Anh T Tran, Junhao Wen, Gaby Abou Karam, Dorin Zeevi, Adnan I Qureshi, Ajay Malhotra, Shahram Majidi, Niloufar Valizadeh, Santosh B Murthy, Mert R Sabuncu, David Roh, Guido J Falcone, Kevin N Sheth, Seyedmehdi Payabvash
Handcrafted radiomics use predefined formulas to extract quantitative features from medical images, whereas deep neural networks learn de novo features through iterative training. We compared these approaches for predicting 3-month outcomes and hematoma expansion from admission non-contrast head CT in acute intracerebral hemorrhage (ICH). Training and cross-validation were performed using a multicenter trial cohort (n = 866), with external validation on a single-center dataset (n = 645). We trained multiscale U-shaped segmentation models for hematoma segmentation and extracted (i) radiomics from the segmented lesions and (ii) two latent deep feature sets-from the segmentation encoder and a generative autoencoder trained on dilated lesion patches. Features were reduced with unsupervised Non-Negative Matrix Factorization (NMF) to 128 per set and used-alone or in combination-for six machine-learning classifiers to predict 3-month clinical outcomes and (>3, >6, >9 mL) hematoma expansion thresholds. The addition of latent deep features to radiomics numerically increased model prediction performance for 3-month outcomes and hematoma expansion using Random Forest, XGBoost, Extra Trees, or Elastic Net classifiers; however, the improved accuracy only reached statistical significance in predicting >3 mL hematoma expansion. Clinically, these consistent but modest increases in prediction performance may improve risk stratification at the individual level. Nevertheless, the latent deep features show potential for extracting additional clinically relevant information from admission head CT for prognostication in hemorrhagic stroke.
{"title":"Comparing Handcrafted Radiomics Versus Latent Deep Learning Features of Admission Head CT for Hemorrhagic Stroke Outcome Prediction.","authors":"Anh T Tran, Junhao Wen, Gaby Abou Karam, Dorin Zeevi, Adnan I Qureshi, Ajay Malhotra, Shahram Majidi, Niloufar Valizadeh, Santosh B Murthy, Mert R Sabuncu, David Roh, Guido J Falcone, Kevin N Sheth, Seyedmehdi Payabvash","doi":"10.3390/biotech14040087","DOIUrl":"10.3390/biotech14040087","url":null,"abstract":"<p><p>Handcrafted radiomics use predefined formulas to extract quantitative features from medical images, whereas deep neural networks learn de novo features through iterative training. We compared these approaches for predicting 3-month outcomes and hematoma expansion from admission non-contrast head CT in acute intracerebral hemorrhage (ICH). Training and cross-validation were performed using a multicenter trial cohort (n = 866), with external validation on a single-center dataset (n = 645). We trained multiscale U-shaped segmentation models for hematoma segmentation and extracted (i) radiomics from the segmented lesions and (ii) two latent deep feature sets-from the segmentation encoder and a generative autoencoder trained on dilated lesion patches. Features were reduced with unsupervised Non-Negative Matrix Factorization (NMF) to 128 per set and used-alone or in combination-for six machine-learning classifiers to predict 3-month clinical outcomes and (>3, >6, >9 mL) hematoma expansion thresholds. The addition of latent deep features to radiomics numerically increased model prediction performance for 3-month outcomes and hematoma expansion using Random Forest, XGBoost, Extra Trees, or Elastic Net classifiers; however, the improved accuracy only reached statistical significance in predicting >3 mL hematoma expansion. Clinically, these consistent but modest increases in prediction performance may improve risk stratification at the individual level. Nevertheless, the latent deep features show potential for extracting additional clinically relevant information from admission head CT for prognostication in hemorrhagic stroke.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 4","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12641684/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145589196","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Laura Martínez-Alarcón, Sergio Liarte, Juana M Abellaneda, Juan J Quereda, Livia Mendonça, Antonio Muñoz, Pablo Ramírez, Guillermo Ramis
Cell transplantation is often performed with ultrasonographic guidance for accurate delivery through injection. In such procedures, using ultrasonographic contrast greatly improves target delivery. However, accumulating evidence suggests that exposure to such contrast agents may have negative effects on transplanted cells. No study so far has researched this issue. Stabilized sulfur hexafluoride (SF6) microbubbles are a widely used sonographic contrast agent. Skin hCD55 porcine transgenic fibroblasts and mesenchymal stem cells from human bone marrow (hMSCs) were exposed in vitro to SF6 in concentrations ranging from 1.54 µM to 308 µM. The effects on viability and cell growth were registered using an impedance-based label-free Real-Time Cell Analyzer (RTCA). Data was recorded every 15 min for 50 h of total study time. Both cell lines behave distinctly when exposed to SF6. Porcine fibroblast growth showed relevant alterations only when exposed to higher concentrations. In contrast, hMSCs showed progressive growth decrease in relation to SF6 concentration. Taken together, while SF6-based contrast agents pose no threat to patient safety, our results indicate that exposure of suspended stem cells to the contrast agent could affect the effective dose administered in cell therapy procedures. This prompts specific cell lineage testing, adjusting methods and properly compensating for cell loss, with a potential impact on procedural cost and success rates.
{"title":"Exposure to Sulfur Hexafluoride Influences Viability in Cell Transplant Suspensions.","authors":"Laura Martínez-Alarcón, Sergio Liarte, Juana M Abellaneda, Juan J Quereda, Livia Mendonça, Antonio Muñoz, Pablo Ramírez, Guillermo Ramis","doi":"10.3390/biotech14040086","DOIUrl":"10.3390/biotech14040086","url":null,"abstract":"<p><p>Cell transplantation is often performed with ultrasonographic guidance for accurate delivery through injection. In such procedures, using ultrasonographic contrast greatly improves target delivery. However, accumulating evidence suggests that exposure to such contrast agents may have negative effects on transplanted cells. No study so far has researched this issue. Stabilized sulfur hexafluoride (SF6) microbubbles are a widely used sonographic contrast agent. Skin hCD55 porcine transgenic fibroblasts and mesenchymal stem cells from human bone marrow (hMSCs) were exposed in vitro to SF6 in concentrations ranging from 1.54 µM to 308 µM. The effects on viability and cell growth were registered using an impedance-based label-free Real-Time Cell Analyzer (RTCA). Data was recorded every 15 min for 50 h of total study time. Both cell lines behave distinctly when exposed to SF6. Porcine fibroblast growth showed relevant alterations only when exposed to higher concentrations. In contrast, hMSCs showed progressive growth decrease in relation to SF6 concentration. Taken together, while SF6-based contrast agents pose no threat to patient safety, our results indicate that exposure of suspended stem cells to the contrast agent could affect the effective dose administered in cell therapy procedures. This prompts specific cell lineage testing, adjusting methods and properly compensating for cell loss, with a potential impact on procedural cost and success rates.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 4","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12641703/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145589181","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xylooligosaccharides (XOS) are functional oligosaccharides with recognized prebiotic properties and growing industrial relevance, typically obtained through enzymatic depolymerization of xylan-rich lignocellulosic substrates. In this study, a recombinant endo-β-1,4-xylanase (XynA) from Clostridium acetobutylicum was employed for XOS production. The xynA gene was cloned into the expression vector pET-21a(+) and heterologously expressed in Escherichia coli BL21(DE3) under induction with isopropyl β-D-1-thiogalactopyranoside (IPTG). The recombinant protein, with an estimated molecular mass of 37.5 kDa, was verified by SDS-PAGE and Western blot analysis. Functional characterization via thin-layer chromatography revealed that XynA efficiently hydrolyzed beechwood xylan and rye arabinoxylan, predominantly yielding xylobiose. Additionally, the enzyme catalyzed the conversion of xylotriose into xylobiose and trace amounts of xylose. Notably, XynA demonstrated hydrolytic activity against autohydrolysed and alkali-pretreated coconut husk biomass, facilitating the release of XOS. These results underscore the potential of C. acetobutylicum XynA as a biocatalyst for the valorization of lignocellulosic residues into high-value oligosaccharides.
低聚木糖(XOS)是一种功能性低聚糖,具有公认的益生元特性和日益增长的工业相关性,通常通过酶解聚合获得富含木聚糖的木质纤维素底物。本研究利用乙酰丁酸梭菌(Clostridium acetobutylicum)的重组内切-β-1,4-木聚糖酶(XynA)生产XOS。将xynA基因克隆到表达载体pET-21a(+)中,在异丙基β- d -1-硫代半乳糖苷(IPTG)诱导下,在大肠杆菌BL21(DE3)中异源表达。重组蛋白经SDS-PAGE和Western blot验证,分子量约为37.5 kDa。薄层色谱功能表征表明,XynA能有效水解山毛榉木聚糖和黑麦阿拉伯木聚糖,主要产木糖糖。此外,该酶还催化木糖三糖转化为木糖二糖和微量木糖。值得注意的是,XynA对自水解和碱预处理的椰子壳生物质具有水解活性,促进了XOS的释放。这些结果强调了C. acetobutylicum XynA作为木质纤维素残基转化为高价值低聚糖的生物催化剂的潜力。
{"title":"Recombinant <i>Clostridium acetobutylicum</i> Endoxylanase for Xylooligosaccharide Production from Pretreated Lignocellulosic Biomass.","authors":"Afifa Husna, Agustin Krisna Wardani, Chun-Yi Hu, Yo-Chia Chen","doi":"10.3390/biotech14040085","DOIUrl":"10.3390/biotech14040085","url":null,"abstract":"<p><p>Xylooligosaccharides (XOS) are functional oligosaccharides with recognized prebiotic properties and growing industrial relevance, typically obtained through enzymatic depolymerization of xylan-rich lignocellulosic substrates. In this study, a recombinant endo-β-1,4-xylanase (XynA) from <i>Clostridium acetobutylicum</i> was employed for XOS production. The xynA gene was cloned into the expression vector pET-21a(+) and heterologously expressed in <i>Escherichia coli</i> BL21(DE3) under induction with isopropyl β-D-1-thiogalactopyranoside (IPTG). The recombinant protein, with an estimated molecular mass of 37.5 kDa, was verified by SDS-PAGE and Western blot analysis. Functional characterization via thin-layer chromatography revealed that XynA efficiently hydrolyzed beechwood xylan and rye arabinoxylan, predominantly yielding xylobiose. Additionally, the enzyme catalyzed the conversion of xylotriose into xylobiose and trace amounts of xylose. Notably, XynA demonstrated hydrolytic activity against autohydrolysed and alkali-pretreated coconut husk biomass, facilitating the release of XOS. These results underscore the potential of <i>C. acetobutylicum</i> XynA as a biocatalyst for the valorization of lignocellulosic residues into high-value oligosaccharides.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 4","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12641949/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145588216","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Julian Tix, Fernando Pedraza, Roland Ulber, Nils Tippkötter
Carbon capture and power-to-X are becoming increasingly relevant in the context of decarbonization and supply security. Actinobacillus succinogenes is capable of transforming CO2 into succinate, whereby product formation is significantly limited by the availability of NADH. The aim of this work was to further develop a bioelectrochemical system (BES) in order to provide additional reduction equivalents and thus increase yield and titer. To this end, a new BES configuration was established. A conventional stirred tank reactor (STR) is coupled via a bypass to an H-cell, in which the redox mediator neutral red (NR) is electrochemically reduced and then returned back to the bioreactor. The indirect electron transfer decouples the electrochemical reduction from the biology and results in increased intracellular availability of NADH. The present approach resulted in an increase in yield from 0.64 g·g-1 to 0.76 g·g-1, corresponding to an increase of 18%. At the same time, a titer of 16.48 ± 0.19 g·L-1 was achieved in the BES, compared to 12.05 ± 0.18 g·L-1 in the control. The results show that the mediator-assisted, partially decoupled BES architecture significantly improves CO2-based succinate production and opens up a scalable path to the use of renewable electricity as a reduction source in power-to-X processes.
{"title":"Enhanced Succinate Production in <i>Actinobacillus succinogenes</i> via Neutral Red Bypass Reduction in a Novel Bioelectrochemical System.","authors":"Julian Tix, Fernando Pedraza, Roland Ulber, Nils Tippkötter","doi":"10.3390/biotech14040084","DOIUrl":"10.3390/biotech14040084","url":null,"abstract":"<p><p>Carbon capture and power-to-X are becoming increasingly relevant in the context of decarbonization and supply security. <i>Actinobacillus succinogenes</i> is capable of transforming CO<sub>2</sub> into succinate, whereby product formation is significantly limited by the availability of NADH. The aim of this work was to further develop a bioelectrochemical system (BES) in order to provide additional reduction equivalents and thus increase yield and titer. To this end, a new BES configuration was established. A conventional stirred tank reactor (STR) is coupled via a bypass to an H-cell, in which the redox mediator neutral red (NR) is electrochemically reduced and then returned back to the bioreactor. The indirect electron transfer decouples the electrochemical reduction from the biology and results in increased intracellular availability of NADH. The present approach resulted in an increase in yield from 0.64 g·g<sup>-1</sup> to 0.76 g·g<sup>-1</sup>, corresponding to an increase of 18%. At the same time, a titer of 16.48 ± 0.19 g·L<sup>-1</sup> was achieved in the BES, compared to 12.05 ± 0.18 g·L<sup>-1</sup> in the control. The results show that the mediator-assisted, partially decoupled BES architecture significantly improves CO<sub>2</sub>-based succinate production and opens up a scalable path to the use of renewable electricity as a reduction source in power-to-X processes.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 4","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12641854/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145589225","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Suaeda salsa, an annual herb belonging to the genus Suaeda within the Chenopodiaceae family, is highly salt-tolerant and can thrive in large quantities on saline and alkaline soils. This study presents a novel fermentation technique to produce Suaeda tea, utilizing a synergistic blend of microbial agents: Kluyveromyces marxianus, Komagataeibacter europaeus, and Acetobacter schutzenbachii. The resulting tea demonstrates a potent antioxidant capacity, with a hydroxyl radical scavenging rate of 64.2% and an exceptional 1,1-diphenyl-2-picrylhydrazyl radical scavenging capacity of 83.3%, along with increased ferric ion reduction/antioxidant power (FRAP) reducing power (1.82), indicating its superior antioxidant profile. Through the comparison of different microbial strain combinations under varying process parameters such as fermentation temperature and duration, the experiment revealed that fermentation at 37 °C for 24 h results in the highest concentrations of tea polyphenols (TPs) (≥10.87 mg/mL) and free amino acids (26.32 mg/100 mL). The quality of the fermented Suaeda tea meets the stringent GB/T 21733-2008 standards for tea beverages, exhibiting excellent physicochemical indices and sensory attributes. The antioxidant efficacy of the fermented Suaeda tea persists significantly throughout a 180-day duration. The optimization of the fermentation process for Suaeda tea not only provides a theoretical framework for large-scale production but also establishes a foundation for Suaeda salsa in the tea beverage sector. This innovation enriches the market with a diverse range of health-promoting teas, catering to the growing consumer demand for nutritious and beneficial beverages.
{"title":"Preparation of Suaeda Tea Through Semi-Solid Fermentation Utilizing <i>Kluyveromyces marxianus</i>, <i>Komagataeibacter europaeus</i>, and <i>Acetobacter schutzenbachii</i>: Physicochemical Characteristics, Process Optimization, and Antioxidant Activity.","authors":"Aoqi Dong, Xiaoying Dong, Xinying Dai, Yanru Gao, Yuewen Ning, Xiya Fan, Haiyan Liu","doi":"10.3390/biotech14040083","DOIUrl":"10.3390/biotech14040083","url":null,"abstract":"<p><p><i>Suaeda salsa</i>, an annual herb belonging to the genus <i>Suaeda</i> within the <i>Chenopodiaceae</i> family, is highly salt-tolerant and can thrive in large quantities on saline and alkaline soils. This study presents a novel fermentation technique to produce Suaeda tea, utilizing a synergistic blend of microbial agents: <i>Kluyveromyces marxianus</i>, <i>Komagataeibacter europaeus</i>, and <i>Acetobacter schutzenbachii</i>. The resulting tea demonstrates a potent antioxidant capacity, with a hydroxyl radical scavenging rate of 64.2% and an exceptional 1,1-diphenyl-2-picrylhydrazyl radical scavenging capacity of 83.3%, along with increased ferric ion reduction/antioxidant power (FRAP) reducing power (1.82), indicating its superior antioxidant profile. Through the comparison of different microbial strain combinations under varying process parameters such as fermentation temperature and duration, the experiment revealed that fermentation at 37 °C for 24 h results in the highest concentrations of tea polyphenols (TPs) (≥10.87 mg/mL) and free amino acids (26.32 mg/100 mL). The quality of the fermented Suaeda tea meets the stringent GB/T 21733-2008 standards for tea beverages, exhibiting excellent physicochemical indices and sensory attributes. The antioxidant efficacy of the fermented Suaeda tea persists significantly throughout a 180-day duration. The optimization of the fermentation process for Suaeda tea not only provides a theoretical framework for large-scale production but also establishes a foundation for <i>Suaeda salsa</i> in the tea beverage sector. This innovation enriches the market with a diverse range of health-promoting teas, catering to the growing consumer demand for nutritious and beneficial beverages.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 4","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12641745/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145589241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Iridiam Hernández-Soto, Antonio Juárez-Maldonado, Alfredo Madariaga-Navarrete, Ashutosh Sharma, Antonio de Jesus Cenobio-Galindo, Jose Manuel Pinedo-Espinoza, Aracely Hernández-Pérez, Alma Delia Hernández-Fuentes
Argemone mexicana L. is considered a weed; however, it contains secondary metabolites that can control phytopathogenic fungi in vitro, with the potential to adapt its effectiveness in the field. In the present study, leaf extracts of A. mexicana (hexane and methanol) were prepared, and their chemical profiles were analyzed using gas chromatography-mass spectrometry (GC-MS). The in vitro antifungal activity of each extract was evaluated at different concentrations (500, 1000, 2000, 4000, and 8000 mg L-1) against phytopathogens such as Monilinia fructicola, Colletotrichum gloeosporioides, Fusarium oxysporum, and Sclerotinia sclerotiorum. Based on their chemical profiles, 14 compounds were identified in the hexanic extract, and 11 compounds were identified in the methanolic extract. These compounds included those with antifungal activity, such as Benzene; 1.3-bis(1.1-dimethylethyl)-; pentanoic acid; 5-hydroxy-, 2,4-di-1-butylphenyl esters; 1,2,4-Triazol-4-amine; and N-(2-thienylmethyl). The hexanic extract demonstrated fungistatic activity on the four fungi tested, while the methanolic extract exhibited fungicidal activity against C. gloeosporioides and F. oxysporum. The results of the Probit analysis showed variations in the sensitivity of phytopathogenic fungi to the treatments evaluated. In M. fructicola, the hexane extract presented an EC50 of 317,146 mg L-1 and an EC90 of 400,796 mg L-1. For C. gloeosporioides, the EC50 was 2676 mg L-1 and the EC90 was 888,177 mg L-1, while in F. oxysporum an EC50 of 34,274 mg L-1 and an EC90 of 1528 mg L-1 were estimated. In the case of S. sclerotiorum, an EC50 of 560 mg L-1 and an EC90 of 7776 mg L-1 were obtained. Finally, for the commercial fungicide Captan®, an EC50 of 1.19 mg L-1 and an EC90 of 1.67 mg L-1. These results suggest that extracts from A. mexicana could provide a natural alternative for the control of phytopathogenic fungi.
{"title":"Extracts of <i>Argemone mexicana</i> L. Contain Antifungal Compounds for the In Vitro Control of <i>Monilinia fructicola</i>, <i>Colletotrichum gloeosporioides, Fusarium oxysporum,</i> and <i>Sclerotinia sclerotiorum</i>: Preliminary Evidence for Field Application.","authors":"Iridiam Hernández-Soto, Antonio Juárez-Maldonado, Alfredo Madariaga-Navarrete, Ashutosh Sharma, Antonio de Jesus Cenobio-Galindo, Jose Manuel Pinedo-Espinoza, Aracely Hernández-Pérez, Alma Delia Hernández-Fuentes","doi":"10.3390/biotech14040082","DOIUrl":"10.3390/biotech14040082","url":null,"abstract":"<p><p><i>Argemone mexicana</i> L. is considered a weed; however, it contains secondary metabolites that can control phytopathogenic fungi in vitro, with the potential to adapt its effectiveness in the field. In the present study, leaf extracts of <i>A. mexicana</i> (hexane and methanol) were prepared, and their chemical profiles were analyzed using gas chromatography-mass spectrometry (GC-MS). The in vitro antifungal activity of each extract was evaluated at different concentrations (500, 1000, 2000, 4000, and 8000 mg L<sup>-1</sup>) against phytopathogens such as <i>Monilinia fructicola, Colletotrichum gloeosporioides, Fusarium oxysporum</i>, and <i>Sclerotinia sclerotiorum</i>. Based on their chemical profiles, 14 compounds were identified in the hexanic extract, and 11 compounds were identified in the methanolic extract. These compounds included those with antifungal activity, such as Benzene; 1.3-bis(1.1-dimethylethyl)-; pentanoic acid; 5-hydroxy-, 2,4-di-1-butylphenyl esters; 1,2,4-Triazol-4-amine; and N-(2-thienylmethyl). The hexanic extract demonstrated fungistatic activity on the four fungi tested, while the methanolic extract exhibited fungicidal activity against <i>C. gloeosporioides</i> and <i>F. oxysporum</i>. The results of the Probit analysis showed variations in the sensitivity of phytopathogenic fungi to the treatments evaluated. In <i>M. fructicola</i>, the hexane extract presented an EC<sub>50</sub> of 317,146 mg L<sup>-1</sup> and an EC<sub>90</sub> of 400,796 mg L<sup>-1</sup>. For <i>C. gloeosporioides</i>, the EC<sub>50</sub> was 2676 mg L<sup>-1</sup> and the EC<sub>90</sub> was 888,177 mg L<sup>-1</sup>, while in <i>F. oxysporum</i> an EC<sub>50</sub> of 34,274 mg L<sup>-1</sup> and an EC<sub>90</sub> of 1528 mg L<sup>-1</sup> were estimated. In the case of <i>S. sclerotiorum</i>, an EC<sub>50</sub> of 560 mg L<sup>-1</sup> and an EC<sub>90</sub> of 7776 mg L<sup>-1</sup> were obtained. Finally, for the commercial fungicide Captan<sup>®</sup>, an EC<sub>50</sub> of 1.19 mg L<sup>-1</sup> and an EC<sub>90</sub> of 1.67 mg L<sup>-1</sup>. These results suggest that extracts from <i>A. mexicana</i> could provide a natural alternative for the control of phytopathogenic fungi.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 4","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12641894/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145589251","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Newborn mice (up to 6 d after birth) are suitable for genetic manipulations, such as facial vein-mediated injection, owing to their hairless and thin skin. Their small body volumes also facilitate the rapid dissemination of injected solutions, supporting gene engineering-related experiments. However, anesthesia in newborns is challenging because of the potential risks associated with anesthetic agents. Isoflurane inhalation anesthesia is an option, although its effects on brain development remain under investigation. In this study, we established a reproducible protocol for delivering nucleic acids to juvenile mouse testes using a simple isoflurane-based anesthetic system prepared from common laboratory equipment. Using this system, nucleic acids were successfully delivered to juvenile mouse testes via intra-testicular injection, followed by in vivo electroporation. The present isoflurane-based method achieved >90% postoperative survival with normal maternal nursing observations. Gene delivery resulted in limited transfection of seminiferous tubules but efficient interstitial Leydig cell transfection. Thus, gene engineering in somatic and germ cells in neonatal mice will be facilitated using the anesthetic protocol established in this study.
{"title":"A Simple and Safe Protocol for Intra-Testicular Gene Delivery in Neonatal Mice Using a Convenient Isoflurane-Based Anesthesia System.","authors":"Kazunori Morohoshi, Miho Ohba, Masahiro Sato, Shingo Nakamura","doi":"10.3390/biotech14040081","DOIUrl":"10.3390/biotech14040081","url":null,"abstract":"<p><p>Newborn mice (up to 6 d after birth) are suitable for genetic manipulations, such as facial vein-mediated injection, owing to their hairless and thin skin. Their small body volumes also facilitate the rapid dissemination of injected solutions, supporting gene engineering-related experiments. However, anesthesia in newborns is challenging because of the potential risks associated with anesthetic agents. Isoflurane inhalation anesthesia is an option, although its effects on brain development remain under investigation. In this study, we established a reproducible protocol for delivering nucleic acids to juvenile mouse testes using a simple isoflurane-based anesthetic system prepared from common laboratory equipment. Using this system, nucleic acids were successfully delivered to juvenile mouse testes via intra-testicular injection, followed by in vivo electroporation. The present isoflurane-based method achieved >90% postoperative survival with normal maternal nursing observations. Gene delivery resulted in limited transfection of seminiferous tubules but efficient interstitial Leydig cell transfection. Thus, gene engineering in somatic and germ cells in neonatal mice will be facilitated using the anesthetic protocol established in this study.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 4","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12641744/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145589169","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
María Isabel Iñiguez-Luna, Jorge David Cadena-Zamudio, Marco A Ramírez-Mosqueda, José Luis Aguirre-Noyola, Daniel Alejandro Cadena-Zamudio, Jorge Cadena-Iñiguez, Alma Armenta-Medina
Molecular docking has emerged as a pivotal computational approach in agri-food research, offering a rapid and targeted means to discover bioactive molecules for crop protection and food safety. Its ability to predict and visualize interactions between natural or synthetic compounds and specific biological targets provides valuable opportunities to address urgent agricultural challenges, including climate change and the rise in resistant crop pathogens. By enabling the in silico screening of diverse chemical entities, this technique facilitates the identification of molecules with antimicrobial and antifungal properties, specifically designed to interact with critical enzymatic pathways in plant pathogens. Recent advancements, such as the integration of molecular dynamics simulations and artificial intelligence-enhanced scoring functions, have significantly improved docking accuracy by addressing limitations like protein flexibility and solvent effects. These technological improvements have accelerated the discovery of eco-friendly biopesticides and multifunctional nutraceutical agents. Promising developments include nanoparticle-based delivery systems that enhance the stability and efficacy of bioactive molecules. Despite its potential, molecular docking still faces challenges related to incomplete protein structures, variability in scoring algorithms, and limited experimental validation in agricultural contexts. This work highlights these limitations while outlining current trends and future prospects to guide its effective application in agri-food biotechnology.
{"title":"Molecular Docking as a Key Driver of Biocontrol for Agri-Food Security.","authors":"María Isabel Iñiguez-Luna, Jorge David Cadena-Zamudio, Marco A Ramírez-Mosqueda, José Luis Aguirre-Noyola, Daniel Alejandro Cadena-Zamudio, Jorge Cadena-Iñiguez, Alma Armenta-Medina","doi":"10.3390/biotech14040080","DOIUrl":"10.3390/biotech14040080","url":null,"abstract":"<p><p>Molecular docking has emerged as a pivotal computational approach in agri-food research, offering a rapid and targeted means to discover bioactive molecules for crop protection and food safety. Its ability to predict and visualize interactions between natural or synthetic compounds and specific biological targets provides valuable opportunities to address urgent agricultural challenges, including climate change and the rise in resistant crop pathogens. By enabling the in silico screening of diverse chemical entities, this technique facilitates the identification of molecules with antimicrobial and antifungal properties, specifically designed to interact with critical enzymatic pathways in plant pathogens. Recent advancements, such as the integration of molecular dynamics simulations and artificial intelligence-enhanced scoring functions, have significantly improved docking accuracy by addressing limitations like protein flexibility and solvent effects. These technological improvements have accelerated the discovery of eco-friendly biopesticides and multifunctional nutraceutical agents. Promising developments include nanoparticle-based delivery systems that enhance the stability and efficacy of bioactive molecules. Despite its potential, molecular docking still faces challenges related to incomplete protein structures, variability in scoring algorithms, and limited experimental validation in agricultural contexts. This work highlights these limitations while outlining current trends and future prospects to guide its effective application in agri-food biotechnology.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 4","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12550905/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145355807","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Gloria Sciuto, Nunziatina Russo, Cinzia L Randazzo, Cinzia Caggia
The dairy sector produces considerable amounts of nutrient-rich effluents, which are frequently undervalued as simple by-products or waste. In particular, Second Cheese Whey (SCW), also known as scotta, exhausted whey, or deproteinized whey, represents the liquid fraction from ricotta cheese production. Despite its abundance and high organic and saline content, SCW is often improperly discharged into terrestrial and aquatic ecosystems, causing both environmental impact and resource waste. The available purification methods are expensive for dairy companies, and, at best, SCW is reused as feed or fertilizer. In recent years, increasing awareness of sustainability and circular economy principles has increased interest in the valorization of SCW. Biological treatment of SCW using microalgae represents an attractive strategy, as it simultaneously reduces the organic load and converts waste into algal biomass. This biomass can be further valorized as a source of proteins, pigments, and bioactive compounds with industrial relevance, supporting applications in food, nutraceuticals, biofuels, and cosmetics. This review, starting from analyzing the characteristics, production volumes, and environmental issues associated with SCW, focused on the potential of microalgae application for their valorization. In addition, the broader regulatory and sustainability aspects related to biomass utilization and treated SCW are considered, highlighting both the promises and limitations of microalgae-based strategies by integrating technological prospects with policy considerations.
{"title":"Valorization of Second Cheese Whey Through Microalgae-Based Treatments: Advantages, Limits, and Opportunities.","authors":"Gloria Sciuto, Nunziatina Russo, Cinzia L Randazzo, Cinzia Caggia","doi":"10.3390/biotech14040079","DOIUrl":"10.3390/biotech14040079","url":null,"abstract":"<p><p>The dairy sector produces considerable amounts of nutrient-rich effluents, which are frequently undervalued as simple by-products or waste. In particular, Second Cheese Whey (SCW), also known as scotta, exhausted whey, or deproteinized whey, represents the liquid fraction from ricotta cheese production. Despite its abundance and high organic and saline content, SCW is often improperly discharged into terrestrial and aquatic ecosystems, causing both environmental impact and resource waste. The available purification methods are expensive for dairy companies, and, at best, SCW is reused as feed or fertilizer. In recent years, increasing awareness of sustainability and circular economy principles has increased interest in the valorization of SCW. Biological treatment of SCW using microalgae represents an attractive strategy, as it simultaneously reduces the organic load and converts waste into algal biomass. This biomass can be further valorized as a source of proteins, pigments, and bioactive compounds with industrial relevance, supporting applications in food, nutraceuticals, biofuels, and cosmetics. This review, starting from analyzing the characteristics, production volumes, and environmental issues associated with SCW, focused on the potential of microalgae application for their valorization. In addition, the broader regulatory and sustainability aspects related to biomass utilization and treated SCW are considered, highlighting both the promises and limitations of microalgae-based strategies by integrating technological prospects with policy considerations.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 4","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12550978/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145356081","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The detection of allergens is essential for ensuring food safety, protecting public health, and providing accurate information to consumers. Wheat (Triticum aestivum L.) and maize (Zea mays L.) are recognized as important food allergens. In this study, novel PCR methods were developed for the reliable detection of wheat and maize allergens, including wheat high-molecular-weight glutenin subunit (HMW-GS) and low-molecular-weight glutenin subunit (LMW-GS), as well as three maize allergens, namely, Zea m 14, Zea m 8, and zein. Wheat and maize genomic DNA, as well as allergen genes, were examined during 60 min of baking at 180 °C and 220 °C. Agarose gel electrophoresis revealed degradation of genomic DNA and amplified PCR fragments in correlation with increasing baking temperature and time. For each target gene, the best primers were identified that could detect HMW-GS and LMW-GS genes in wheat samples and Zea m 14, Zea m 8, and zein genes in maize samples after baking at 220 °C for 60 min and 40 min, respectively. The results indicate that these PCR methods can be used for the reliable and sensitive detection of wheat and maize allergens in processed foods.
检测过敏原对于确保食品安全、保护公众健康和向消费者提供准确信息至关重要。小麦(Triticum aestivum L.)和玉米(Zea mays L.)是公认的重要食物过敏原。本研究建立了小麦和玉米高分子量谷蛋白亚基(HMW-GS)和低分子量谷蛋白亚基(LMW-GS)以及玉米3种过敏原(Zea m 14、Zea m 8和zein)的可靠检测方法。在180°C和220°C烘烤60分钟期间,检测小麦和玉米基因组DNA以及过敏原基因。琼脂糖凝胶电泳显示基因组DNA的降解和扩增的PCR片段与烘烤温度和时间的增加有关。针对每个目标基因,分别在220℃烘烤60 min和40 min后,鉴定出能检测小麦样品中HMW-GS和LMW-GS基因以及玉米样品中Zea m 14、Zea m 8和zein基因的最佳引物。结果表明,该方法可可靠、灵敏地检测加工食品中的小麦和玉米过敏原。
{"title":"Development of PCR Methods for Detecting Wheat and Maize Allergens in Food.","authors":"Tata Ninidze, Tamar Koberidze, Kakha Bitskinashvili, Tamara Kutateladze, Boris Vishnepolsky, Nelly Datukishvili","doi":"10.3390/biotech14040078","DOIUrl":"10.3390/biotech14040078","url":null,"abstract":"<p><p>The detection of allergens is essential for ensuring food safety, protecting public health, and providing accurate information to consumers. Wheat (<i>Triticum aestivum</i> L.) and maize (<i>Zea mays</i> L.) are recognized as important food allergens. In this study, novel PCR methods were developed for the reliable detection of wheat and maize allergens, including wheat high-molecular-weight glutenin subunit (HMW-GS) and low-molecular-weight glutenin subunit (LMW-GS), as well as three maize allergens, namely, Zea m 14, Zea m 8, and zein. Wheat and maize genomic DNA, as well as allergen genes, were examined during 60 min of baking at 180 °C and 220 °C. Agarose gel electrophoresis revealed degradation of genomic DNA and amplified PCR fragments in correlation with increasing baking temperature and time. For each target gene, the best primers were identified that could detect HMW-GS and LMW-GS genes in wheat samples and Zea m 14, Zea m 8, and zein genes in maize samples after baking at 220 °C for 60 min and 40 min, respectively. The results indicate that these PCR methods can be used for the reliable and sensitive detection of wheat and maize allergens in processed foods.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"14 4","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12551070/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145356272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}