr('data-path'); handleReviews(path, versionOne); } location.href = "#reviewReports"; }); $(document).on('click', '.reviewersResponse, .authorResponse', function(e) { let version = $(this).attr('data-version'); let targetVersion = $('#versionTab_' + version); if (!targetVersion.hasClass('activeTab')) { let path = targetVersion.attr('data-path'); handleReviews(path, targetVersion); } location.href = $(this).attr('data-link'); }); $(document).on('click', '.tab', function (e) { e.preventDefault(); $('.tab').removeClass('activeTab'); $(this).addClass('activeTab') $('.tab').each(function() { $(this).closest('.tab-title').removeClass('active'); }); $(this).closest('.tab-title').addClass('active') }); }); function handleReviews(path, target) { $.ajax({ url: path, context: this, success: function (data) { $('.activeTab').removeClass('activeTab'); target.addClass('activeTab'); $('#reviewSection').html(data.view); }, error: function (xhr, ajaxOptions, thrownError) { console.log(xhr.status); console.log(thrownError); } }); } </script> <script src="https://pub.mdpi-res.com/assets/js/xmltohtml/affix.js?v1?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/xmltohtml/storage.js?e9b262d3a3476d25?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/xmltohtml/jquery-scrollspy.js?09cbaec0dbb35a67?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/xmltohtml/magnific-popup.js?4a09c18460afb26c?1732615622"></script> <script src="https://pub.mdpi-res.com/assets/js/xmltohtml/underscore.js?f893e294cde60c24?1732615622"></script> <script type="text/javascript"> $('document').ready(function(){ $("#left-column").addClass("show-for-large-up"); $("#middle-column").removeClass("medium-9").removeClass("left-bordered").addClass("medium-12"); $(window).on('resize scroll', function() { /* if ($('.button--drop-down').isInViewport($(".top-bar").outerHeight())) { */ if ($('.button--drop-down').isInViewport()) { $("#js-button-download").hide(); } else { $("#js-button-download").show(); } }); }); $(document).on('DOMNodeInserted', function(e) { var element = $(e.target); if (element.hasClass('menu') && element.hasClass('html-nav') ) { element.addClass("side-menu-ul"); } }); </script> <script src="https://pub.mdpi-res.com/assets/js/xmltohtml/articles.js?5118449d9ad8913a?1732615622"></script> <script> repositionOpenSideBar = function() { $('#left-column').addClass("show-for-large-up show-for-medium-up").show(); $('#middle-column').removeClass('large-12').removeClass('medium-12'); $('#middle-column').addClass('large-9'); } repositionCloseSideBar = function() { $('#left-column').removeClass("show-for-large-up show-for-medium-up").hide(); $('#middle-column').removeClass('large-9'); $('#middle-column').addClass('large-12').addClass('medium-12'); } </script>   <script type="text/plain" data-cookieconsent="marketing"> !function(e,t,n,s,u,a){e.twq||(s=e.twq=function(){s.exe?s.exe.apply(s,arguments):s.queue.push(arguments); },s.version='1.1',s.queue=[],u=t.createElement(n),u.async=!0,u.src='//static.ads-twitter.com/uwt.js', a=t.getElementsByTagName(n)[0],a.parentNode.insertBefore(u,a))}(window,document,'script'); // Insert Twitter Pixel ID and Standard Event data below twq('init','o2pip'); twq('track','PageView'); </script>  <script>(function(){function c(){var b=a.contentDocument||a.contentWindow.document;if(b){var d=b.createElement('script');d.innerHTML="window.__CF$cv$params={r:'8e8e2de85af24834',t:'MTczMjY2OTIwNi4wMDAwMDA='};var a=document.createElement('script');a.nonce='';a.src='/cdn-cgi/challenge-platform/scripts/jsd/main.js';document.getElementsByTagName('head')[0].appendChild(a);";b.getElementsByTagName('head')[0].appendChild(d)}}if(document.body){var a=document.createElement('iframe');a.height=1;a.width=1;a.style.position='absolute';a.style.top=0;a.style.left=0;a.style.border='none';a.style.visibility='hidden';document.body.appendChild(a);if('loading'!==document.readyState)c();else if(window.addEventListener)document.addEventListener('DOMContentLoaded',c);else{var e=document.onreadystatechange||function(){};document.onreadystatechange=function(b){e(b);'loading'!==document.readyState&&(document.onreadystatechange=e,c())}}}})();</script></body> </html>

CINXE.COM

Kalata B1 Enhances Temozolomide Toxicity to Glioblastoma Cells