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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 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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" > 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<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 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<label> Jordan </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_MALAYSIA"> <i class="material-icons">remove_circle_outline</i> </a> <label> Malaysia </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_UNITED_ARAB_EMIRATES"> <i class="material-icons">remove_circle_outline</i> </a> <label> United Arab Emirates </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_ARMENIA"> <i class="material-icons">remove_circle_outline</i> </a> <label> Armenia </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_AUSTRIA"> <i class="material-icons">remove_circle_outline</i> </a> <label> Austria </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_CHINA"> <i class="material-icons">remove_circle_outline</i> </a> <label> China </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_CROATIA"> <i class="material-icons">remove_circle_outline</i> </a> <label> Croatia </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_CZECH_REPUBLIC"> <i class="material-icons">remove_circle_outline</i> </a> <label> Czech Republic </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_DENMARK"> <i class="material-icons">remove_circle_outline</i> </a> <label> Denmark </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_EGYPT"> <i class="material-icons">remove_circle_outline</i> </a> <label> Egypt </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_PUERTO_RICO"> <i class="material-icons">remove_circle_outline</i> </a> <label> Puerto Rico </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_SOUTH_AFRICA"> <i class="material-icons">remove_circle_outline</i> </a> <label> South Africa </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_THAILAND"> <i class="material-icons">remove_circle_outline</i> </a> <label> Thailand </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_TUNISIA"> <i class="material-icons">remove_circle_outline</i> </a> <label> Tunisia </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_URUGUAY"> <i class="material-icons">remove_circle_outline</i> </a> <label> Uruguay </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_ARGENTINA"> <i class="material-icons">remove_circle_outline</i> </a> <label> Argentina </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_BELARUS"> <i class="material-icons">remove_circle_outline</i> </a> <label> Belarus </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_BELGIUM"> <i class="material-icons">remove_circle_outline</i> </a> <label> Belgium </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_CUBA"> <i class="material-icons">remove_circle_outline</i> </a> <label> Cuba </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_CYPRUS"> <i class="material-icons">remove_circle_outline</i> </a> <label> Cyprus </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_INDONESIA"> <i class="material-icons">remove_circle_outline</i> </a> <label> Indonesia </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_IRAQ"> <i class="material-icons">remove_circle_outline</i> </a> <label> Iraq </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_KAZAKHSTAN"> <i class="material-icons">remove_circle_outline</i> </a> <label> Kazakhstan </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_LEBANON"> <i class="material-icons">remove_circle_outline</i> </a> <label> Lebanon </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_MALAWI"> <i class="material-icons">remove_circle_outline</i> </a> <label> Malawi </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_MOROCCO"> <i class="material-icons">remove_circle_outline</i> </a> <label> Morocco </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_NIGERIA"> <i class="material-icons">remove_circle_outline</i> </a> <label> Nigeria </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_NORWAY"> <i class="material-icons">remove_circle_outline</i> </a> <label> Norway </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_PAKISTAN"> <i class="material-icons">remove_circle_outline</i> </a> <label> Pakistan </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_PHILIPPINES"> <i class="material-icons">remove_circle_outline</i> </a> <label> Philippines </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_SLOVAKIA"> <i class="material-icons">remove_circle_outline</i> </a> <label> Slovakia </label> </div/> <div class="remove-filter-container remove-filter-container--hidden"> <a href="#" class="remove-filter-link link--red " data-filterid="refine_countries_SWEDEN"> <i class="material-icons">remove_circle_outline</i> </a> <label> Sweden 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href="/2673-6284/13/4/51/pdf?version=1732185868" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="The Biotechnological Potential of Crickets as a Sustainable Protein Source for Fishmeal Replacement in Aquafeed" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <div class="article-icons"><span class="label openaccess" data-dropdown="drop-article-label-openaccess" aria-expanded="false">Open Access</span><span class="label articletype">Review</span></div> <a class="title-link" href="/2673-6284/13/4/51">The Biotechnological Potential of Crickets as a Sustainable Protein Source for Fishmeal Replacement in Aquafeed</a> <div class="authors"> by <span class="inlineblock "><strong>Aldo Fraijo-Valenzuela</strong>, </span><span class="inlineblock "><strong>Joe Luis Arias-Moscoso</strong>, </span><span class="inlineblock "><strong>Oscar Daniel García-Pérez</strong>, </span><span class="inlineblock "><strong>Libia Zulema Rodriguez-Anaya</strong> and </span><span class="inlineblock "><strong>Jose Reyes Gonzalez-Galaviz</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 51; <a href="https://doi.org/10.3390/biotech13040051">https://doi.org/10.3390/biotech13040051</a> - 21 Nov 2024 </div> Viewed by 305 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> As aquaculture production grows, so does the demand for quality and cost-effective protein sources. The cost of fishmeal (FM) has increased over the years, leading to increased production costs for formulated aquafeed. Soybean meal (SBM) is commonly used as an FM replacer in <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/51/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> As aquaculture production grows, so does the demand for quality and cost-effective protein sources. The cost of fishmeal (FM) has increased over the years, leading to increased production costs for formulated aquafeed. Soybean meal (SBM) is commonly used as an FM replacer in aquafeed, but anti-nutritional factors could affect the growth, nutrition, and health of aquatic organisms. Cricket meal (CM) is an alternative source with a nutrient profile comparable to FM due to its high protein content, digestibility, and amino acid profile. CM use in aquafeed influences growth and reproductive performance while modulating the gut microbiota and immune response of fish and shrimp. However, consistent regulation and scaling up are necessary for competitive prices and the marketing of CM. Moreover, the chitin content in CM could be an issue in some fish species; however, different strategies based on food biotechnology can improve the protein quality for its safe use in aquafeed. <a href="/2673-6284/13/4/51">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Section <a href="/journal/biotech/sections/agricultural_food_biotechnology">Agricultural and Food Biotechnology</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/51/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1526105"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1526105"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1526105" data-cycle-prev="#prev1526105" data-cycle-progressive="#images1526105" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1526105-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00051/article_deploy/html/images/biotech-13-00051-g001-550.jpg?1732185949" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1526105" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1526105-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00051/article_deploy/html/images/biotech-13-00051-g002-550.jpg?1732185951'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1526105-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00051/article_deploy/html/images/biotech-13-00051-g003-550.jpg?1732185952'><p>Figure 3</p></div></script></div></div><div id="article-1526105-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00051/article_deploy/html/images/biotech-13-00051-g001-550.jpg?1732185949" title=" <strong>Figure 1</strong><br/> <p>Nutritional composition of cricket meal. Created with BioRender.com (accessed on 9 October 2024).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/51'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00051/article_deploy/html/images/biotech-13-00051-g002-550.jpg?1732185951" title=" <strong>Figure 2</strong><br/> <p>Illustration of cricket meal’s effects on aquaculture. Created with BioRender.com (accessed on 9 October 2024).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/51'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00051/article_deploy/html/images/biotech-13-00051-g003-550.jpg?1732185952" title=" <strong>Figure 3</strong><br/> <p>Illustration of different processing methods to improve protein quality of cricket meal. Created with BioRender.com (accessed on 9 October 2024).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/51'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1521729" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 14 pages, 2500 KiB   </span> <a href="/2673-6284/13/4/50/pdf?version=1731667598" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Leveraging Walnut Somatic Embryos as a Biomanufacturing Platform for Recombinant Proteins and Metabolites" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <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> <a class="title-link" href="/2673-6284/13/4/50">Leveraging Walnut Somatic Embryos as a Biomanufacturing Platform for Recombinant Proteins and Metabolites</a> <div class="authors"> by <span class="inlineblock "><strong>Paulo A. Zaini</strong>, </span><span class="inlineblock "><strong>Katherine R. Haddad</strong>, </span><span class="inlineblock "><strong>Noah G. Feinberg</strong>, </span><span class="inlineblock "><strong>Yakir Ophir</strong>, </span><span class="inlineblock "><strong>Somen Nandi</strong>, </span><span class="inlineblock "><strong>Karen A. McDonald</strong> and </span><span class="inlineblock "><strong>Abhaya M. Dandekar</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 50; <a href="https://doi.org/10.3390/biotech13040050">https://doi.org/10.3390/biotech13040050</a> - 15 Nov 2024 </div> Viewed by 396 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Biomanufacturing enables novel sources of compounds with constant demand, such as food coloring and preservatives, as well as new compounds with peak demand, such as diagnostics and vaccines. The COVID-19 pandemic has highlighted the need for alternative sources of research materials, thrusting research <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/50/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Biomanufacturing enables novel sources of compounds with constant demand, such as food coloring and preservatives, as well as new compounds with peak demand, such as diagnostics and vaccines. The COVID-19 pandemic has highlighted the need for alternative sources of research materials, thrusting research on diversification of biomanufacturing platforms. Here, we show initial results exploring the walnut somatic embryogenic system expressing the recombinant receptor binding domain (RBD) and ectodomain of the spike protein (Spike) from the SARS-CoV-2 virus. Stably transformed walnut embryo lines were selected and propagated in vitro. Both recombinant proteins were detected at 3–14 µg/g dry weight of tissue culture material. Although higher yields of recombinant protein have been obtained using more conventional biomanufacturing platforms, we also report on the production of the red pigment betanin in somatic embryos, reaching yields of 650 mg/g, even higher than red beet <i>Beta vulgaris</i>. This first iteration shows the potential of biomanufacturing using somatic walnut embryos that can now be further optimized for different applications sourcing specialized proteins and metabolites. <a href="/2673-6284/13/4/50">Full article</a> </div> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/50/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1521729"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1521729"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1521729" data-cycle-prev="#prev1521729" data-cycle-progressive="#images1521729" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1521729-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00050/article_deploy/html/images/biotech-13-00050-g001-550.jpg?1731667678" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1521729" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1521729-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00050/article_deploy/html/images/biotech-13-00050-g002-550.jpg?1731667680'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1521729-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00050/article_deploy/html/images/biotech-13-00050-g003-550.jpg?1731667682'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1521729-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00050/article_deploy/html/images/biotech-13-00050-g004-550.jpg?1731667683'><p>Figure 4</p></div></script></div></div><div id="article-1521729-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00050/article_deploy/html/images/biotech-13-00050-g001-550.jpg?1731667678" title=" <strong>Figure 1</strong><br/> <p>Diagram showing plant expression vectors used in this study. (<b>A</b>) Two sets of vectors were tested: one derived from Addgene vector 160908 expressing the red pigment betanin encoded by RUBY and the other derived from Takara vector pRI 201-AN with inserted geminiviral components for enhanced DNA replication [<a href="#B30-biotech-13-00050" class="html-bibr">30</a>,<a href="#B31-biotech-13-00050" class="html-bibr">31</a>]. In each vector type, both RBD (aa. 331-521 of full-length spike in pRUBY-RBD and aa. 319-541 in pGEMINI-RBD) and SPIKE ectodomain (aa. 36-1167 and 16-1209, respectively) of SARS-CoV-2 spike protein were encoded with a C-terminal HisTag. For RUBY vectors, variations in the expression cassette were tested for enhanced expression. This included no 5′-UTR sequence, SpeI restriction site, or the 5′-UTR present in the Gemini vector (from the <span class="html-italic">Arabidopsis thaliana</span> alcohol dehydrogenase ADH). (<b>B</b>) Sequences of recombinant proteins are shown for RBD and SPIKE. The sequences present only in pGEMINI are shown in bold, and the sites underlined are substituted with prolines in the SPIKE protein expressed in pRUBY.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/50'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00050/article_deploy/html/images/biotech-13-00050-g002-550.jpg?1731667680" title=" <strong>Figure 2</strong><br/> <p>Growth of walnut embryos on solid and liquid media. (<b>A</b>) E2 embryo lines growing well on solid medium (in 100 mm Petri dishes) were monitored for biomass increase: pGEMINI-RBD is represented by line GR03, and pRUBY-RBD is represented by line SA05. (<b>B</b>) Growth of selected lines observed in liquid DKW medium. Values shown are averages ± standard deviation of three independent flasks.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/50'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00050/article_deploy/html/images/biotech-13-00050-g003-550.jpg?1731667682" title=" <strong>Figure 3</strong><br/> <p>Expression of RBD and SPIKE proteins in selected walnut embryo lines. Quantification of recombinant protein in soluble protein extracts by ELISA using anti-HisTag—HRP—conjugated antibody diluted 1:1000. Values shown are averages ± standard deviation of two independent experiments with two replicates each. Walnut embryo lines with G in the identifier are derived from Gemini vectors, and those with S are derived from RUBY vectors. SA and SS lines express RBD as well as GR, while GS lines express Spike. J1 WT was used as a control for pGEMINI-expressing lines, and pRUBY empty vector (EV) was used as a control for pRUBY-expressing lines.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/50'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00050/article_deploy/html/images/biotech-13-00050-g004-550.jpg?1731667683" title=" <strong>Figure 4</strong><br/> <p>Quantification of betanin produced in walnut somatic embryos. Non-pigmented GR03 walnut embryos were compared with line SA05 expressing RUBY and with an extract prepared from red beet. Quantification was achieved using a calibration curve based on a serially diluted betanin chemical standard, with absorbance measurements taken at 531 nm (within the reported absorbance range for betanin). Spectrophotometric measurements were taken in triplicates from three biological replicates. Average ± standard deviation is shown. Difference between walnut SA05 and red beet considered significant by Dunn’s test (<span class="html-italic">p</span>-value &lt; 0.05) and between walnut SA05 and GR03 samples (<span class="html-italic">p</span>-value &lt; 0.001).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/50'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1521439" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 18 pages, 518 KiB   </span> <a href="/2673-6284/13/4/49/pdf?version=1731656635" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Climate Resilience in Farm Animals: Transcriptomics-Based Alterations in Differentially Expressed Genes and Stress Pathways" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <div class="article-icons"><span class="label openaccess" data-dropdown="drop-article-label-openaccess" aria-expanded="false">Open Access</span><span class="label articletype">Review</span></div> <a class="title-link" href="/2673-6284/13/4/49">Climate Resilience in Farm Animals: Transcriptomics-Based Alterations in Differentially Expressed Genes and Stress Pathways</a> <div class="authors"> by <span class="inlineblock "><strong>Chikamagalore Gopalakrishna Shashank</strong>, </span><span class="inlineblock "><strong>Veerasamy Sejian</strong>, </span><span class="inlineblock "><strong>Mullakkalparambil Velayudhan Silpa</strong>, </span><span class="inlineblock "><strong>Chinnasamy Devaraj</strong>, </span><span class="inlineblock "><strong>Aradotlu Parameshwarappa Madhusoodan</strong>, </span><span class="inlineblock "><strong>Ebenezer Binuni Rebez</strong>, </span><span class="inlineblock "><strong>Gajendirane Kalaignazhal</strong>, </span><span class="inlineblock "><strong>Artabandhu Sahoo</strong> and </span><span class="inlineblock "><strong>Frank Rowland Dunshea</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 49; <a href="https://doi.org/10.3390/biotech13040049">https://doi.org/10.3390/biotech13040049</a> - 15 Nov 2024 </div> Viewed by 391 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> The livestock sector, essential for maintaining food supply and security, encounters numerous obstacles as a result of climate change. Rising global populations exacerbate competition for natural resources, affecting feed quality and availability, heightening livestock disease risks, increasing heat stress, and contributing to biodiversity <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/49/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> The livestock sector, essential for maintaining food supply and security, encounters numerous obstacles as a result of climate change. Rising global populations exacerbate competition for natural resources, affecting feed quality and availability, heightening livestock disease risks, increasing heat stress, and contributing to biodiversity loss. Although various management and dietary interventions exist to alleviate these impacts, they often offer only short-lived solutions. We must take a more comprehensive approach to understanding how animals adapt to and endure their environments. One such approach is quantifying transcriptomes under different environments, which can uncover underlying pathways essential for livestock adaptation. This review explores the progress and techniques in studies that apply gene expression analysis to livestock production systems, focusing on their adaptation to climate change. We also attempt to identify various biomarkers and transcriptomic differences between species and pure/crossbred animals. Looking ahead, integrating emerging technologies such as spatialomics could further accelerate genetic improvements, enabling more thermoresilient and productive livestock in response to future climate fluctuations. Ultimately, insights from these studies will help optimize livestock production systems by identifying thermoresilient/desired animals for use in precise breeding programs to counter climate change. <a href="/2673-6284/13/4/49">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Section <a href="/journal/biotech/sections/agricultural_food_biotechnology">Agricultural and Food Biotechnology</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/49/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="absgraph cycle-slideshow"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1521439-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00049/article_deploy/html/images/biotech-13-00049-g001-550.jpg?1731656742" alt="" style="border: 0;"><p>Figure 1</p></div></div></div><div id="article-1521439-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00049/article_deploy/html/images/biotech-13-00049-g001-550.jpg?1731656742" title=" <strong>Figure 1</strong><br/> <p>Application of transcriptomic data analysis using Clusters of Orthologous Groups of proteins (COGs) and Kyoto Encyclopedia of Genes and Genomes (KEGG) database in identifying changes associated with heat stress.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/49'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1520621" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 16 pages, 2975 KiB   </span> <a href="/2673-6284/13/4/48/pdf?version=1731578277" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="High-Resolution Melting Analysis Potential for Saccharomyces cerevisiae var. boulardii Authentication in Probiotic-Enriched Food Matrices" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <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> <a class="title-link" href="/2673-6284/13/4/48">High-Resolution Melting Analysis Potential for <i>Saccharomyces cerevisiae</i> var. <i>boulardii</i> Authentication in Probiotic-Enriched Food Matrices</a> <div class="authors"> by <span class="inlineblock "><strong>Monika Borkowska</strong>, </span><span class="inlineblock "><strong>Michał Kułakowski</strong> and </span><span class="inlineblock "><strong>Kamila Myszka</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 48; <a href="https://doi.org/10.3390/biotech13040048">https://doi.org/10.3390/biotech13040048</a> - 14 Nov 2024 </div> Viewed by 327 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> To date, the only probiotic yeast with evidence of health-promoting effects is <i>Saccharomyces cerevisiae</i> var. <i>boulardii</i>. The expanded market including dietary supplements and functional foods supplemented with <i>Saccharomyces cerevisiae</i> var. <i>boulardii</i> creates an environment conductive to food adulterations, necessitating rapid testing to verify <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/48/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> To date, the only probiotic yeast with evidence of health-promoting effects is <i>Saccharomyces cerevisiae</i> var. <i>boulardii</i>. The expanded market including dietary supplements and functional foods supplemented with <i>Saccharomyces cerevisiae</i> var. <i>boulardii</i> creates an environment conductive to food adulterations, necessitating rapid testing to verify product probiotic status. Herein, qPCR-HRM analysis was tested for probiotic yeast identification. The effectiveness of the primer pairs’ set was examined, designed to amplify heterogeneous regions in (a) rDNA sequences previously designed to identify food-derived yeast and (b) genes associated with physiological and genotypic divergence of <i>Saccharomyces cerevisiae</i> var. <i>boulardii.</i> Preliminary tests of amplicons’ differentiation power enabled the selection of interspecies sequences for <i>18SrRNA</i> and ITS and genus-specific sequences <i>HO</i>, <i>RPB2</i>, <i>HXT9</i> and <i>MAL11.</i> The multi-fragment qPCR-HRM analysis was sufficient for culture-dependent <i>Saccharomyces cerevisiae</i> var. <i>boulardii</i> identification and proved effective in the authentication of dietary supplements’ probiotic composition. The identification of <i>S. cerevisiae</i> var. <i>boulardii</i> in complex microbial mixtures of kefir succeeded with more specific intragenus sequences <i>HO</i> and <i>RPB2.</i> The predominance of <i>S. cerevisiae</i> var. <i>boulardii</i> in the tested matrices, quantitatively corresponded to the probiotic-enriched food, was crucial for identification with qPCR–HRM analysis. Considering the reported assumptions, qPCR-HRM analysis is an appropriate tool for verifying probiotic-enriched food. <a href="/2673-6284/13/4/48">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Section <a href="/journal/biotech/sections/agricultural_food_biotechnology">Agricultural and Food Biotechnology</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/48/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1520621"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1520621"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1520621" data-cycle-prev="#prev1520621" data-cycle-progressive="#images1520621" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1520621-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00048/article_deploy/html/images/biotech-13-00048-g001-550.jpg?1731578384" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1520621" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1520621-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00048/article_deploy/html/images/biotech-13-00048-g002-550.jpg?1731578387'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1520621-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00048/article_deploy/html/images/biotech-13-00048-g003-550.jpg?1731578390'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1520621-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00048/article_deploy/html/images/biotech-13-00048-g004-550.jpg?1731578391'><p>Figure 4</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1520621-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00048/article_deploy/html/images/biotech-13-00048-g005-550.jpg?1731578394'><p>Figure 5</p></div></script></div></div><div id="article-1520621-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00048/article_deploy/html/images/biotech-13-00048-g001-550.jpg?1731578384" title=" <strong>Figure 1</strong><br/> <p>Verification of sporulation and HXT9 or MAL11 presence in Sc strains. Microscopic images of yeast cells of Sb745 (<b>a</b>), Sb3799 (<b>b</b>), ScPS1 (<b>c</b>), ScD (<b>d</b>) strains. The cells’ suspensions were induced to sporulate under starvation conditions and then stained with the Ziehl–Neelsen method. The arrows indicate stained ascospores (<b>c</b>,<b>d</b>). Electrophoretic separation of HXT9 (<b>e</b>) and MALL1 (<b>f</b>) amplicons obtained in PCR for <span class="html-italic">S. cerevisiae</span> var. <span class="html-italic">boulardii</span> reference strains (Sb745, Sb3799), <span class="html-italic">S. cerevisiae</span> strains (ScATCC9763, ScEtRed, ScD and ScPS1), <span class="html-italic">K. marxianus</span> (<span class="html-italic">Km</span>) and <span class="html-italic">P. fermentans</span> (<span class="html-italic">Pf</span>). M—DNA Marker 100 bp LOAD (Syngen Biotech, Wroclaw, Poland).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/48'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00048/article_deploy/html/images/biotech-13-00048-g002-550.jpg?1731578387" title=" <strong>Figure 2</strong><br/> <p>Differentiation of yeast with interspecies primer pairs in qPCR-HRM analysis. Melt peaks (<b>a</b>) and difference curves grouped as color-marked clusters (<b>b</b>) were obtained by qPCR-HRM analysis of 18SrDNA, ITS, 26SrDNA and TEF1alpha regions. DNA templates of <span class="html-italic">S. cerevisiae</span> var. <span class="html-italic">boulardii</span> reference strains (Sb745, Sb3799), <span class="html-italic">S. cerevisiae</span> strains (ScATCC9763, ScEtRed, ScD), <span class="html-italic">K. marxianus</span> (Km) and <span class="html-italic">P. fermentans</span> (PfD) strains were amplified in technical duplicate.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/48'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00048/article_deploy/html/images/biotech-13-00048-g003-550.jpg?1731578390" title=" <strong>Figure 3</strong><br/> <p>Differentiation of Sc strains using intragenus primer pairs in qPCR-HRM analysis. Melt peaks (<b>a</b>) and difference curves grouped as color-marked clusters (<b>b</b>) were obtained by qPCR-HRM analysis of CCA1, HO, RPB2, MAL11and HXT9 regions. DNA templates of <span class="html-italic">S. cerevisiae</span> var. <span class="html-italic">boulardii</span> reference strains (Sb745, Sb3799), <span class="html-italic">S. cerevisiae</span> strains (ScATCC9763, ScEtRed, ScD), <span class="html-italic">K. marxianus</span> (Km) and <span class="html-italic">P. fermentans</span> (PfD) strains were amplified in technical duplicate. NA—not amplified.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/48'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00048/article_deploy/html/images/biotech-13-00048-g004-550.jpg?1731578391" title=" <strong>Figure 4</strong><br/> <p>Identification of Sb in dietary supplements with qPCR-HRM. Melt peaks (<b>a</b>) and difference curves grouped as color-marked clusters (<b>b</b>) were obtained by qPCR-HRM analysis of 18SrDNA and ITS regions (1), HO and RPB2 regions (2). DNA template of <span class="html-italic">S. cerevisiae</span> var. <span class="html-italic">boulardii</span> reference strain (Sb745) as positive control, <span class="html-italic">S. cerevisiae</span> reference (ScATCC9763) and the dietary supplements’ DNA templates (PS1, PS2, PS3, PS4) were amplified in technical duplicate.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/48'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00048/article_deploy/html/images/biotech-13-00048-g005-550.jpg?1731578394" title=" <strong>Figure 5</strong><br/> <p>Identification of Sb745 in microbial mixtures with qPCR-based HRM analysis. Melt peaks (<b>a</b>) and difference curves grouped as color-marked clusters (<b>b</b>) were obtained by qPCR-HRM analysis of 18SrDNA. Melt peaks (<b>c</b>)—the arrows indicate species-specific peaks at T<sub>m</sub> 81.6 °C, 82.2 °C and 84.4 °C for Km, ScD and PfD, respectively, and difference curves (<b>d</b>) grouped as color-marked clusters detected for ITS amplicon. Difference curves grouped as color-marked clusters obtained by qPCR-HRM analysis of HO sequence presented for all samples (<b>e</b>) and selected samples (<b>f</b>), and RPB2 sequence presented for all samples (<b>g</b>) and selected samples (<b>h</b>). DNA templates of <span class="html-italic">S. cerevisiae</span> var. <span class="html-italic">boulardii</span> reference strain (Sb745) as positive control and the microbial mixtures’ DNA extracts (Mx_Sb, Mx_0.9, Mx_0.5, Mx_0.1, Mx_Sc, S_0.9, S_0.5, S_0.1) were amplified in technical duplicate.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/48'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1520470" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 18 pages, 3391 KiB   </span> <a href="/2673-6284/13/4/47/pdf?version=1731564835" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Honey Bioactive Molecules: There Is a World Beyond the Sugars" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <div class="article-icons"><span class="label openaccess" data-dropdown="drop-article-label-openaccess" aria-expanded="false">Open Access</span><span class="label articletype">Review</span></div> <a class="title-link" href="/2673-6284/13/4/47">Honey Bioactive Molecules: There Is a World Beyond the Sugars</a> <div class="authors"> by <span class="inlineblock "><strong>Gregorio Bonsignore</strong>, </span><span class="inlineblock "><strong>Simona Martinotti</strong> and </span><span class="inlineblock "><strong>Elia Ranzato</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 47; <a href="https://doi.org/10.3390/biotech13040047">https://doi.org/10.3390/biotech13040047</a> - 14 Nov 2024 </div> Viewed by 368 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Honey’s many bioactive compounds have been utilized historically to cure infectious diseases. Beneficial effects are its antiviral, antibacterial, anti-inflammatory, antioxidant, and immune-stimulating qualities. The bee species, geographic location, botanical origin, harvest season, processing, and storage conditions all affect honey’s potential for therapeutic use. <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/47/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Honey’s many bioactive compounds have been utilized historically to cure infectious diseases. Beneficial effects are its antiviral, antibacterial, anti-inflammatory, antioxidant, and immune-stimulating qualities. The bee species, geographic location, botanical origin, harvest season, processing, and storage conditions all affect honey’s potential for therapeutic use. Honey contains a number of antioxidants and active compounds, such as polyphenols, which have been shown to have disease-preventive properties. Based on their origins, categories, and functions, the main polyphenols found in various honey varieties are examined in this review. <a href="/2673-6284/13/4/47">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Special Issue <a href=" /journal/biotech/special_issues/7KP0712M2L ">Natural Antioxidants: Determination in Food and Nutraceuticals and Implications on Human Health</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/47/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="absgraph cycle-slideshow"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1520470-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00047/article_deploy/html/images/biotech-13-00047-g001-550.jpg?1731564905" alt="" style="border: 0;"><p>Figure 1</p></div></div></div><div id="article-1520470-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00047/article_deploy/html/images/biotech-13-00047-g001-550.jpg?1731564905" title=" <strong>Figure 1</strong><br/> <p>Main recognized properties of honey. For more information, see the text. Created in BioRender. <a href="https://BioRender.com/s21r904" target="_blank">https://BioRender.com/s21r904</a> (accessed on 11 November 2024).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/47'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1518955" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 17 pages, 5204 KiB   </span> <a href="/2673-6284/13/4/46/pdf?version=1731401779" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Screening and Characterization of Sialic Acid-Binding Variable Lymphocyte Receptors from Hagfish" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <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> <a class="title-link" href="/2673-6284/13/4/46">Screening and Characterization of Sialic Acid-Binding Variable Lymphocyte Receptors from Hagfish</a> <div class="authors"> by <span class="inlineblock "><strong>Mark Rickard N. Angelia</strong>, </span><span class="inlineblock "><strong>Abigail Joy D. Rodelas-Angelia</strong>, </span><span class="inlineblock "><strong>Cheolung Yang</strong>, </span><span class="inlineblock "><strong>Sojeong Park</strong>, </span><span class="inlineblock "><strong>Seung pyo Jeong</strong>, </span><span class="inlineblock "><strong>Hyeok Jang</strong>, </span><span class="inlineblock "><strong>Dennis Berbulla Bela-ong</strong>, </span><span class="inlineblock "><strong>Hobin Jang</strong>, </span><span class="inlineblock "><strong>Kim D. Thompson</strong> and </span><span class="inlineblock "><strong>Taesung Jung</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 46; <a href="https://doi.org/10.3390/biotech13040046">https://doi.org/10.3390/biotech13040046</a> - 12 Nov 2024 </div> Viewed by 461 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Sialic acid is a diverse group of monosaccharides often found on the termini of <i>N</i>- and <i>O</i>-linked glycans as well as being components of glycoconjugates. Hypersialylation has been associated with the progression of chronic inflammation-mediated diseases such as cardiovascular disease and <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/46/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Sialic acid is a diverse group of monosaccharides often found on the termini of <i>N</i>- and <i>O</i>-linked glycans as well as being components of glycoconjugates. Hypersialylation has been associated with the progression of chronic inflammation-mediated diseases such as cardiovascular disease and cancer. Given its role in infection and disease-related processes, sialic acid is a promising target for therapeutic approaches that utilize carbohydrate-binding molecules. In this study, we screened for sialic acid-recognizing variable lymphocyte receptors (VLRBs) or ccombodies from inshore hagfish (<i>Eptatretus burgeri</i>) using a synthetic Neu5Ac-glycoconjugate as an antigen in immunoassay. Resulting ccombodies, 2D8, 5G11, 4A1, and 5F8 were further characterized in terms of their binding activity and specificity. A competitive ELISA using free haptens showed strong inhibition using either <i>N</i>-acetylneuraminic acid (Neu5Ac) and <i>N</i>-glycolylneuraminic acid (Neu5Gc). The half-maximal inhibitory concentrations (IC<sub>50</sub>) for Neu5Ac ranged from 7.02 to 17.06 mM, with candidates 4A1 and 5G11 requiring the least and highest amounts, respectively. IC<sub>50</sub> values for Neu5Gc ranged from 8.12 to 13.91 mM, for 4A1 and 5G11, respectively. Candidate ccombodies also detected naturally occurring sialic acid from known sialoglycoproteins using a dot blot assay. Neu5Gc-5G11 and Neu5Ac-2D8 yielded the strongest and weakest docking interactions with affinity values of −5.9 kcal/mol and −4.9 kcal/mol, respectively. Hydrogen bonding and hydrophobic interactions were predicted to be the predominant noncovalent forces observed between the ccombodies and sialic acid. This study demonstrates that glycan-binding VLRBs from hagfish hold promise in augmenting the glycobiologists’ toolkit in investigating the roles of glycans in human and animal health and disease. <a href="/2673-6284/13/4/46">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Section <a href="/journal/biotech/sections/agricultural_food_biotechnology">Agricultural and Food Biotechnology</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/46/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1518955"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1518955"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1518955" data-cycle-prev="#prev1518955" data-cycle-progressive="#images1518955" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1518955-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g001-550.jpg?1731401962" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1518955" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1518955-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g002-550.jpg?1731401964'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1518955-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g003-550.jpg?1731401965'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1518955-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g004-550.jpg?1731401966'><p>Figure 4</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1518955-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g005-550.jpg?1731401969'><p>Figure 5</p></div> --- <div class='openpopupgallery' data-imgindex='5' data-target='article-1518955-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g006-550.jpg?1731401973'><p>Figure 6</p></div> --- <div class='openpopupgallery' data-imgindex='6' data-target='article-1518955-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g007a-550.jpg?1731401975'><p>Figure 7</p></div> --- <div class='openpopupgallery' data-imgindex='7' data-target='article-1518955-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g007b-550.jpg?1731401979'><p>Figure 7 Cont.</p></div> --- <div class='openpopupgallery' data-imgindex='8' data-target='article-1518955-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g007c-550.jpg?1731401983'><p>Figure 7 Cont.</p></div></script></div></div><div id="article-1518955-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g001-550.jpg?1731401962" title=" <strong>Figure 1</strong><br/> <p>(<b>A</b>) SDS-PAGE profile of BSA-CC-Neu5Ac conjugate, (<b>B</b>) PAS stain of BSA-CC-Neu5Ac. Lane 1—MW marker, 2—BSA, 3—BSA-CC-Neu5Ac, 4—HRP (PAS staining positive control).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/46'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g002-550.jpg?1731401964" title=" <strong>Figure 2</strong><br/> <p>Antibody titration of various ccombodies against BSA-CC-Neu5Ac. Values are the average of two determinations, with the standard deviation shown as error bars.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/46'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g003-550.jpg?1731401965" title=" <strong>Figure 3</strong><br/> <p>Competitive ELISA of ccombody candidates against (<b>A</b>) Neu5Ac and (<b>B</b>) Neu5Gc.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/46'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g004-550.jpg?1731401966" title=" <strong>Figure 4</strong><br/> <p>Western blot assays of BSA-CC-Neu5Ac using candidates (<b>A</b>) 2D8, (<b>B</b>) 5G11, (<b>C</b>) 6D2, and (<b>D</b>) 4A1. Lane 1—BSA, 2—BSA-CC-Neu5Ac.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/46'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g005-550.jpg?1731401969" title=" <strong>Figure 5</strong><br/> <p>Representative dot blot assay and percent area intensity of ccombody candidates 2D8, 4A1, 5G11, and 6D2 against known sialoglycoproteins. Percent area intensity for each dot was obtained using densitometric analysis.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/46'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g006-550.jpg?1731401973" title=" <strong>Figure 6</strong><br/> <p>Molecular docking of ccombody candidates (<b>A</b>) 2D8, (<b>B</b>) 5G11, (<b>C</b>) 6D2, and (<b>D</b>) 4A1 with (<b>1</b>) Neu5Ac and (<b>2</b>) Neu5Gc using AutoDock Vina. Protein structures are represented using a ribbon model (left) and a space-filling model (right) while the glycan ligand is represented by a stick model.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/46'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g007a-550.jpg?1731401975" title=" <strong>Figure 7</strong><br/> <p>Noncovalent interactions between ccombodies and sialic acids (<b>A</b>) Neu5Ac and (<b>B</b>) Neu5Gc. Amino acid residues involved in hydrogen bonding (green dashed lines) are colored green. Red spoked arcs are residues that exert hydrophobic interactions. (<b>C</b>) Multiple sequence alignment of sialic acid-binding ccombodies. Residues in gray are positioned in the LRR hypervariable region and predicted to interact with the ligand.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/46'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g007b-550.jpg?1731401979" title=" <strong>Figure 7 Cont.</strong><br/> <p>Noncovalent interactions between ccombodies and sialic acids (<b>A</b>) Neu5Ac and (<b>B</b>) Neu5Gc. Amino acid residues involved in hydrogen bonding (green dashed lines) are colored green. Red spoked arcs are residues that exert hydrophobic interactions. (<b>C</b>) Multiple sequence alignment of sialic acid-binding ccombodies. Residues in gray are positioned in the LRR hypervariable region and predicted to interact with the ligand.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/46'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00046/article_deploy/html/images/biotech-13-00046-g007c-550.jpg?1731401983" title=" <strong>Figure 7 Cont.</strong><br/> <p>Noncovalent interactions between ccombodies and sialic acids (<b>A</b>) Neu5Ac and (<b>B</b>) Neu5Gc. Amino acid residues involved in hydrogen bonding (green dashed lines) are colored green. Red spoked arcs are residues that exert hydrophobic interactions. (<b>C</b>) Multiple sequence alignment of sialic acid-binding ccombodies. Residues in gray are positioned in the LRR hypervariable region and predicted to interact with the ligand.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/46'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1518151" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 16 pages, 2544 KiB   </span> <a href="/2673-6284/13/4/45/pdf?version=1731379983" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Artificial Insemination as a Possible Convenient Tool to Acquire Genome-Edited Mice via In Vivo Fertilization with Engineered Sperm" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <div class="article-icons"><span class="label openaccess" data-dropdown="drop-article-label-openaccess" aria-expanded="false">Open Access</span><span class="label articletype">Review</span></div> <a class="title-link" href="/2673-6284/13/4/45">Artificial Insemination as a Possible Convenient Tool to Acquire Genome-Edited Mice via In Vivo Fertilization with Engineered Sperm</a> <div class="authors"> by <span class="inlineblock "><strong>Masahiro Sato</strong>, </span><span class="inlineblock "><strong>Emi Inada</strong>, </span><span class="inlineblock "><strong>Issei Saitoh</strong>, </span><span class="inlineblock "><strong>Kazunori Morohoshi</strong> and </span><span class="inlineblock "><strong>Shingo Nakamura</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 45; <a href="https://doi.org/10.3390/biotech13040045">https://doi.org/10.3390/biotech13040045</a> - 11 Nov 2024 </div> Viewed by 670 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Advances in genome editing technology have made it possible to create genome-edited (GE) animals, which are useful for identifying isolated genes and producing models of human diseases within a short period of time. The production of GE animals mainly relies on the gene <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/45/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Advances in genome editing technology have made it possible to create genome-edited (GE) animals, which are useful for identifying isolated genes and producing models of human diseases within a short period of time. The production of GE animals mainly relies on the gene manipulation of pre-implantation embryos, such as fertilized eggs and two-cell embryos, which can usually be achieved by the microinjection of nucleic acids, electroporation in the presence of nucleic acids, or infection with viral vectors, such as adeno-associated viruses. In contrast, GE animals can theoretically be generated by fertilizing ovulated oocytes with GE sperm. However, there are only a few reports showing the successful production of GE animals using GE sperm. Artificial insemination (AI) is an assisted reproduction technology based on the introduction of isolated sperm into the female reproductive tract, such as the uterine horn or oviductal lumen, for the in vivo fertilization of ovulated oocytes. This approach is simpler than the in vitro fertilization-based production of offspring, as the latter always requires an egg transfer to recipient females, which is labor-intensive and time-consuming. In this review, we summarize the various methods for AI reported so far, the history of sperm-mediated gene transfer, a technology to produce genetically engineered animals through in vivo fertilization with sperm carrying exogenous DNA, and finally describe the possibility of AI-mediated creation of GE animals using GE sperm. <a href="/2673-6284/13/4/45">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Section <a href="/journal/biotech/sections/biotechnology_regulation">Biotechnology Regulation</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/45/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1518151"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1518151"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1518151" data-cycle-prev="#prev1518151" data-cycle-progressive="#images1518151" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1518151-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00045/article_deploy/html/images/biotech-13-00045-ag-550.jpg?1731380122" alt="" style="border: 0;"><p>Graphical abstract</p></div><script id="images1518151" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1518151-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00045/article_deploy/html/images/biotech-13-00045-g001-550.jpg?1731380116'><p>Figure 1</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1518151-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00045/article_deploy/html/images/biotech-13-00045-g002-550.jpg?1731380118'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1518151-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00045/article_deploy/html/images/biotech-13-00045-g003-550.jpg?1731380119'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1518151-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00045/article_deploy/html/images/biotech-13-00045-g004-550.jpg?1731380121'><p>Figure 4</p></div></script></div></div><div id="article-1518151-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00045/article_deploy/html/images/biotech-13-00045-ag-550.jpg?1731380122" title=" <strong>Graphical abstract</strong><br/><strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/45'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00045/article_deploy/html/images/biotech-13-00045-g001-550.jpg?1731380116" title=" <strong>Figure 1</strong><br/> <p>Schematic representation of sperm-mediated gene transfer–egg transfer (SMGT-ET). To obtain sperms transfected with exogenous nucleic acids, epididymal sperms were first isolated in a drop (in vitro fertilization (IVF) medium) covered with paraffin oil. Part of these sperms was then subjected to brief incubation with nucleic acids (i.e., plasmid DNA) and gene delivery-enhancing reagents (such as DMSO, liposomes, and nanoparticles). These transfected sperms were then subjected to IVF to obtain fertilized eggs, which were then cultivated up to two-cell embryos. These two-cell embryos are then transferred to the oviduct of a pseudopregnant recipient female for further development, ET.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/45'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00045/article_deploy/html/images/biotech-13-00045-g002-550.jpg?1731380118" title=" <strong>Figure 2</strong><br/> <p>Various methods of artificial insemination (AI). (<b>A</b>) Schematic representation of surgical AI based on intraoviductal transfer of sperm, IOTS, and intrabursal transfer of sperm, ITS (<b>a</b>). Both techniques were performed under a dissecting microscope (<b>b</b>), using a mouth-controlled micropipette (<b>c</b>). This illustration is based on studies by Sato et al. [<a href="#B69-biotech-13-00045" class="html-bibr">69</a>] and Sato and Kimura [<a href="#B70-biotech-13-00045" class="html-bibr">70</a>]. (<b>B</b>) Schematic representation of non-surgical AI using the non-surgical embryo and sperm transfer (mNSET) device in unanesthetized females. First, the small speculum tip (<b>a</b>) was inserted into the vagina of the female, as shown in (<b>b</b>). Second, the tip containing 40 μL of sperm was inserted into the small speculum using Pipetman (<b>c</b>) for releasing sperm within a uterine horn (<b>d</b>). This illustration is based on Stone et al. [<a href="#B66-biotech-13-00045" class="html-bibr">66</a>] and Stone [<a href="#B72-biotech-13-00045" class="html-bibr">72</a>].</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/45'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00045/article_deploy/html/images/biotech-13-00045-g003-550.jpg?1731380119" title=" <strong>Figure 3</strong><br/> <p>Time schedule for intrabursal transfer of sperm (ITS) (<b>A</b>) and non-surgical AI (<b>B</b>). The surgical AI (ITS) was conducted as previously established [<a href="#B71-biotech-13-00045" class="html-bibr">71</a>]. A low concentration of PMSG (2–0.2 IU) was administered at 9 AM to induce ovulation, similar to natural ovulation conditions (indicated by asterisks). The time schedule for non-surgical AI was based on Stone et al. [<a href="#B66-biotech-13-00045" class="html-bibr">66</a>].</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/45'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00045/article_deploy/html/images/biotech-13-00045-g004-550.jpg?1731380121" title=" <strong>Figure 4</strong><br/> <p>Schematic of sperm-mediated gene transfer–artificial insemination (SMGT-AI)-based genome editing (SMGT-AI-GE). To obtain sperms transfected with exogenous nucleic acids, isolated epididymal sperms were first subjected to brief incubation in a solution containing CRISPR/Cas9 components, gene delivery-enhancing reagents (such as DMSO, liposomes, and microparticles), and fluorescent marker expression plasmid DNA for a short period. The solution containing the transfected sperm was then subjected to AI (ITS) in superovulated females, 7 h after hCG administration. A low concentration of PMSG (2–0.2 IU) was administered at 9 AM to induce ovulation, similar to natural ovulation conditions (indicated by asterisks). Cleavage-stage embryos were collected to examine the presence/expression of the transgene (plasmid) and possible mutations in the target locus. In such cases, AI-treated females are permitted to deliver their pups. Genotyping of these pups may reveal genome editing at the target locus in F0 pups.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/45'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1506550" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 16 pages, 3361 KiB   </span> <a href="/2673-6284/13/4/44/pdf?version=1729851043" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Ultraviolet, Did the Cell See It from the Side or the Bottom? Assessment and Modeling of UV Effects on Cultured Cells Using the CL-1000 UV-Crosslinker" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <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> <a class="title-link" href="/2673-6284/13/4/44">Ultraviolet, Did the Cell See It from the Side or the Bottom? Assessment and Modeling of UV Effects on Cultured Cells Using the CL-1000 UV-Crosslinker</a> <div class="authors"> by <span class="inlineblock "><strong>Takahiro Oyama</strong>, </span><span class="inlineblock "><strong>Kai Yanagihara</strong>, </span><span class="inlineblock "><strong>Anna Arai</strong>, </span><span class="inlineblock "><strong>Takanori Kamiya</strong>, </span><span class="inlineblock "><strong>Midori Oyama</strong>, </span><span class="inlineblock "><strong>Takashi Tanikawa</strong>, </span><span class="inlineblock "><strong>Takehiko Abe</strong> and </span><span class="inlineblock "><strong>Tomomi Hatanaka</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 44; <a href="https://doi.org/10.3390/biotech13040044">https://doi.org/10.3390/biotech13040044</a> - 25 Oct 2024 </div> Viewed by 772 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Numerous natural extracts and compounds have been evaluated for their ability to mitigate the adverse effects of ultraviolet (UV) overexposure. However, variability in the UV doses that trigger biological responses across studies likely arises from inconsistencies in UV exposure standardization. We hypothesize that <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/44/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Numerous natural extracts and compounds have been evaluated for their ability to mitigate the adverse effects of ultraviolet (UV) overexposure. However, variability in the UV doses that trigger biological responses across studies likely arises from inconsistencies in UV exposure standardization. We hypothesize that these discrepancies are due to variations in culture plates and dishes. The UV dose (D) required to reduce cell viability by 50% differed by a factor of ten between 3.5 cm dishes and 96-well plates. Similarly, the EC<sub>50</sub> dose for IL-6 release (<i>D</i><sub>1/2</sub>) varied, potentially correlating with the surface area (S). UV exposure to wells with increasing height in 3.5 cm dishes resulted in a decrease in IL-6 release, suggesting that the greater the well height, the more it may influence UV exposure through reflection or shielding effects, thereby contributing to the physiological effects on the cells. To compare these differences among plates, we defined the height-to-diameter ratio (r). Analysis revealed a linear correlation between <i>D</i><sub>1/2</sub> and S in a log-log plot, and between <i>D</i><sub>1/2</sub> and r in a semi-log plot. From this, we defined two empirical indices <i>σ</i> and ρ for UV dose adjustment. A deductive model was also developed to derive a D′ value that adjusts UV doses without requiring training. As with σ and ρ, the UV dose D was effectively adjusted using D′ as well. These attempts suggest that D′ offers a foundational framework for evaluating UVB effects on cultured cells. <a href="/2673-6284/13/4/44">Full article</a> </div> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/44/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1506550"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1506550"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1506550" data-cycle-prev="#prev1506550" data-cycle-progressive="#images1506550" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1506550-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g001-550.jpg?1729851110" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1506550" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1506550-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g002-550.jpg?1729851112'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1506550-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g003-550.jpg?1729851113'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1506550-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g004-550.jpg?1729851116'><p>Figure 4</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1506550-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g005-550.jpg?1729851116'><p>Figure 5</p></div> --- <div class='openpopupgallery' data-imgindex='5' data-target='article-1506550-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g006-550.jpg?1729851117'><p>Figure 6</p></div> --- <div class='openpopupgallery' data-imgindex='6' data-target='article-1506550-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g007-550.jpg?1729851119'><p>Figure 7</p></div> --- <div class='openpopupgallery' data-imgindex='7' data-target='article-1506550-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g0A1-550.jpg?1729851121'><p>Figure A1</p></div></script></div></div><div id="article-1506550-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g001-550.jpg?1729851110" title=" <strong>Figure 1</strong><br/> <p>Specifications of the CL-1000 UV Crosslinker. The dimensions of this equipment are width = 25.4 cm, depth = 30.5 cm, and height = 12.7 cm. The equipment accommodates up to five fluorescent tubes with UVB emission at 302 nm. The dotted line indicates a UV lamp that was detached in this study. The inner walls of the equipment can reflect UV radiation, allowing plates to receive UVB both directly and indirectly. A detector located at the inner back right side of the equipment monitors the exposure automatically once the UV dose reaches the preset value.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/44'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g002-550.jpg?1729851112" title=" <strong>Figure 2</strong><br/> <p>The dose-dependent cytotoxic effects of UVB on cultured keratinocytes. NHEK cells were seeded in both 3.5 cm dishes and 96-well black plates as indicated in <a href="#biotech-13-00044-t002" class="html-table">Table 2</a>. After a 2-day preculture, the indicated doses of UVB were irradiated to the wells. Cells were subjected to each experiment after an additional 1-day incubation. (<b>a</b>,<b>b</b>) Cells in 3.5 cm dishes were detached and counted under a microscope, mixed with an equal volume of 0.4% <span class="html-italic">v</span>/<span class="html-italic">v</span> trypan blue dye. The relative numbers of (<b>a</b>) total cells and (<b>b</b>) trypan blue-negative cell rates (%) were calculated. (<b>c</b>,<b>d</b>) Cells in 96-well plates were (<b>c</b>) stained with Hoechst 33342 for 30 min or (<b>d</b>) incubated with WST-8 reagent for 60 min. Individual quantified values are represented as gray dots on the bar chart. All data are expressed as the mean ± SD of at least three independent experiments. Statistical analyses were performed using Dunnett’s test, compared to the 0 mJ/cm<sup>2</sup> group. * <span class="html-italic">p</span> &lt; 0.05.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/44'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g003-550.jpg?1729851113" title=" <strong>Figure 3</strong><br/> <p>The dose-dependent inflammatory effects of UVB on cultured keratinocytes. NHEK cells were seeded in (<b>a</b>) 3.5 cm dishes, (<b>b</b>) 12-well, (<b>c</b>) 24-well, and (<b>d</b>) 96-well plates as indicated in <a href="#biotech-13-00044-t002" class="html-table">Table 2</a>. After a 2-day preculture, the indicated doses of UVB were irradiated to the wells. Following a 24 h incubation, IL-6 concentrations in the supernatant medium were measured by ELISA. All data are expressed as the mean ± SD of at least three independent experiments (blue dots). Data excluding the highest dose group were fitted with a 4-parameter sigmoid curve, indicated by the red line. The parameters predicted by the calculation are shown in <a href="#biotech-13-00044-t003" class="html-table">Table 3</a>.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/44'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g004-550.jpg?1729851116" title=" <strong>Figure 4</strong><br/> <p>Investigation of the change in IL-6 release based on the solid angle and height of the well wall. (<b>a</b>) Cross-sectional view from the side of the CL-1000. A well placed in the center of the equipment is schematically depicted. The height and radius of the well are expressed as h and a cm, respectively. (<b>b</b>) The conceptual drawing of the calculation of the solid angle in the case of stacking dishes. (<b>c</b>) Pictures of stacking dishes. (<b>d</b>) The 5.0 × 10<sup>4</sup> NHEK cells were seeded in 3.5 cm dishes under the conditions indicated in (<b>c</b>). After a 2-day preculture, 10 mJ/cm<sup>2</sup> of UVB was irradiated to the wells. Following a 24 h incubation, IL-6 concentrations in the supernatant medium were measured by ELISA. (<b>e</b>) Pictures of covering dishes with a cylinder made of black paper. (<b>f</b>) The 5.0 × 10<sup>4</sup> NHEK cells were seeded in 3.5 cm dishes under the conditions indicated in (<b>e</b>). After a 2-day preculture, 20 mJ/cm<sup>2</sup> of UVB was irradiated to the wells. Following a 24 h incubation, IL-6 concentrations in the supernatant medium were measured by ELISA. Individual quantified values are represented as gray dots on the bar chart. All data were expressed as the mean ± SD of at least three independent experiments. Statistical analyses were performed using Dunnett’s test, compared to the 1.3 cm-covered group (<b>g</b>). * <span class="html-italic">p</span> &lt; 0.05.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/44'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g005-550.jpg?1729851116" title=" <strong>Figure 5</strong><br/> <p>Correlation between S and <span class="html-italic">D</span><sub>1/2</sub>, and between r and <span class="html-italic">D</span><sub>1/2</sub>. (<b>a</b>) Scatter plot between log<sub>10</sub>(S) and log<sub>10</sub>(<span class="html-italic">D</span><sub>1/2</sub>) with a regression line described by the formula y = −0.6869x + 1.8634. (<b>b</b>) The scatter plot between r and log<sub>10</sub>(<span class="html-italic">D</span><sub>1/2</sub>) with a regression line described by the formula y = 0.6988x + 0.9329.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/44'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g006-550.jpg?1729851117" title=" <strong>Figure 6</strong><br/> <p>Absorbance change with the serial dilutions of HuMedia-KG2. The medium specialized for keratinocytes (HuMedia-KG2) was serially diluted and absorbance measured at 302 nm. Since the molar concentration was not defined, the relative concentration of the undiluted solution was taken as 1. The data were fitted with a linear regression, setting the intercept to 0.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/44'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g007-550.jpg?1729851119" title=" <strong>Figure 7</strong><br/> <p>Comparison of the corrected dose–effect plot using three indices as dose. (<b>a</b>) IL-6 release by UVB exposure from <a href="#biotech-13-00044-f004" class="html-fig">Figure 4</a> re-plotted in one figure. (<b>b</b>–<b>d</b>) IL-6 response curves using (<b>b</b>) <span class="html-italic">σ</span>, (<b>c</b>) ρ, and (<b>d</b>) D′ as the <span class="html-italic">X</span>-axis. Data for 3.5 cm dish, 12-well, 24-well, and 96-well plates are represented as mustard circles, gray squares, orange triangles, and blue diamonds, respectively. Each dataset was fitted with a 4-parameter sigmoid curve shown by the smooth lines with same colors. All data are expressed as mean ± SD. Data from all plates were combined and fitted with a 4-parameter sigmoid curve, depicted by the black line. The R<sup>2</sup> values of the fittings for all values are shown in the upper left of each graph.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/44'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00044/article_deploy/html/images/biotech-13-00044-g0A1-550.jpg?1729851121" title=" <strong>Figure A1</strong><br/> <p>Schematic diagram of wells on culture plates and dishes. A well is placed in the <math display="inline"><semantics> <mrow> <mi>ρ</mi> <mi>ψ</mi> <mi>z</mi> </mrow> </semantics></math> -space with the center of its bottom surface at origin O. (<b>a</b>) Spherical coordinates placed in a well. As all UVB rays pass through the corresponding surface of the hemisphere, the dose at point X is calculated by integrating over spherical coordinates. The z’-axis is parallel to <span class="html-italic">z</span>-axis passing through X. Left panel shows an illustration of area element calculation. The area element <math display="inline"><semantics> <mrow> <mi mathvariant="normal">d</mi> <mi mathvariant="sans-serif">Ω</mi> <mo>=</mo> <mrow> <mrow> <mi mathvariant="normal">sin</mi> </mrow> <mo>⁡</mo> <mrow> <mi>θ</mi> </mrow> </mrow> <mi>d</mi> <mi mathvariant="sans-serif">θ</mi> <mi>d</mi> <mi>ϕ</mi> </mrow> </semantics></math>. (<b>b</b>) A cross-section of a well cut by a plane passing through a specific point X and inclined at an angle φ. An intersection point of the circle O and the plane directed to <math display="inline"><semantics> <mrow> <mi>ρ</mi> <mo>&gt;</mo> <mn>0</mn> <mo> </mo> </mrow> </semantics></math>is named Y. s is the length of XY. (<b>c</b>) A top view of the well. (<b>d</b>) Cross-section of a well cut by the <math display="inline"><semantics> <mrow> <mi>ρ</mi> <mi>z</mi> </mrow> </semantics></math>-plane. a, radius of the well; h, height of the well; l, height of the filled medium.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/44'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1502253" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 13 pages, 1793 KiB   </span> <a href="/2673-6284/13/4/43/pdf?version=1729325807" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Microbial Protein and Metabolite Profiles of Klebsiella oxytoca M5A1 in a Bubble Column Bioreactor" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <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> <a class="title-link" href="/2673-6284/13/4/43">Microbial Protein and Metabolite Profiles of <i>Klebsiella oxytoca M5A1</i> in a Bubble Column Bioreactor</a> <div class="authors"> by <span class="inlineblock "><strong>Tawakalt Ayodele</strong>, </span><span class="inlineblock "><strong>Musiliu Liadi</strong>, </span><span class="inlineblock "><strong>Abodunrin Tirmidhi Tijani</strong>, </span><span class="inlineblock "><strong>Kudirat Alarape</strong>, </span><span class="inlineblock "><strong>Christiana Bitrus</strong>, </span><span class="inlineblock "><strong>Clairmont L. Clementson</strong> and </span><span class="inlineblock "><strong>Ademola Hammed</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 43; <a href="https://doi.org/10.3390/biotech13040043">https://doi.org/10.3390/biotech13040043</a> - 19 Oct 2024 </div> Viewed by 625 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> The production of microbial proteins (MPs) has emerged as a critical focus in biotechnology, driven by the need for sustainable and scalable alternatives to traditional protein sources. This study investigates the efficacy of two experimental setups in producing MPs using the nitrogen-fixing bacterium <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/43/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> The production of microbial proteins (MPs) has emerged as a critical focus in biotechnology, driven by the need for sustainable and scalable alternatives to traditional protein sources. This study investigates the efficacy of two experimental setups in producing MPs using the nitrogen-fixing bacterium <i>Klebsiella oxytoca M5A1</i>. <i>K. oxytoca M5A1</i>, known for its facultative anaerobic growth and capability to fix atmospheric nitrogen, offers a promising avenue for environmentally friendly protein production. This research compares the performance of a simple bubble column (BC) bioreactor, which promotes efficient mixing and cross-membrane gas transfer, with static fermentation, a traditional method lacking agitation and aeration. The study involved the parallel cultivation of <i>K. oxytoca M5A1</i> in both systems, with key parameters such as microbial growth, glucose utilization, protein concentration, and metabolite profiles monitored over a 48 h period. The results indicate that the BC bioreactor consistently outperformed static fermentation regarding the growth rate, protein yield, and glucose utilization efficiency. The BC exhibited a significant increase in protein production, reaching 299.90 µg/mL at 48 h, compared to 219.44 µg/mL in static fermentation. The organic acid profile reveals both synthesis and utilization regimes of varying patterns. These findings highlight the advantages of the BC bioreactor for MP production, particularly its ability to maintain aerobic conditions that support higher growth and yield. <a href="/2673-6284/13/4/43">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Section <a href="/journal/biotech/sections/industrial_biotechnology">Industrial Biotechnology</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/43/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1502253"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1502253"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1502253" data-cycle-prev="#prev1502253" data-cycle-progressive="#images1502253" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1502253-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00043/article_deploy/html/images/biotech-13-00043-g001-550.jpg?1729325987" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1502253" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1502253-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00043/article_deploy/html/images/biotech-13-00043-g002-550.jpg?1729325990'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1502253-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00043/article_deploy/html/images/biotech-13-00043-g003-550.jpg?1729325992'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1502253-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00043/article_deploy/html/images/biotech-13-00043-g004-550.jpg?1729325993'><p>Figure 4</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1502253-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00043/article_deploy/html/images/biotech-13-00043-g005-550.jpg?1729325996'><p>Figure 5</p></div></script></div></div><div id="article-1502253-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00043/article_deploy/html/images/biotech-13-00043-g001-550.jpg?1729325987" title=" <strong>Figure 1</strong><br/> <p>Fabricated bubble column bioreactor.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/43'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00043/article_deploy/html/images/biotech-13-00043-g002-550.jpg?1729325990" title=" <strong>Figure 2</strong><br/> <p>Growth profile of <span class="html-italic">K. oxytoca M5A1</span> under BC and static conditions. The data are presented with absorbance mean values ± SD of n = 3 experiments.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/43'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00043/article_deploy/html/images/biotech-13-00043-g003-550.jpg?1729325992" title=" <strong>Figure 3</strong><br/> <p>Glucose utilization pattern of <span class="html-italic">K. oxytoca M5A1</span> under the BC and static conditions. The data are presented with absorbance mean values ± SD of n = 3 experiments.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/43'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00043/article_deploy/html/images/biotech-13-00043-g004-550.jpg?1729325993" title=" <strong>Figure 4</strong><br/> <p>Microbial protein concentration in the BC and static conditions. The data are presented with mean values ± SD of n = 3 experiments.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/43'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00043/article_deploy/html/images/biotech-13-00043-g005-550.jpg?1729325996" title=" <strong>Figure 5</strong><br/> <p>Organic acid production in the BC reactor and static fermentation. The data are presented with mean values ± SD of n = 3 experiments.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/43'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1496445" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 18 pages, 3640 KiB   </span> <a href="/2673-6284/13/4/42/pdf?version=1728639397" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Phytochemical Composition, Antioxidant, Anti-Helicobacter pylori, and Enzyme Inhibitory Evaluations of Cleistocalyx operculatus Flower Bud and Leaf Fractions" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <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> <a class="title-link" href="/2673-6284/13/4/42">Phytochemical Composition, Antioxidant, Anti-<i>Helicobacter pylori</i>, and Enzyme Inhibitory Evaluations of <i>Cleistocalyx operculatus</i> Flower Bud and Leaf Fractions</a> <div class="authors"> by <span class="inlineblock "><strong>Doan Thien Thanh</strong>, </span><span class="inlineblock "><strong>Mai Thanh Tan</strong>, </span><span class="inlineblock "><strong>Nguyen Thi My Thu</strong>, </span><span class="inlineblock "><strong>Pham Nhat Phuong Trinh</strong>, </span><span class="inlineblock "><strong>Pham Thi Hoai Thuong</strong>, </span><span class="inlineblock "><strong>Pham Thi Giang Tuyet</strong>, </span><span class="inlineblock "><strong>Luong Thi My Ngan</strong> and </span><span class="inlineblock "><strong>Tran Trung Hieu</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 42; <a href="https://doi.org/10.3390/biotech13040042">https://doi.org/10.3390/biotech13040042</a> - 11 Oct 2024 </div> Viewed by 889 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Six solvent fractions isolated from flower bud and leaf ethanolic extracts of <i>Cleistocalyx operculatus</i> were analyzed for their phytochemical contents, including phenolics, flavonoids, saponins, tannins, and alkaloids. Antioxidant activities were measured using the ABTS, DPPH, and FRAP assays. The results showed that the <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/42/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Six solvent fractions isolated from flower bud and leaf ethanolic extracts of <i>Cleistocalyx operculatus</i> were analyzed for their phytochemical contents, including phenolics, flavonoids, saponins, tannins, and alkaloids. Antioxidant activities were measured using the ABTS, DPPH, and FRAP assays. The results showed that the flower bud aqueous fraction (BAF) and the leaf aqueous fraction (LAF) rich in phenolic content (768.18 and 490.74 mg GAE/g dry extract, respectively) exhibited significantly higher antioxidant activities than the other fractions. The flower bud hexane fraction (BHF) had remarkably high flavonoid and saponin contents (134.77 mg QE/g and 153.33 mg OA/g dry extract, respectively), followed by that of the leaf hexane fraction (LHF) (76.54 mg QE/g and 88.25 mg OA/g dry extract, respectively). The BHF and LHF were found to have extremely high antibacterial activity against two <i>H. pylori</i> strains, ATCC 51932 and 43504 (MICs of 125 µg/mL). Interestingly, DMC (2′,4′-Dihydroxy-6′-methoxy-3′,5′-dimethylchalcone) isolated from the BHF displayed greater antibacterial activity against the bacterial strains (MICs of 25–50 µg/mL) than those of the fractions. In addition, DMC presented potent inhibitory effects on <i>H. pylori</i> urease (IC<sub>50</sub> of 3.2 µg/mL) and α-amylase (IC<sub>50</sub> of 83.80 µg/mL), but no inhibition against α-glucosidase. It was also demonstrated that DMC showed pronounced inhibitory effects on the urease activity and biofilm formation of <i>H. pylori</i>, and could increase the membrane permeability of the bacterial cells. Scanning electron micrographs depicted that the BHF and DMC had strong effects on the cell shape and significantly induced the distortion and damage of the cell membrane. The fractions and DMC showed no significant toxicity to four tested human cell lines. Efforts to reduce antibiotic use indicate the need for further studies of the flower buds and DMC as potential products to prevent or treat gastric <i>H. pylori</i> infections. <a href="/2673-6284/13/4/42">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Special Issue <a href=" /journal/biotech/special_issues/7KP0712M2L ">Natural Antioxidants: Determination in Food and Nutraceuticals and Implications on Human Health</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/42/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1496445"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1496445"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1496445" data-cycle-prev="#prev1496445" data-cycle-progressive="#images1496445" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1496445-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00042/article_deploy/html/images/biotech-13-00042-ag-550.jpg?1728639523" alt="" style="border: 0;"><p>Graphical abstract</p></div><script id="images1496445" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1496445-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00042/article_deploy/html/images/biotech-13-00042-g001-550.jpg?1728639515'><p>Figure 1</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1496445-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00042/article_deploy/html/images/biotech-13-00042-g002-550.jpg?1728639521'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1496445-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00042/article_deploy/html/images/biotech-13-00042-g003-550.jpg?1728639523'><p>Figure 3</p></div></script></div></div><div id="article-1496445-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00042/article_deploy/html/images/biotech-13-00042-ag-550.jpg?1728639523" title=" <strong>Graphical abstract</strong><br/><strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/42'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00042/article_deploy/html/images/biotech-13-00042-g001-550.jpg?1728639515" title=" <strong>Figure 1</strong><br/> <p>Effect of <span class="html-italic">C. operculatus</span> fractions and DMC at sub-MICs on <span class="html-italic">H. pylori</span> biofilm formation 48 h post-treatment. Data are reported as means ± SD.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/42'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00042/article_deploy/html/images/biotech-13-00042-g002-550.jpg?1728639521" title=" <strong>Figure 2</strong><br/> <p>SEM micrographs of <span class="html-italic">H. pylori</span> ATCC 43504 depicting untreated cells (<b>A</b>,<b>B</b>) and cells treated with 125 µg/mL of BHF (<b>C</b>,<b>D</b>) and 50 µg/mL of DMC (<b>E</b>,<b>F</b>); bars 5 and 1 µm, respectively.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/42'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00042/article_deploy/html/images/biotech-13-00042-g003-550.jpg?1728639523" title=" <strong>Figure 3</strong><br/> <p>Effect of <span class="html-italic">C. operculatus</span> fractions and DMC at sub-MICs on the uptake of crystal violet by <span class="html-italic">H. pylori</span> ATCC 43504 after 2 h of treatment. Data are reported as means ± SD (n = 3).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/42'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1491873" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 16 pages, 2581 KiB   </span> <a href="/2673-6284/13/4/41/pdf?version=1729148667" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Loss of Cell-Cell Contact Inhibits Cellular Differentiation of α-Catenin Knock Out P19 Embryonal Carcinoma Cells and Their Colonization into the Developing Mouse Embryos" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <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> <a class="title-link" href="/2673-6284/13/4/41">Loss of Cell-Cell Contact Inhibits Cellular Differentiation of α-Catenin Knock Out P19 Embryonal Carcinoma Cells and Their Colonization into the Developing Mouse Embryos</a> <div class="authors"> by <span class="inlineblock "><strong>Masahiro Sato</strong>, </span><span class="inlineblock "><strong>Emi Inada</strong>, </span><span class="inlineblock "><strong>Naoko Kubota</strong> and </span><span class="inlineblock "><strong>Masayuki Ozawa</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 41; <a href="https://doi.org/10.3390/biotech13040041">https://doi.org/10.3390/biotech13040041</a> - 3 Oct 2024 </div> Viewed by 1009 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Cadherin−catenin cell−cell adhesion complexes, composed of cadherin, β-catenin or plakoglobin, and α-catenin (α-cat) molecules, are crucial for maintaining cell−cell contact and are commonly referred to as “adherens junctions (AJs).” Inactivating this system leads to loss of cell−cell contact and developmental arrest in early <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/41/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Cadherin−catenin cell−cell adhesion complexes, composed of cadherin, β-catenin or plakoglobin, and α-catenin (α-cat) molecules, are crucial for maintaining cell−cell contact and are commonly referred to as “adherens junctions (AJs).” Inactivating this system leads to loss of cell−cell contact and developmental arrest in early embryos. However, it remains unclear whether the loss of cell−cell contact affects the differentiation of embryonic cells. In this study, we explored the use of a murine embryonal carcinoma cell line, P19, as an in vitro model for early embryogenesis. P19 cells easily form embryoid bodies (EBs) and are susceptible to cellular differentiation in response to retinoic acid (RA) and teratoma formation. Using CRISPR/Cas9 technology to disrupt the endogenous <i>α-cat</i> gene in P19 cells, we generated <i>α-cat</i> knockout (KO) cells that exhibited a loss of cell−cell contact. When cultivated on non-coated dishes, these <i>α-cat</i> KO cells formed EBs, but their structures were labile. In the RA-containing medium, the <i>α-cat</i> KO EBs failed to produce differentiated cells on their outer layer and continued to express SSEA-1, an antigen specific to pluripotent cells. Teratoma formation assays revealed an absence of overt differentiated cells in tumors derived from <i>α-cat</i> KO P19 cells. Aggregation assays revealed the inability of the KO cells to colonize into the zona pellucida-denuded 8-cell embryos. These findings suggest that the AJs are essential for promoting the early stages of cellular differentiation and for the colonization of early-developing embryos. <a href="/2673-6284/13/4/41">Full article</a> </div> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/41/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1491873"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1491873"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1491873" data-cycle-prev="#prev1491873" data-cycle-progressive="#images1491873" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1491873-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-ag-550.jpg?1729148752" alt="" style="border: 0;"><p>Graphical abstract</p></div><script id="images1491873" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1491873-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g001-550.jpg?1729148744'><p>Figure 1</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1491873-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g002-550.jpg?1729148747'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1491873-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g003-550.jpg?1729148748'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1491873-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g004-550.jpg?1729148748'><p>Figure 4</p></div> --- <div class='openpopupgallery' data-imgindex='5' data-target='article-1491873-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g005-550.jpg?1729148751'><p>Figure 5</p></div> --- <div class='openpopupgallery' data-imgindex='6' data-target='article-1491873-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g006-550.jpg?1729148751'><p>Figure 6</p></div></script></div></div><div id="article-1491873-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-ag-550.jpg?1729148752" title=" <strong>Graphical abstract</strong><br/><strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/41'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g001-550.jpg?1729148744" title=" <strong>Figure 1</strong><br/> <p>Molecules required for cell–cell contact and plasmids for the generation of genetically engineered P19 cells. (<b>A</b>): Multiprotein complexes at adherens junctions (AJs). α-Catenin (α) is one of the essential components in the E-cadherin–catenin cell adhesion complex, which mediates cell–cell contact via the formation of AJs. It forms a bridge between the E-cadherin and cytoskeletal actin filaments. Once α is depleted from a cell, the bridge would be disrupted together with loss of mechanotension, leading to loss of cell–cell contact. β, β-catenin. (<b>B</b>): The vectors for yielding <span class="html-italic">α-cat</span> knock out (KO) P19 cells using the CRISPR/Cas9 system or those for rescuing <span class="html-italic">α-cat</span> KO cells. Structure of pCGSap1 carrying a <span class="html-italic">Cas9</span> expression unit (comprising <span class="html-italic">CAG</span>, the second intron of the rabbit <span class="html-italic">β-globin</span> gene, the humanized <span class="html-italic">Cas9</span> (<span class="html-italic">hCas9</span>) gene, and the poly(A) site of the rabbit <span class="html-italic">β-globin</span> gene) and a guide RNA (gRNA) expression unit (comprising <span class="html-italic">U6</span>, multiple sites into which chemically synthesized gRNA is inserted, and a poly(A) site). Oligonucleotides targeting <span class="html-italic">α-cat</span> exon 1 were synthesized and inserted into the <span class="html-italic">Sap</span> I site of pCGSap1 to create pCGSap1-<span class="html-italic">α-cat</span>. The underlined sequence corresponds to the sequence of gRNA targeting the <span class="html-italic">α-cat</span> gene. Nucleotides (shown in red and green) at both the 5′ and 3′ ends of the gRNA are those recognized by <span class="html-italic">Sap</span> I. pC-bsr is a vector conferring expression of the blasticidin S resistance gene (<span class="html-italic">bsr</span>). pC-GFPHA is a vector that confers the expression of enhanced green fluorescent protein (EGFP) and an HA tag (HA). pC-ha-cat is a vector conferring expression of human α-cat cDNA and HA. CAG, chicken β-actin based promoter; <span class="html-italic">U6</span>, human <span class="html-italic">U6</span> promoter. <span class="html-italic">neo</span>, neomycin resistance gene expression unit; HA, HA tag; pA, poly(A) site.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/41'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g002-550.jpg?1729148747" title=" <strong>Figure 2</strong><br/> <p>Morphology of the genetically engineered P19 cells, western blotting, and RNA expression profiling. (<b>A</b>): Cell behavior of P19 cells (a,d) and <span class="html-italic">α-catenin</span> (<span class="html-italic">α-cat</span>) KO P19 cells [C7 (b,e) and C9 (c,f)] 2 days after seeding onto an adhesive substrate densely (a–c) or sparsely (d–f). Bar: 100 μm. (<b>B</b>): Immunoblot detection of α-cat in P19, C7, and revertant (rev-C7) cells. α-Tubulin was used as a loading control. (<b>C</b>): Cell behavior of P19 cells (a) and <span class="html-italic">α-cat</span> KO P19 cells (C7) (b) and revertant (rev-C7) (c) 4 days after seeding onto an adhesive substrate densely. Bar: 100 μm. (<b>D</b>): Heatmap analysis of P19, C7, and rev-C7 cells.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/41'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g003-550.jpg?1729148748" title=" <strong>Figure 3</strong><br/> <p>Embryoid body (EB) formation of P19 cells. (<b>A</b>): EB formation by simple suspension culture of P19 cells (a–c) and C7 cells (d–f) in a non-coated dish for 2 (a,d), 4 (b,e), and 8 d (c,f). Over 10 EBs were checked and they all showed similar morphology. Bar: 100 μm. (<b>B</b>): Immunostaining with an anti-SSEA-1 antibody. EBs were generated from P19 cells (a,b) or C7 cells (c,d) after cultivation in DMEM-high glucose/10% FBS in a non-adhesive dish for 8 days. Arrows and asterisks indicate positive staining with anti-SSEA-1. (a,c): Photographs captured under light; (b,d): Photographs captured under UV illumination. Bar: 100 μm.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/41'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g004-550.jpg?1729148748" title=" <strong>Figure 4</strong><br/> <p>Retinoic acid (RA)-induced differentiation of P19 embryoid bodies (EBs). (<b>A</b>): EBs generated from P19 cells or C7 after cultivation in a non-adhesive dish in the presence of RA at various concentrations (10<sup>−8</sup> or 10<sup>−5</sup> M) for 4 days (a–d), 6 days (e–h), and 9 days (i–k). Bar: 100 μm. (<b>B</b>): Immunostaining with an anti-SSEA-1 antibody. EBs generated from P19 cells (a,b,e,f) or C7 cells (c,d) after cultivation in the presence of 10<sup>−8</sup> M (a–d) or 10<sup>−5</sup> M RA (e,f) in a non-adhesive dish for 8 days. The outer layer surrounding the core cells (indicated by asterisks) of P19 EBs treated with 10<sup>−8</sup> M or 10<sup>−5</sup> M RA was completely negative for anti-SSEA-1 staining (a,b,e,f). In contrast, C7 EBs treated with 10<sup>−8</sup> M RA were reactive to anti-SSEA-1 in both the outer layer (arrows in d) and the inner core (asterisks in d). Bar: 100 μm. (<b>C</b>): Outgrowth of P19 cells (a,b) and C7 cells (c) four days after attachment of EBs [grown for eight days in the presence of 10<sup>−8</sup> M (a,c) or 10<sup>−5</sup> M RA (b)] to the adhesive surface. Various types of differentiated cells [elongated cells (a’) vs. flat cells (b’)] were detected when P19 cells were cultured with different concentrations of RA. In contrast, no differentiated cells (c’) were detected when C7 cells were cultivated with 10<sup>−8</sup> M RA. Bar: 100 μm. (<b>D</b>): Quantitative analysis of cells at the edge of a colony. Cells in the boxes (a’–c’) presented in <a href="#biotech-13-00041-f004" class="html-fig">Figure 4</a>C were subjected to image analysis and plotted. Over 6 (for P19 cells treated with 10<sup>−8</sup> M RA) and 10 cells (for P19 cells treated with 10<sup>−5</sup> M RA and C7 treated with 10<sup>−8</sup> M RA) were selected randomly and plotted. **: <span class="html-italic">p</span> &lt; 0.05.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/41'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g005-550.jpg?1729148751" title=" <strong>Figure 5</strong><br/> <p>In vivo teratoma formation. (<b>A</b>): Schematic illustration of in vivo teratoma formation by inoculation of tumor cells into the pancreatic parenchyma of nude mice. First, the spleen and pancreas were removed from anesthetized mice. Then, a solution (1~2 μL) containing tumor cells and trypan blue (used as visible marker) is injected into the inner area of pancreas using a breath-controlled micropipette under a dissecting microscope. This procedure is developed by Sato et al. [<a href="#B25-biotech-13-00041" class="html-bibr">25</a>] and named “IPPCT”. (<b>B</b>): Hematoxylin-eosin (H&amp;E)-stained cryostat sections of solid tumors generated 1.5 months after IPPCT with P19 cells (a) or C7 cells (b). Differentiated cells are indicated by dotted lines (numbered #1–#3) together with undifferentiated cells (numbered #4) in the P19 cell-derived tumors (a). In contrast, no distinctly differentiated cells were visible in the C7-derived tumors (b). Bar: 100 μm.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/41'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00041/article_deploy/html/images/biotech-13-00041-g006-550.jpg?1729148751" title=" <strong>Figure 6</strong><br/> <p>Formation of aggregation chimeras between embryonal carcinoma (EC) cells and 8-cell embryos. (<b>A</b>): Location of EC after aggregation of chimeras between P19 cells ((e–h), (e’–h’), (m–p), (m’–p’)) or C7 ((a–d), (a’–d’), (i–l), (i’–l’)) and 8-cell embryos. Since EC cells were labelled with EGFP, which is useful for tracking chimeras, green fluorescence in EC cells was checked at the morula ((a–d), (a’–d’), (e–h), (e’–h’)) and blastocyst ((i–l), (i’–l’), (m–p), (m’–p’)) stages after EC cell/embryo aggregation. Arrows indicate EC attached to the surface of an embryo or incorporated into its internal area. Arrowheads indicate that EC are present near the embryo but are not attached to its surface. (a–d), (e–h), (i–l), (m–p): photographed under UV illumination; (a’–d’), (e’–h’), (i’–l’), (m’–p’): photographed under light. Bar: 100 μm. (<b>B</b>): Graphs showing the percentage of embryos with successful aggregation between EC cells and an embryo per total number of embryos tested. Successful aggregation was determined when embryos with one or more EC on the surface or internal area of the embryo were recognized. P19/8, chimera between P19 cells and 8-cell embryo; C7/8, chimera between C7 and 8-cell embryo. **: <span class="html-italic">p</span> &lt; 0.05.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/41'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1491847" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 16 pages, 2552 KiB   </span> <a href="/2673-6284/13/4/40/pdf?version=1729157120" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Structural Analysis and Substrate Specificity of D-Carbamoylase from Pseudomonas" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <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> <a class="title-link" href="/2673-6284/13/4/40">Structural Analysis and Substrate Specificity of D-Carbamoylase from <i>Pseudomonas</i></a> <div class="authors"> by <span class="inlineblock "><strong>Marina Paronyan</strong>, </span><span class="inlineblock "><strong>Haykanush Koloyan</strong>, </span><span class="inlineblock "><strong>Hovsep Aganyants</strong>, </span><span class="inlineblock "><strong>Artur Hambardzumyan</strong>, </span><span class="inlineblock "><strong>Tigran Soghomonyan</strong>, </span><span class="inlineblock "><strong>Sona Avetisyan</strong>, </span><span class="inlineblock "><strong>Sergey Kocharov</strong>, </span><span class="inlineblock "><strong>Henry Panosyan</strong>, </span><span class="inlineblock "><strong>Vehary Sakanyan</strong> and </span><span class="inlineblock "><strong>Anichka Hovsepyan</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 40; <a href="https://doi.org/10.3390/biotech13040040">https://doi.org/10.3390/biotech13040040</a> - 3 Oct 2024 </div> Viewed by 772 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> The synthesis of enantiomeric forms of D-amino acids can be achieved by a two-step “hydantoinase process” based on the sequential catalysis of substrates by specific enzymes, D-carbamoylase and D-hydantoinase. Here, we describe the structural features of D-carbamoylase from <i>Pseudomonas</i>, the encoded gene <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/40/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> The synthesis of enantiomeric forms of D-amino acids can be achieved by a two-step “hydantoinase process” based on the sequential catalysis of substrates by specific enzymes, D-carbamoylase and D-hydantoinase. Here, we describe the structural features of D-carbamoylase from <i>Pseudomonas</i>, the encoded gene of which was chemically synthesized and cloned into <i>Escherichia coli</i>. A significant fraction of the overexpressed recombinant protein forms insoluble inclusion bodies, which are partially converted to a soluble state upon treatment with N-lauroylsarcosine or upon incubation of cells at 28 °C. Purified His-tagged protein exhibits the highest activity towards N-carbamoyl-D-alanine and N-carbamoyl-D-tryptophan. Comprehensive virtual analysis of the interactions of bulky carbamylated amino acids with D-carbamoylase provided valuable information. Molecular docking analysis revealed the location of the substrate binding site in the three-dimensional structure of D-carbamoylase. Molecular dynamics simulations showed that the binding pocket of the enzyme in complex with N-carbamoyl-D-tryptophan was stabilized within 100 nanoseconds. The free energy data showed that Arg176 and Asn173 formed hydrogen bonds between the enzyme and substrates. The studies of D-carbamoylases and the properties of our previously obtained D-hydantoinase suggest the possibility of developing a harmonized biotechnological process for the production of new drugs and peptide hormones. <a href="/2673-6284/13/4/40">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Topic <a href="/topics/MP2U40PVL4">Microbial Biotechnology Products and Biocatalysis Processes for a Sustainable Bioeconomy</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/40/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1491847"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1491847"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1491847" data-cycle-prev="#prev1491847" data-cycle-progressive="#images1491847" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1491847-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-ag-550.jpg?1729557210" alt="" style="border: 0;"><p>Graphical abstract</p></div><script id="images1491847" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1491847-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g001-550.jpg?1729557072'><p>Figure 1</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1491847-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g002-550.jpg?1729557075'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1491847-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g003-550.jpg?1729557078'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1491847-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g004-550.jpg?1729557080'><p>Figure 4</p></div> --- <div class='openpopupgallery' data-imgindex='5' data-target='article-1491847-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g005-550.jpg?1729557082'><p>Figure 5</p></div> --- <div class='openpopupgallery' data-imgindex='6' data-target='article-1491847-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g006-550.jpg?1729557084'><p>Figure 6</p></div></script></div></div><div id="article-1491847-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-ag-550.jpg?1729557210" title=" <strong>Graphical abstract</strong><br/><strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/40'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g001-550.jpg?1729557072" title=" <strong>Figure 1</strong><br/> <p>Two-step “hydantoinase process” for production of D-amino acids.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/40'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g002-550.jpg?1729557075" title=" <strong>Figure 2</strong><br/> <p>(<b>A</b>) SDS-PAGE of recombinant <span class="html-italic">E. coli</span> BL21 Star/pET24a(+)-<span class="html-italic">PsDcase</span> cells. Lines 1–3 represent proteins induced for 6, 8, and 20 h in crude extract; line 4—protein markers (ROTI<sup>®</sup>Mark TRICOLOR); lines 5–7—proteins of the supernatant fraction of the same samples; lines 8–10—proteins in the precipitated insoluble fraction of the same samples. (<b>B</b>) SDS-PAGE of D-carbamoylase purified with Ni-NTA protocol. Line 1—crude extract induced for 20 h; line 2—protein markers (Thermo Scientific<sup>TM</sup> PageRuler<sup>TM</sup>); line 3—partially purified protein; line 4—purified protein (93.6%).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/40'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g003-550.jpg?1729557078" title=" <strong>Figure 3</strong><br/> <p>Docking model of the interaction of <span class="html-italic">Pseudomonas</span> sp. KNK003A D-carbamoylase with N-carbamoyl-D-tryptophan in the substrate-binding pocket. (<b>A</b>) Two-dimensional image of molecular interactions in the enzyme. (<b>B</b>) Three-dimensional image of molecular interactions in the enzyme. Structure with green and purple dotted lines shows hydrogen bonds and hydrophobic interactions and their lengths in angstroms, respectively. The surface indicates that the binding pocket is mainly hydrophilic along with hydrophobic regions.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/40'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g004-550.jpg?1729557080" title=" <strong>Figure 4</strong><br/> <p>Monitoring RMSD (<b>A</b>) for D-carbamoylase alone (blue), for the D-carbamoylase/N-carbamoyl-D-tryptophan complex (green), and for N-carbamoyl-D-tryptophan (orange) and RMSF (<b>B</b>) for D-carbamoylase alone (blue) and for the complex (green), as well as RMSF (<b>C</b>) of catalytically important amino acids over a 100 ns simulation.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/40'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g005-550.jpg?1729557082" title=" <strong>Figure 5</strong><br/> <p>Monitoring of the radius of gyration (<b>A</b>) and monitoring of the SASA (<b>B</b>) in protein (blue) and protein–ligand complex (green) during 100 ns simulation.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/40'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00040/article_deploy/html/images/biotech-13-00040-g006-550.jpg?1729557084" title=" <strong>Figure 6</strong><br/> <p>Number of H bonds throughout the simulation time of 100 ns between protein and ligand (<b>A</b>), Arg176 and ligand (<b>B</b>), Asn173 and ligand (<b>C</b>). Blue bars indicate the number of bonds formed during each ns.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/40'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1491657" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 19 pages, 34131 KiB   </span> <a href="/2673-6284/13/4/39/pdf?version=1728264439" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Optimization of Sugar Extraction Process from Date Waste Using Full Factorial Design Toward Its Use for New Biotechnological Applications" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <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> <a class="title-link" href="/2673-6284/13/4/39">Optimization of Sugar Extraction Process from Date Waste Using Full Factorial Design Toward Its Use for New Biotechnological Applications</a> <div class="authors"> by <span class="inlineblock "><strong>Islam Sayah</strong>, </span><span class="inlineblock "><strong>Mondher Njehi</strong>, </span><span class="inlineblock "><strong>Nicola Cicero</strong>, </span><span class="inlineblock "><strong>Vincenzo Nava</strong>, </span><span class="inlineblock "><strong>Manel Ben M’hadheb</strong>, </span><span class="inlineblock "><strong>Hatem Majdoub</strong>, </span><span class="inlineblock "><strong>Sami Achour</strong> and </span><span class="inlineblock "><strong>Teresa Gervasi</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 39; <a href="https://doi.org/10.3390/biotech13040039">https://doi.org/10.3390/biotech13040039</a> - 3 Oct 2024 </div> Viewed by 1103 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> In Tunisia, the date industry generates a large quantity of waste, raising environmental concerns. However, dates are rich in sugars, which offer a renewable source of nutrients for various applications. In this study, sugar extraction from two low-grade pitted date fruits (Alig and <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/39/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> In Tunisia, the date industry generates a large quantity of waste, raising environmental concerns. However, dates are rich in sugars, which offer a renewable source of nutrients for various applications. In this study, sugar extraction from two low-grade pitted date fruits (Alig and Kentichi) under ultrasound, was optimized using full factorial design. At 40 °C, for20 min, and with a liquid-to-solid ratio of 10 mL/g, the optimum sugar contents were 60.87% and 50.79% for the varieties Alig and Kentichi, respectively. The date extracts were chemically analyzed, revealing low fat and protein contents, but significant polyphenol and mineral contents in both varieties. HPLC-IR analysis revealed more inverted sugars (glucose and fructose) in the Alig variety and more sucrose in the Kentichi variety. FTIR and SEM analysis showed the efficiency of the ultrasonic treatment of the biomass in terms of improving mass transfer diffusion through ultrasonic cavitation. Thus, ultrasound-assisted extraction constitutes an effective method for the recovery of sugar from date waste. <a href="/2673-6284/13/4/39">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Section <a href="/journal/biotech/sections/agricultural_food_biotechnology">Agricultural and Food Biotechnology</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/39/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1491657"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1491657"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1491657" data-cycle-prev="#prev1491657" data-cycle-progressive="#images1491657" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1491657-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g001-550.jpg?1728264613" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1491657" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1491657-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g002-550.jpg?1728264617'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1491657-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g003-550.jpg?1728264619'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1491657-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g004-550.jpg?1728264620'><p>Figure 4</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1491657-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g005-550.jpg?1728264624'><p>Figure 5</p></div> --- <div class='openpopupgallery' data-imgindex='5' data-target='article-1491657-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g006-550.jpg?1728264625'><p>Figure 6</p></div> --- <div class='openpopupgallery' data-imgindex='6' data-target='article-1491657-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g007-550.jpg?1728264627'><p>Figure 7</p></div> --- <div class='openpopupgallery' data-imgindex='7' data-target='article-1491657-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g008-550.jpg?1728264636'><p>Figure 8</p></div> --- <div class='openpopupgallery' data-imgindex='8' data-target='article-1491657-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g009-550.jpg?1728264643'><p>Figure 9</p></div></script></div></div><div id="article-1491657-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g001-550.jpg?1728264613" title=" <strong>Figure 1</strong><br/> <p>Pareto chart of the standardized effects (<b>a</b>); main effects plot for the variety Alig (<b>b</b>).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/39'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g002-550.jpg?1728264617" title=" <strong>Figure 2</strong><br/> <p>Three-dimensional surface plots and two-dimensional contour plots for the variety Alig. ((<b>a</b>,<b>b</b>) extraction temperature and time; (<b>c</b>,<b>d</b>) extraction temperature and liquid-to-solid ratio, and (<b>e</b>,<b>f</b>) extraction time and liquid-to-solid ratio).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/39'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g003-550.jpg?1728264619" title=" <strong>Figure 3</strong><br/> <p>Optimal extraction conditions for Alig variety.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/39'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g004-550.jpg?1728264620" title=" <strong>Figure 4</strong><br/> <p>Pareto chart of the standardized effects (<b>a</b>); main effects plot for the variety Kentichi (<b>b</b>).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/39'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g005-550.jpg?1728264624" title=" <strong>Figure 5</strong><br/> <p>Three-dimensional surface plots and two-dimensional contour plots for the variety Kentichi ((<b>a</b>,<b>b</b>) temperature versus time; (<b>c</b>,<b>d</b>) temperature versus liquid-to-solid ratio; and (<b>e</b>,<b>f</b>) time versus liquid-to-solid ratio).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/39'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g006-550.jpg?1728264625" title=" <strong>Figure 6</strong><br/> <p>Optimal extraction conditions for Kentichi variety.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/39'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g007-550.jpg?1728264627" title=" <strong>Figure 7</strong><br/> <p>FT-IR spectra of Alig fruit (AF), Alig residue (AR), and Alig liquid extract (AL) (<b>a</b>,<b>b</b>), as well as Kentichi fruit (KF), Kentichi residue (KR), and Kentichi liquid extract (KL) (<b>c</b>,<b>d</b>).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/39'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g008-550.jpg?1728264636" title=" <strong>Figure 8</strong><br/> <p>SEM images with different magnifications of Alig fruit (AF) (<b>a</b>,<b>c</b>,<b>e</b>), Alig residue (AR) (<b>b</b>,<b>d</b>,<b>f</b>).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/39'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00039/article_deploy/html/images/biotech-13-00039-g009-550.jpg?1728264643" title=" <strong>Figure 9</strong><br/> <p>SEM images with different magnifications of Kentichi fruit (KF) (<b>a</b>,<b>c</b>,<b>e</b>), and Kentichi residue (KR) (<b>b</b>,<b>d</b>,<b>f</b>).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/39'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1485704" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 18 pages, 1912 KiB   </span> <a href="/2673-6284/13/4/38/pdf?version=1727411972" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Chemical, Textural and Antioxidant Properties of Oat-Fermented Beverages with Different Starter Lactic Acid Bacteria and Pectin" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <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> <a class="title-link" href="/2673-6284/13/4/38">Chemical, Textural and Antioxidant Properties of Oat-Fermented Beverages with Different Starter Lactic Acid Bacteria and Pectin</a> <div class="authors"> by <span class="inlineblock "><strong>Dmitrii V. Khrundin</strong> and </span><span class="inlineblock "><strong>Elena V. Nikitina</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(4), 38; <a href="https://doi.org/10.3390/biotech13040038">https://doi.org/10.3390/biotech13040038</a> - 25 Sep 2024 </div> Viewed by 649 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Currently, starter cultures for fermenting plant-based beverages are not widely available commercially, but producers can use starter cultures for dairy products. Therefore, the aim of this study was to determine the physicochemical, rheological, antioxidant and sensory properties of oat beverages with/without pectin fermented <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/38/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Currently, starter cultures for fermenting plant-based beverages are not widely available commercially, but producers can use starter cultures for dairy products. Therefore, the aim of this study was to determine the physicochemical, rheological, antioxidant and sensory properties of oat beverages with/without pectin fermented by four different dairy starter cultures. The use of a mono-starter with <i>Lactobacillus bulgaricus</i> or <i>Sreptococcus thermophilus</i> allows for the efficient use of glucose, and more lactic acid is accumulated. The beverage with <i>L. bulgaricus</i> is characterised by high adhesion, syneresis and low cohesiveness, and it has high antioxidant activity and a low sensory profile. Using starter with <i>L. bulgaricus</i>, <i>S. thermophilus</i> and some <i>Lactococcus</i> for fermentation yields a product with high sensory capacity, forming a high-viscosity beverage matrix with low syneresis, high water retention, chewy texture and stickiness. It has been observed that the absence of lactococci and the presence of <i>Lactobacillus casei</i>, <i>L. Rhamnosus</i> and <i>L. paracasei</i> in the starter yields a product with high antioxidant activity, especially in the presence of pectin. The use of pectin significantly improves the viscosity and textural properties of oat yoghurt, enhancing the drink’s flavour and giving it body. For many reasons, the use of different commercial starters in the dairy industry results in different viscosities of oat fermented beverages, forming a matrix with different textural, sensory and antioxidant properties. <a href="/2673-6284/13/4/38">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Section <a href="/journal/biotech/sections/agricultural_food_biotechnology">Agricultural and Food Biotechnology</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/4/38/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1485704"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1485704"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1485704" data-cycle-prev="#prev1485704" data-cycle-progressive="#images1485704" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1485704-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g001-550.jpg?1727412081" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1485704" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1485704-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g002-550.jpg?1727412083'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1485704-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g003-550.jpg?1727412085'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1485704-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g004-550.jpg?1727412085'><p>Figure 4</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1485704-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g005-550.jpg?1727412088'><p>Figure 5</p></div> --- <div class='openpopupgallery' data-imgindex='5' data-target='article-1485704-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g006-550.jpg?1727412089'><p>Figure 6</p></div> --- <div class='openpopupgallery' data-imgindex='6' data-target='article-1485704-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g007-550.jpg?1727412090'><p>Figure 7</p></div></script></div></div><div id="article-1485704-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g001-550.jpg?1727412081" title=" <strong>Figure 1</strong><br/> <p>The production steps of the fermented oat base.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/38'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g002-550.jpg?1727412083" title=" <strong>Figure 2</strong><br/> <p>Effect of starters and pectin on pH (<b>A</b>) and titratable acidity (<b>B</b>), absolute glucose concentration (<b>C</b>) and percentage of residual glucose (<b>D</b>). Blue bar—unfermented oat base without pectin, red bar—unfermented oat base with pectin, grey bar—oat-based fermented beverage without pectin, black bar—oat-based fermented beverage with pectin. The blue line shows the level of the index for the oat base without pectin and the red line shows the level of the index for the oat base with pectin. Asterisks indicate statistically significant differences between variants without pectin (control) and with pectin according to non-parametric one-way analysis of variance (Kruskal–Wallis) test, <span class="html-italic">p</span> &lt; 0.05.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/38'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g003-550.jpg?1727412085" title=" <strong>Figure 3</strong><br/> <p>Effect of starter and the pectin to concentration on total phenol-containing compounds (TPCs) ((<b>A</b>), product; (<b>B</b>), protein-free extract). Blue bar—unfermented oat base without pectin; red bar—unfermented oat base with pectin; grey bar—oat-based fermented beverage without pectin; black bar—oat-based fermented beverage with pectin. The blue line shows the level of the index for the oat base without pectin and the red line shows the level of the index for the oat base with pectin. Asterisks indicate statistically significant differences between variants without pectin (control) and with pectin according to non-parametric one-way analysis of variance (Kruskal–Wallis) test, <span class="html-italic">p</span> &lt; 0.05.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/38'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g004-550.jpg?1727412085" title=" <strong>Figure 4</strong><br/> <p>Total amount of extractable polysaccharides (EPSs). Blue bar—unfermented oat base without pectin; red bar—unfermented oat base with pectin; grey bar—oat-based fermented beverage without pectin; black bar—oat-based fermented beverage with pectin. The blue line shows the level of the index for the oat base without pectin and the red line shows the level of the index for the oat base with pectin. Asterisks indicate statistically significant differences between variants without pectin (control) and with pectin according to non-parametric one-way analysis of variance (Kruskal–Wallis) test, <span class="html-italic">p</span> &lt; 0.05.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/38'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g005-550.jpg?1727412088" title=" <strong>Figure 5</strong><br/> <p>Sensory profile of oat-based fermented beverage. Controls without pectin (LB, ST, ProBio, Symb and oat base) are indicated by the thick solid lines, samples with pectin (LB + pectin, ST + pectin, ProBio + pectin, Symb + pectin and oat base + pectin) are indicated by the dashed lines.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/38'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g006-550.jpg?1727412089" title=" <strong>Figure 6</strong><br/> <p>Antioxidant activity of fermented oat base: Ferric-reducing antioxidant power (<b>A</b>); Radical-scavenging ability (<b>B</b>); OH free-radical-scavenging ability (<b>C</b>). Blue bar—unfermented oat base without pectin, red bar—unfermented oat base with pectin, grey bar—oat-based fermented beverage without pectin, black bar—oat-based fermented beverage with pectin. The blue line shows the level of the index for the oat base without pectin and the red line shows the level of the index for the oat base with pectin. Asterisks indicate statistically significant differences between variants without pectin (control) and with pectin according to non-parametric one-way analysis of variance (Kruskal–Wallis) test, <span class="html-italic">p</span> &lt; 0.05.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/38'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00038/article_deploy/html/images/biotech-13-00038-g007-550.jpg?1727412090" title=" <strong>Figure 7</strong><br/> <p>Principal component analysis (<b>A</b>) of oat base and oat-based fermented beverages with/without pectin, dendrogram of oat-based fermented beverages with/without pectin (<b>B</b>) and heat-map correlation (<b>C</b>) of antioxidant activity and chemical composition.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/4/38'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1483590" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 18 pages, 3899 KiB   </span> <a href="/2673-6284/13/3/37/pdf?version=1727085746" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="The Elimination of Viroids through In Vitro Thermotherapy and a Meristem Tip Culture from a New Limonime Hybrid (Citrus x limon var. limon (L.) Burm. f. x Citrus latifolia var. latifolia)" data-journal="biotech"> <i class="material-icons custom-download"></i> </a> </div> <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> <a class="title-link" href="/2673-6284/13/3/37">The Elimination of Viroids through In Vitro Thermotherapy and a Meristem Tip Culture from a New Limonime Hybrid (<i>Citrus</i> x <i>limon</i> var. <i>limon</i> (L.) Burm. f. x <i>Citrus latifolia</i> var. <i>latifolia</i>)</a> <div class="authors"> by <span class="inlineblock "><strong>Virginia Sarropoulou</strong>, </span><span class="inlineblock "><strong>Katerina Grigoriadou</strong>, </span><span class="inlineblock "><strong>Varvara I. Maliogka</strong>, </span><span class="inlineblock "><strong>Chrysoula-Lito Sassalou</strong> and </span><span class="inlineblock "><strong>Vasileios Ziogas</strong></span> </div> <div class="color-grey-dark"> <em>BioTech</em> <b>2024</b>, <em>13</em>(3), 37; <a href="https://doi.org/10.3390/biotech13030037">https://doi.org/10.3390/biotech13030037</a> - 23 Sep 2024 </div> Viewed by 787 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Viruses and viroids pose a significant challenge in citriculture, and their control is crucial for plant health. This study evaluated the effectiveness of in vitro thermotherapy combined with a meristem tip culture for eliminating citrus exocortis viroid (CEVd) and hop stunt viroid (HSVd) <a href="#" data-counterslink = "https://www.mdpi.com/2673-6284/13/3/37/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Viruses and viroids pose a significant challenge in citriculture, and their control is crucial for plant health. This study evaluated the effectiveness of in vitro thermotherapy combined with a meristem tip culture for eliminating citrus exocortis viroid (CEVd) and hop stunt viroid (HSVd) from a new limonime hybrid (<i>Citrus</i> x <i>limon</i> var. <i>limon</i> x <i>Citrus latifolia</i> var. <i>latifolia</i>). The elimination success was confirmed by RT-PCR assays. The in vitro elimination rate for CEVd during the shoot proliferation stage (43%) was higher than for HSVd (21%). Accordingly, in the subsequent rooting stage, the in vitro elimination rate for CEVd (50%) was higher than for HSVd (33%). Successful CEVd and HSVd eradication at a 100% rate was confirmed in the ex vitro acclimatized plants in the greenhouse. The study also established an efficient micropropagation protocol. The optimal treatment for in vitro shoot induction was 0.5–2 mg L<sup>−1</sup> benzyladenine (BA) + 0.5 mg L<sup>−1</sup> gibberellic acid (GA<sub>3</sub>) + 0.25 mg L<sup>−1</sup> naphthalene acetic acid (NAA), while for shoot elongation, it was 0.5 mg L<sup>−1</sup> BA + 0.5 mg L<sup>−1</sup> kinetin (KIN) + 0.5 mg L<sup>−1</sup> GA<sub>3</sub> + 0.25 mg L<sup>−1</sup> NAA. Rooting was best promoted by 1 mg L<sup>−1</sup> NAA. This study provides valuable insights for the mass production of viroid-free propagation material in this new lemon x lime hybrid, contributing to the conservation of genetic resources in citrus breeding programs through the combined application of in vitro thermotherapy and an in vitro meristem tip culture, a novel and highlighted achievement reported for the first time in this study. <a href="/2673-6284/13/3/37">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Section <a href="/journal/biotech/sections/agricultural_food_biotechnology">Agricultural and Food Biotechnology</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2673-6284/13/3/37/show" ><span >►</span><span style=" display: none;">▼</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1483590"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1483590"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1483590" data-cycle-prev="#prev1483590" data-cycle-progressive="#images1483590" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1483590-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g001-550.jpg?1727085868" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1483590" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1483590-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g002-550.jpg?1727085870'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1483590-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g003-550.jpg?1727085872'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1483590-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g004-550.jpg?1727085875'><p>Figure 4</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1483590-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g005-550.jpg?1727085877'><p>Figure 5</p></div> --- <div class='openpopupgallery' data-imgindex='5' data-target='article-1483590-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g006-550.jpg?1727085878'><p>Figure 6</p></div> --- <div class='openpopupgallery' data-imgindex='6' data-target='article-1483590-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g007-550.jpg?1727085883'><p>Figure 7</p></div></script></div></div><div id="article-1483590-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g001-550.jpg?1727085868" title=" <strong>Figure 1</strong><br/> <p>In vitro shoot proliferation and/or rooting of <span class="html-italic">Citrus</span> x <span class="html-italic">limon</span> var. <span class="html-italic">limon</span> (L.) Burm. f. x <span class="html-italic">Citrus latifolia</span> var. <span class="html-italic">latifolia</span>) after 30 days of culture in modified MS (x 2FeEDTA) medium enriched with 30 g L<sup>−1</sup> sucrose and 6 g L<sup>−1</sup> Plant Agar under different types (ΒA, ΚΙΝ) and concentrations (0, 0.5, 1, 2 mg L<sup>−1</sup>) of cytokinins applied individually and combined with 0.5 mg L<sup>−1</sup> GA<sub>3</sub> + 0.25 mg L<sup>−1</sup> NAA: (<b>a</b>) inside culture vessels; (<b>b</b>) outside culture vessels.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/3/37'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g002-550.jpg?1727085870" title=" <strong>Figure 2</strong><br/> <p>The in vitro rooting and vegetative growth of <span class="html-italic">Citrus</span> x <span class="html-italic">limon</span> var. <span class="html-italic">limon</span> (L.) Burm. f. x <span class="html-italic">Citrus latifolia</span> var. <span class="html-italic">latifolia</span> after 30 days of culture in a modified MS (x2FeEDTA) nutrient medium enriched with 30 g L<sup>−1</sup> sucrose and 6 g L<sup>−1</sup> Plant Agar under the effect of different types (IBA, NAA, IAA) and concentrations (0, 0.5, 1, 2 mg L<sup>−1</sup>) of auxins: (<b>a</b>) inside culture vessels; (<b>b</b>) outside culture vessels.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/3/37'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g003-550.jpg?1727085872" title=" <strong>Figure 3</strong><br/> <p>The ex vitro acclimatization of in vitro rooted plants to the greenhouse and further vegetative growth of plants in a 6-month period from late spring to mid-winter (consecutive transplants into larger pots, substrate mixture growth-dependent) in <span class="html-italic">C.</span> x <span class="html-italic">limon</span> var. <span class="html-italic">limon</span> (L.) Burm. f. x <span class="html-italic">C. latifolia</span> var. <span class="html-italic">latifolia</span>: (<b>a</b>) the in vitro rooted plant; (<b>b</b>) the ex vitro acclimatized plant in a 0.33 L pot; (<b>c</b>) the growth of the acclimatized plant in a 2.5 L pot.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/3/37'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g004-550.jpg?1727085875" title=" <strong>Figure 4</strong><br/> <p>Vegetative growth and development and appearance of stress symptoms in shoot-tip explants during in vitro thermotherapy at successively increasing temperature regimes in <span class="html-italic">Citrus</span> x <span class="html-italic">limon</span> var. <span class="html-italic">limon</span> (L.) Burm. f. x <span class="html-italic">Citrus latifolia</span> var. <span class="html-italic">latifolia</span>.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/3/37'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g005-550.jpg?1727085877" title=" <strong>Figure 5</strong><br/> <p>In vitro differentiation and development of apical meristems (0.5–1 mm) into new complete shoot tips in modified MS (x 2Fe) medium enriched with 30 g L<sup>−1</sup> sucrose, 0.5 mg L<sup>−1</sup> BA, 0.5 mg L<sup>−1</sup> KIN, 0.5 mg L<sup>−1</sup> GA<sub>3</sub>, 0.25 mg L<sup>−1</sup> NAA (pH 5.8), and 6 g L<sup>−1</sup> Plant Agar in <span class="html-italic">Citrus</span> x <span class="html-italic">limon</span> var. <span class="html-italic">limon</span> (L.) Burm. f. x <span class="html-italic">Citrus latifolia</span> var. <span class="html-italic">latifolia</span>: (<b>a</b>) growth of apical meristem after one week of culture; (<b>b</b>) after 30 days; (<b>c</b>) after 60 days.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/3/37'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g006-550.jpg?1727085878" title=" <strong>Figure 6</strong><br/> <p>Plant material samples tested by RT-PCR for CEVd and HSVd in <span class="html-italic">Citrus</span> x <span class="html-italic">limon</span> var. <span class="html-italic">limon</span> (L.) Burm. f. x <span class="html-italic">Citrus latifolia</span> var. <span class="html-italic">latifolia</span>: (<b>a</b>) 14 in vitro shoot tips cultured in MS (x 2Fe) + 30 g L<sup>−1</sup> sucrose + 0.5 mg L<sup>−1</sup> BA + 0.5 mg L<sup>−1</sup> KIN + 0.5 mg L<sup>−1</sup> GA<sub>3</sub> + 0.25 mg L<sup>−1</sup> NAA (pH 5.8) + 6 g L<sup>−1</sup> Plant Agar (1st round RT-PCR); (<b>b</b>) 6 in vitro rooted shoot tips cultured in MS (x 2Fe) + 30 g L<sup>−1</sup> sucrose + 1 mg L<sup>−1</sup> NAA (pH 5.8) + 6 g L<sup>−1</sup> Plant Agar (2nd round RT-PCR); (<b>c</b>) 6-month ex vitro acclimatized greenhouse plant sample (code: Lime 16.3) (3rd round RT-PCR); (<b>d</b>) subsequent vegetative growth of acclimatized viroid-free plant under greenhouse conditions during late spring–mid-winter.</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/3/37'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/biotech/biotech-13-00037/article_deploy/html/images/biotech-13-00037-g007-550.jpg?1727085883" title=" <strong>Figure 7</strong><br/> <p>The agarose gel electrophoretic analysis of RT-PCR products obtained using (<b>A</b>) the primers (22) targeting the ubiquitin gene (internal control) [<a href="#B33-biotech-13-00037" class="html-bibr">33</a>]; (<b>B</b>) the primers targeting the HSVd [<a href="#B32-biotech-13-00037" class="html-bibr">32</a>]; (<b>C</b>) The primers targeting CEVd [<a href="#B34-biotech-13-00037" class="html-bibr">34</a>]; (<b>D</b>) the agarose gel electrophoretic analysis of the 3rd round of RT-PCR in the ex vitro acclimatized greenhouse sample plant (code: Lime 16.3) after thermotherapy and tissue culture rescue for HSVd (1) and CEVd (2).</p> <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2673-6284/13/3/37'>Full article</a></strong> "></a></div> </div> </div> <span class="more" 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