The study of protein-protein interactions (PPIs) is of significant importance for elucidating biological processes, clarifying pathological mechanisms, and promoting drug development. In this study, we proposed a method to predict PPIs based on protein sequence and gene sequence information, combined with convolutional neural networks (CNNs). First, we extracted three types of features from protein sequence: global physicochemical properties features of the protein sequence, local same type of amino acid position variation features, and protein evolutionary conservation features; simultaneously, we extracted single nucleotide frequency and positional features, dinucleotide frequency features, and trinucleotide frequency features from the corresponding gene sequence. During the feature extraction process, we employed the amphiphilic pseudo amino acid composition (APAAC) method to extract the global hydrophobicity and hydrophilicity features of the protein sequence; we defined a new mathematical descriptor-θ interval deviation product factor-to extract protein evolutionary conservation features from Position Specific Scoring Matrix (PSSM); we also defined a mapping function to map all nucleotides in the gene sequence onto a unit circle, and then extracted nucleotide positional features from the mapped points. Second, based on extracted features, we constructed a 36 × 32 sample feature grayscale map to represent a protein pair sample. Finally, we developed a CNN model to predict PPIs. Our method achieved superior results on four species test sets: an accuracy of 99.28% on the Saccharomyces cerevisiae dataset, 98.15% on the Drosophila melanogaster dataset, 98.62% on the Homo sapiens dataset, and 96.84% on the Mus musculus dataset, outperforming existing computational methods. Furthermore, we extended the application of this method to the prediction of protein-protein interaction networks and non-interaction networks, and also achieved promising results.
{"title":"Predicting Protein-Protein Interactions by Convolutional Neural Network Model.","authors":"Shuaibo Shi, Ting Xiong, Dong Wang, Lingling Wei, Lin Li, Zhixin Li, Yanfen Lyu","doi":"10.3390/biotech15010020","DOIUrl":"10.3390/biotech15010020","url":null,"abstract":"<p><p>The study of protein-protein interactions (PPIs) is of significant importance for elucidating biological processes, clarifying pathological mechanisms, and promoting drug development. In this study, we proposed a method to predict PPIs based on protein sequence and gene sequence information, combined with convolutional neural networks (CNNs). First, we extracted three types of features from protein sequence: global physicochemical properties features of the protein sequence, local same type of amino acid position variation features, and protein evolutionary conservation features; simultaneously, we extracted single nucleotide frequency and positional features, dinucleotide frequency features, and trinucleotide frequency features from the corresponding gene sequence. During the feature extraction process, we employed the amphiphilic pseudo amino acid composition (APAAC) method to extract the global hydrophobicity and hydrophilicity features of the protein sequence; we defined a new mathematical descriptor-θ interval deviation product factor-to extract protein evolutionary conservation features from Position Specific Scoring Matrix (PSSM); we also defined a mapping function to map all nucleotides in the gene sequence onto a unit circle, and then extracted nucleotide positional features from the mapped points. Second, based on extracted features, we constructed a 36 × 32 sample feature grayscale map to represent a protein pair sample. Finally, we developed a CNN model to predict PPIs. Our method achieved superior results on four species test sets: an accuracy of 99.28% on the <i>Saccharomyces cerevisiae</i> dataset, 98.15% on the <i>Drosophila melanogaster</i> dataset, 98.62% on the <i>Homo sapiens</i> dataset, and 96.84% on the <i>Mus musculus</i> dataset, outperforming existing computational methods. Furthermore, we extended the application of this method to the prediction of protein-protein interaction networks and non-interaction networks, and also achieved promising results.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"15 1","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12938199/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146259578","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}
Christiana Bitrus, Ademola Hammed, Tawakalt Ayodele, Niloy Chandra Sarker
Background/objectives: The effects of thermal drying on the viability of beneficial microorganisms immobilized in biochar, as well as on biochar nutrient retention, remain insufficiently understood. This study aimed to evaluate how drying temperature influences the survival of hyper-ammonia-producing bacteria (HAB) immobilized on pine wood biochar and to assess the impact of subsequent storage on bacterial recovery and nutrient stability.
Methods: Biochar was inoculated with HAB and subjected to drying at temperatures ranging from 40 to 60 °C. Following drying, samples were characterized and stored for 30 days. Microbial revival was assessed through reculturing, while changes in surface functional groups were analyzed using FTIR spectroscopy. Nutrient retention, particularly nitrogen content, was also evaluated.
Results: Higher drying temperatures resulted in reduced immediate microbial revival during reculturing. However, samples exhibiting limited immediate recovery demonstrated enhanced revival after the 30-day storage period. FTIR analysis revealed that drying temperature modified the availability of surface functional groups associated with microbial attachment and activity. Nutrient analysis indicated only minor reductions in nitrogen retention in biochar dried at temperatures above 55 °C.
Conclusions: Drying temperature significantly affects both the short-term survival and post-storage recovery of beneficial microorganisms immobilized in biochar. While elevated temperatures may initially suppress microbial activity, recovery potential during storage remains substantial. Optimizing drying conditions is therefore essential to balance microbial viability with nutrient retention in biochar-based formulations.
{"title":"Post-Inoculation Drying and Storage Effects on HAB Viability and Nutrient Retention in Biochar.","authors":"Christiana Bitrus, Ademola Hammed, Tawakalt Ayodele, Niloy Chandra Sarker","doi":"10.3390/biotech15010019","DOIUrl":"10.3390/biotech15010019","url":null,"abstract":"<p><strong>Background/objectives: </strong>The effects of thermal drying on the viability of beneficial microorganisms immobilized in biochar, as well as on biochar nutrient retention, remain insufficiently understood. This study aimed to evaluate how drying temperature influences the survival of hyper-ammonia-producing bacteria (HAB) immobilized on pine wood biochar and to assess the impact of subsequent storage on bacterial recovery and nutrient stability.</p><p><strong>Methods: </strong>Biochar was inoculated with HAB and subjected to drying at temperatures ranging from 40 to 60 °C. Following drying, samples were characterized and stored for 30 days. Microbial revival was assessed through reculturing, while changes in surface functional groups were analyzed using FTIR spectroscopy. Nutrient retention, particularly nitrogen content, was also evaluated.</p><p><strong>Results: </strong>Higher drying temperatures resulted in reduced immediate microbial revival during reculturing. However, samples exhibiting limited immediate recovery demonstrated enhanced revival after the 30-day storage period. FTIR analysis revealed that drying temperature modified the availability of surface functional groups associated with microbial attachment and activity. Nutrient analysis indicated only minor reductions in nitrogen retention in biochar dried at temperatures above 55 °C.</p><p><strong>Conclusions: </strong>Drying temperature significantly affects both the short-term survival and post-storage recovery of beneficial microorganisms immobilized in biochar. While elevated temperatures may initially suppress microbial activity, recovery potential during storage remains substantial. Optimizing drying conditions is therefore essential to balance microbial viability with nutrient retention in biochar-based formulations.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"15 1","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12922006/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146259520","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}
Angelo Michilli, Cristian Bassi, Farzaneh Moshiri, Bruno De Siena, Rosaria Marinaro, Elisa Callegari, Massimo Negrini, Silvia Sabbioni
Despite the advent of immune checkpoint inhibitor-based regimens, sorafenib remains an important therapeutic option for patients with advanced hepatocellular carcinoma (HCC) who are ineligible for immunotherapy. However, its clinical efficacy is limited by the emergence of drug resistance, whose underlying molecular mechanisms remain incompletely understood. To investigate these mechanisms, we established a murine model of acquired sorafenib resistance and performed comparative RNA sequencing of sorafenib-sensitive versus -resistant Hep55.1C hepatoma cells. Transcriptomic profiling revealed a distinct resistance-associated signature comprising 1264 significantly deregulated genes (adjusted p < 0.03, fold change > 3.0). Pathway analysis and Gene Set Enrichment Analyses (GSEA) indicated a coordinated downregulation of metabolic and intercellular signaling pathways, accompanied by marked upregulation of redox-regulatory, mitochondrial and cellular stress-response programs. Genes transcriptionally regulated by nuclear factor erythroid 2-related factor 2 (NRF2) including Gpx4, Txn1, Txnrd1, Hmox1, Fth1, Taldo1, Phgdh, and MafG, involved in antioxidant defense, ferroptosis suppression and metabolic rewiring, were all upregulated in resistant cells. Pharmacological inhibition of NRF2 activity using brusatol restored sensitivity to sorafenib, functionally implicating NRF2-dependent pathways in the maintenance of the resistant phenotype. These findings demonstrate that acquired sorafenib resistance in HCC is associated with a stable NRF2-driven transcriptional and metabolic reprogramming that enhances antioxidant capacity, suppresses ferroptosis and promotes tumor cell survival. Targeting NRF2-regulated redox metabolism may therefore represent a promising strategy to overcome therapeutic resistance in HCC.
{"title":"Transcriptomic Analysis Reveals an NRF2-Mediated Redox and Metabolic Reprogramming in Sorafenib-Resistant Hepatocellular Carcinoma Cells.","authors":"Angelo Michilli, Cristian Bassi, Farzaneh Moshiri, Bruno De Siena, Rosaria Marinaro, Elisa Callegari, Massimo Negrini, Silvia Sabbioni","doi":"10.3390/biotech15010018","DOIUrl":"10.3390/biotech15010018","url":null,"abstract":"<p><p>Despite the advent of immune checkpoint inhibitor-based regimens, sorafenib remains an important therapeutic option for patients with advanced hepatocellular carcinoma (HCC) who are ineligible for immunotherapy. However, its clinical efficacy is limited by the emergence of drug resistance, whose underlying molecular mechanisms remain incompletely understood. To investigate these mechanisms, we established a murine model of acquired sorafenib resistance and performed comparative RNA sequencing of sorafenib-sensitive versus -resistant Hep55.1C hepatoma cells. Transcriptomic profiling revealed a distinct resistance-associated signature comprising 1264 significantly deregulated genes (adjusted <i>p</i> < 0.03, fold change > 3.0). Pathway analysis and Gene Set Enrichment Analyses (GSEA) indicated a coordinated downregulation of metabolic and intercellular signaling pathways, accompanied by marked upregulation of redox-regulatory, mitochondrial and cellular stress-response programs. Genes transcriptionally regulated by nuclear factor erythroid 2-related factor 2 (NRF2) including Gpx4, Txn1, Txnrd1, Hmox1, Fth1, Taldo1, Phgdh, and MafG, involved in antioxidant defense, ferroptosis suppression and metabolic rewiring, were all upregulated in resistant cells. Pharmacological inhibition of NRF2 activity using brusatol restored sensitivity to sorafenib, functionally implicating NRF2-dependent pathways in the maintenance of the resistant phenotype. These findings demonstrate that acquired sorafenib resistance in HCC is associated with a stable NRF2-driven transcriptional and metabolic reprogramming that enhances antioxidant capacity, suppresses ferroptosis and promotes tumor cell survival. Targeting NRF2-regulated redox metabolism may therefore represent a promising strategy to overcome therapeutic resistance in HCC.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"15 1","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921911/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146259581","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}
Acidogenic fermentation is a promising biotechnology for converting organic wastes into carboxylic acid (CA), which has significant commercial value and diverse applications in the food, chemical, pharmaceutical, and cosmetic industries. However, major challenges such as limited substrate hydrolysis and lower CA production hinder further development of this biotechnology towards full-scale implementation. This review provides a comprehensive overview of the current status of acidogenic fermentation, focusing on substrate composition, inoculum, and reactor design, along with potential strategies to overcome reactor-specific limitations and enhance CA production. It was found that the substrate composition, particularly its carbohydrate, protein, and lipid contents, strongly influences both CA production and yield. Specifically, carbohydrate-rich substrates yield higher CA production compared to protein- and lipid-rich substrates. These substrates have been investigated in different reactor configurations for CA production. Among them, the leachate bed reactor and anaerobic membrane bioreactor have demonstrated superior performance, achieving higher CA production with acetic and butyric acids as the dominant CA composition. These reactors are generally operated using three types of inocula: aerobic and anaerobic inoculum, enriched inoculum, and rumen microorganisms. Interestingly, rumen microorganisms are effective in degrading complex substrates, whereas enriched inoculum accelerates hydrolysis and acidogenesis processes within a shorter fermentation time. The findings presented herein will provide valuable information for addressing the challenges associated with acidogenic fermentation and lay the foundation for future research aimed at upscaling this biotechnology to a commercial scale.
{"title":"Carboxylic Acid Production from Organic Waste: Integrating Substrate Composition, Reactor Configuration, Inoculum, and Future Perspectives.","authors":"Ajay Thapa, Shiyu Fu, Joseph Sebastian, Onita Basu, Farah Hosseinian, Utsav Sharma, Dayanand Sharma, Abid Hussain","doi":"10.3390/biotech15010016","DOIUrl":"10.3390/biotech15010016","url":null,"abstract":"<p><p>Acidogenic fermentation is a promising biotechnology for converting organic wastes into carboxylic acid (CA), which has significant commercial value and diverse applications in the food, chemical, pharmaceutical, and cosmetic industries. However, major challenges such as limited substrate hydrolysis and lower CA production hinder further development of this biotechnology towards full-scale implementation. This review provides a comprehensive overview of the current status of acidogenic fermentation, focusing on substrate composition, inoculum, and reactor design, along with potential strategies to overcome reactor-specific limitations and enhance CA production. It was found that the substrate composition, particularly its carbohydrate, protein, and lipid contents, strongly influences both CA production and yield. Specifically, carbohydrate-rich substrates yield higher CA production compared to protein- and lipid-rich substrates. These substrates have been investigated in different reactor configurations for CA production. Among them, the leachate bed reactor and anaerobic membrane bioreactor have demonstrated superior performance, achieving higher CA production with acetic and butyric acids as the dominant CA composition. These reactors are generally operated using three types of inocula: aerobic and anaerobic inoculum, enriched inoculum, and rumen microorganisms. Interestingly, rumen microorganisms are effective in degrading complex substrates, whereas enriched inoculum accelerates hydrolysis and acidogenesis processes within a shorter fermentation time. The findings presented herein will provide valuable information for addressing the challenges associated with acidogenic fermentation and lay the foundation for future research aimed at upscaling this biotechnology to a commercial scale.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"15 1","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12922049/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146259429","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}
Species of the genus Phytophthora are among the most detrimental plant pathogens globally, representing a significant threat to global agriculture, horticulture, and forestry. These zoosporic oomycetes have historically caused devastating outbreaks, including, just to mention a few, late blight of potato in Ireland; jarrah dieback of eucalyptus in Western Australia; ink disease of chestnut in Europe; sudden oak death and sudden larch death of coast live oak and tanoak in the Western US, and of Japanese larch in the UK. The environmental and ecological impacts of the diseases they cause result in significant economic costs that often have social repercussions. With the acceleration of globalization, enhancing the movement of plant material, in particular with the global live plant trade, the spread of Phytophthora to new, uncontaminated territories has intensified. Nurseries play a key role in the movement of these pathogens, the trade of contaminated stocks representing their major dissemination route. However valuable, conventional detection techniques, including baiting and direct isolation, are too slow and labour-intensive to meet current diagnostic requirements, particularly given the huge volumes of plants traded globally. This problem becomes even more acute when large volumes of potentially infectious plant material need to be processed in a short time frame, as it is often necessary to provide accurate and timely responses to interested parties. Early and precise detection is thus vital to avert outbreaks and mitigate long-term consequences. This review evaluates and contrasts the efficacy of novel detection methods against traditional approaches, emphasizing their significance in managing the escalating threat posed by Phytophthora spp. worldwide. Despite technological advances, critical challenges remain that limit the reliability and large-scale adoption of new diagnostic methods. Research still needs to bridge the gap between the laboratory and the field in terms of accuracy, sensitivity and diagnostic costs. Recent innovations focus on sensor technology and point-of-care (POC) devices for faster, more sensitive, and low-cost specific detection of Phytophthora spp. in plant matrices, water and soil. Enhancing diagnostic capabilities through these tools is crucial for protecting agricultural productivity, local economies, and natural ecosystems.
{"title":"Catching the Elusive <i>Phytophthora</i>: A Review of Methods and Applications for Pathogen Detection and Identification Across Agricultural, Horticultural, Forestry and Ornamental Settings.","authors":"Viola Papini, Alessandra Benigno, Domenico Rizzo, Salvatore Moricca","doi":"10.3390/biotech15010017","DOIUrl":"10.3390/biotech15010017","url":null,"abstract":"<p><p>Species of the genus <i>Phytophthora</i> are among the most detrimental plant pathogens globally, representing a significant threat to global agriculture, horticulture, and forestry. These zoosporic oomycetes have historically caused devastating outbreaks, including, just to mention a few, late blight of potato in Ireland; jarrah dieback of eucalyptus in Western Australia; ink disease of chestnut in Europe; sudden oak death and sudden larch death of coast live oak and tanoak in the Western US, and of Japanese larch in the UK. The environmental and ecological impacts of the diseases they cause result in significant economic costs that often have social repercussions. With the acceleration of globalization, enhancing the movement of plant material, in particular with the global live plant trade, the spread of <i>Phytophthora</i> to new, uncontaminated territories has intensified. Nurseries play a key role in the movement of these pathogens, the trade of contaminated stocks representing their major dissemination route. However valuable, conventional detection techniques, including baiting and direct isolation, are too slow and labour-intensive to meet current diagnostic requirements, particularly given the huge volumes of plants traded globally. This problem becomes even more acute when large volumes of potentially infectious plant material need to be processed in a short time frame, as it is often necessary to provide accurate and timely responses to interested parties. Early and precise detection is thus vital to avert outbreaks and mitigate long-term consequences. This review evaluates and contrasts the efficacy of novel detection methods against traditional approaches, emphasizing their significance in managing the escalating threat posed by <i>Phytophthora</i> spp. worldwide. Despite technological advances, critical challenges remain that limit the reliability and large-scale adoption of new diagnostic methods. Research still needs to bridge the gap between the laboratory and the field in terms of accuracy, sensitivity and diagnostic costs. Recent innovations focus on sensor technology and point-of-care (POC) devices for faster, more sensitive, and low-cost specific detection of <i>Phytophthora</i> spp. in plant matrices, water and soil. Enhancing diagnostic capabilities through these tools is crucial for protecting agricultural productivity, local economies, and natural ecosystems.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"15 1","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12922092/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146259451","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}
Human embryonic kidney (HEK293) cells are a widespread choice for recombinant protein expression. To optimise yields, the hydrolysate Tryptone N1 (TN1) is commonly added post-transfection. TN1 is obtained by controlled enzymatic digestion of casein. As an animal by-product, TN1 faces stricter regulations during cross-country shipments than plant-based products. This raises the question of whether plant-derived peptides are a suitable alternative to TN1. Using polyethyleneimine (PEI) as a cationic polymer, we transfected HEK293-6E cells grown in suspension in serum-free medium and divided the transfectants into four groups (each in triplicate). Two plant-based hydrolysates each derived from pea and broad bean were compared with TN1 and a no-hydrolysate control group. We monitored the cultures for total cell numbers and viability at days 1, 4, and 5 post-transfection. Both plant-based hydrolysates and TN1 showed similar live cell percentages, in contrast to the no-hydrolysate control, which showed lower viability. Five days post-transfection, the expressed His-tagged protein, a tegumental antigen from the eukaryotic parasite Echinococcus granulosus, was retrieved from the serum-free culture supernatant, and the expressed recombinant protein was quantified. The linear ranges for the protein load on the stain-free blot and for the use of the fluorescent anti-His-Tag Alexa488 antibody were determined. Using these parameters, stain-free Western blotting and total protein normalization were performed. The plant-derived pea and broad bean hydrolysates reproducibly resulted in similar expression levels as animal-derived TN1; all three hydrolysates were better than no hydrolysate. We conclude that plant-derived hydrolysates are a suitable, more sustainable replacement for TN1.
人胚胎肾(HEK293)细胞是重组蛋白表达的广泛选择。为了优化产量,通常在转染后添加水解的Tryptone N1 (TN1)。TN1是通过控制酪蛋白的酶消化得到的。作为动物副产品,TN1在跨国运输中面临着比植物产品更严格的监管。这就提出了一个问题,即植物来源的肽是否是TN1的合适替代品。我们以聚乙烯亚胺(PEI)为阳离子聚合物,转染悬浮生长于无血清培养基中的HEK293-6E细胞,并将转染物分为四组(每组三份)。从豌豆和蚕豆中提取的两种植物性水解物与TN1和无水解物对照组进行了比较。我们在转染后第1,4,5天监测培养物的总细胞数和活力。植物基水解物和TN1都显示出相似的活细胞百分比,而非水解物的对照组则显示出较低的活力。转染5天后,从无血清培养上清中提取真核寄生虫细粒棘球蚴(Echinococcus granulosus)的被膜抗原表达his标记蛋白,并对表达的重组蛋白进行定量分析。确定了无染色印迹和荧光抗his - tag Alexa488抗体上蛋白负荷的线性范围。利用这些参数进行无染色Western blotting和总蛋白归一化。植物源性豌豆和蚕豆水解物可重复性地产生与动物源性TN1相似的表达水平;三种水解物均优于无水解物。我们得出结论,植物衍生的水解物是一种合适的、更可持续的TN1替代品。
{"title":"Plant-Derived Hydrolysates Are a Suitable Replacement for Tryptone N1 in Recombinant Protein Expression Using Human Embryonic Kidney (HEK293-6E) Cells.","authors":"Shafqat Shabir, Md Shahadat Hossain, Lucie Egly, Gizem Yalkin, Franco H Falcone","doi":"10.3390/biotech15010014","DOIUrl":"10.3390/biotech15010014","url":null,"abstract":"<p><p>Human embryonic kidney (HEK293) cells are a widespread choice for recombinant protein expression. To optimise yields, the hydrolysate Tryptone N1 (TN1) is commonly added post-transfection. TN1 is obtained by controlled enzymatic digestion of casein. As an animal by-product, TN1 faces stricter regulations during cross-country shipments than plant-based products. This raises the question of whether plant-derived peptides are a suitable alternative to TN1. Using polyethyleneimine (PEI) as a cationic polymer, we transfected HEK293-6E cells grown in suspension in serum-free medium and divided the transfectants into four groups (each in triplicate). Two plant-based hydrolysates each derived from pea and broad bean were compared with TN1 and a no-hydrolysate control group. We monitored the cultures for total cell numbers and viability at days 1, 4, and 5 post-transfection. Both plant-based hydrolysates and TN1 showed similar live cell percentages, in contrast to the no-hydrolysate control, which showed lower viability. Five days post-transfection, the expressed His-tagged protein, a tegumental antigen from the eukaryotic parasite <i>Echinococcus granulosus</i>, was retrieved from the serum-free culture supernatant, and the expressed recombinant protein was quantified. The linear ranges for the protein load on the stain-free blot and for the use of the fluorescent anti-His-Tag Alexa<sub>488</sub> antibody were determined. Using these parameters, stain-free Western blotting and total protein normalization were performed. The plant-derived pea and broad bean hydrolysates reproducibly resulted in similar expression levels as animal-derived TN1; all three hydrolysates were better than no hydrolysate. We conclude that plant-derived hydrolysates are a suitable, more sustainable replacement for TN1.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"15 1","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12922121/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146259534","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}
Carotenoid-based pigmentation is crucial for the ornamental and commercial value of the cherry shrimp (Neocaridina denticulata sinensis). While several genes are known to influence carotenoid metabolism, the genetic basis for specific color strains remains largely unexplored. Here, we functionally characterized NinaB-like, a homolog of a carotenoid oxygenase, in cherry shrimp pigmentation. We employed qPCR to gain gene expression profiles, utilized RNAi technology to analysize the relation between its expression level and carotenoid accumulation, and performed GT-seq to identify genotypes of different color strains. Significant differential expression of NinaB-like was observed not only across distinct color strains but also during embryonic development of cherry shrimp (p < 0.05), peaking at the red strain and post-larval stage of cherry shrimp. RNA interference-mediated knockdown of NinaB-like resulted in a marked increase in red pigment deposition at the metanauplius and pre-zoea stages, confirming its role as a negative regulator of carotenoid accumulation. Importantly, we identified two tightly linked, non-synonymous SNPs (927C > A and 935A > C) within the NinaB-like coding region that exhibited a strong association with body color. Our study provides the first functional evidence that NinaB-like is a negative regulator of carotenoid degradation and a major genetic determinant for body color in cherry shrimp, providing new insights for genetic breeding and biological research.
{"title":"Functional Analysis of the <i>NinaB-like</i> Gene in Body Color Regulation of <i>Neocaridina denticulata sinensis</i>.","authors":"Haifan Li, Lili Zhang, Guodong Wang, Tanjun Zhao","doi":"10.3390/biotech15010015","DOIUrl":"10.3390/biotech15010015","url":null,"abstract":"<p><p>Carotenoid-based pigmentation is crucial for the ornamental and commercial value of the cherry shrimp (<i>Neocaridina denticulata sinensis</i>). While several genes are known to influence carotenoid metabolism, the genetic basis for specific color strains remains largely unexplored. Here, we functionally characterized <i>NinaB-like</i>, a homolog of a carotenoid oxygenase, in cherry shrimp pigmentation. We employed qPCR to gain gene expression profiles, utilized RNAi technology to analysize the relation between its expression level and carotenoid accumulation, and performed GT-seq to identify genotypes of different color strains. Significant differential expression of <i>NinaB-like</i> was observed not only across distinct color strains but also during embryonic development of cherry shrimp (<i>p</i> < 0.05), peaking at the red strain and post-larval stage of cherry shrimp. RNA interference-mediated knockdown of <i>NinaB-like</i> resulted in a marked increase in red pigment deposition at the metanauplius and pre-zoea stages, confirming its role as a negative regulator of carotenoid accumulation. Importantly, we identified two tightly linked, non-synonymous SNPs (927C > A and 935A > C) within the <i>NinaB-like</i> coding region that exhibited a strong association with body color. Our study provides the first functional evidence that <i>NinaB-like</i> is a negative regulator of carotenoid degradation and a major genetic determinant for body color in cherry shrimp, providing new insights for genetic breeding and biological research.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"15 1","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12922109/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146259506","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}
Phuong Duy Nguyen, Van Thi Pham, Ha Thanh Nguyen, Khoa Dang Dang, Tu Tuan Tran, Dai Lan Tran, Thanh Duc Nguyen, Thao Duc Le, Xuan Hoi Pham, Xuan Dang Tran, Quyen Le Cao
This study investigates the functional role of OsHSBP1, a heat shock factor-binding protein, in regulating abiotic stress tolerance in rice, with the aim of enhancing climate resilience in the elite indica cultivar Bacthom 7 (BT7). Using Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9 (CRISPR/Cas9) genome editing, we generated transgene-free homozygous knockout lines targeting OsHSBP1 and evaluated their physiological, biochemical, and agronomic responses under heat stress. Mutant lines exhibited markedly improved tolerance to both stresses, with survival rates reaching 43-46% under heat stress, compared to near-zero in wildtype plants. Enhanced tolerance was associated with significantly increased catalase and peroxidase activities and reduced oxidative damage, including lower malondialdehyde content and decreased superoxide accumulation. Despite these stress-related advantages, the knockout lines showed minimal differences in key agronomic traits under normal growing conditions, with comparable plant height, tillering ability, grain yield, and amylose content relative to the wildtype. These results demonstrate that OsHSBP1 functions as a negative regulator of abiotic stress tolerance in rice, and its knockout enhances resilience without compromising yield potential. The study highlights OsHSBP1 as a promising target for precision breeding of climate-resilient rice cultivars.
{"title":"CRISPR/Cas9-Mediated Knockout of <i>OsHSBP1</i> Confers Heat Tolerance to Bacthom 7 Elite Rice Cultivar.","authors":"Phuong Duy Nguyen, Van Thi Pham, Ha Thanh Nguyen, Khoa Dang Dang, Tu Tuan Tran, Dai Lan Tran, Thanh Duc Nguyen, Thao Duc Le, Xuan Hoi Pham, Xuan Dang Tran, Quyen Le Cao","doi":"10.3390/biotech15010013","DOIUrl":"10.3390/biotech15010013","url":null,"abstract":"<p><p>This study investigates the functional role of <i>OsHSBP1</i>, a heat shock factor-binding protein, in regulating abiotic stress tolerance in rice, with the aim of enhancing climate resilience in the elite indica cultivar Bacthom 7 (BT7). Using Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9 (CRISPR/Cas9) genome editing, we generated transgene-free homozygous knockout lines targeting <i>OsHSBP1</i> and evaluated their physiological, biochemical, and agronomic responses under heat stress. Mutant lines exhibited markedly improved tolerance to both stresses, with survival rates reaching 43-46% under heat stress, compared to near-zero in wildtype plants. Enhanced tolerance was associated with significantly increased catalase and peroxidase activities and reduced oxidative damage, including lower malondialdehyde content and decreased superoxide accumulation. Despite these stress-related advantages, the knockout lines showed minimal differences in key agronomic traits under normal growing conditions, with comparable plant height, tillering ability, grain yield, and amylose content relative to the wildtype. These results demonstrate that <i>OsHSBP1</i> functions as a negative regulator of abiotic stress tolerance in rice, and its knockout enhances resilience without compromising yield potential. The study highlights <i>OsHSBP1</i> as a promising target for precision breeding of climate-resilient rice cultivars.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"15 1","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12922113/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146259470","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}
Jonathan Puente-Rivera, Stephanie I Nuñez Olvera, Ameyatzin Ereth Robles-Chávez, Nayeli Goreti Nieto-Velázquez, María Elizbeth Alvarez-Sánchez
Prostate cancer (PCa) is a leading cause of cancer-related mortality in men and is often characterized by aggressive growth and bone metastasis. Angiogenesis plays a central role in tumor progression and dissemination. This study aimed to explore the regulatory roles of long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) in angiogenesis and metastasis during PCa progression. Publicly available RNA-seq datasets were analyzed to identify differentially expressed miRNAs between metastatic (N1) and nonmetastatic (N0) PCa. Bioinformatic tools were used to reconstruct co-regulatory networks involving miRNAs, lncRNAs, and angiogenesis-related mRNAs. RT-qPCR was performed on serum-derived liquid biopsies from N0 and N1 patients and healthy controls to validate the key regulatory axes. Transcriptomic analysis revealed that miRNAs such as hsa-miR-183-5p and hsa-miR-216a-5p were upregulated in N1 PCa and associated with pro-angiogenic signaling, whereas hsa-miR-206 and hsa-miR-184, known for their anti-angiogenic functions, were downregulated. Network analysis identified the LINC00261-miR-206-HIF1A axis as the central regulatory module. RT-qPCR validation confirmed the significant downregulation of LINC00261 and miR-206, along with HIF1A overexpression in N1 samples compared to N0 and controls (p < 0.001), supporting in silico predictions. These findings highlight the role of ncRNA-mediated regulation of PCa angiogenesis and metastasis. The LINC00261-miR-206-HIF1A axis may serve as a promising noninvasive biomarker and potential therapeutic target. The integration of computational and experimental data provides a strong rationale for the further functional validation of advanced PCa.
{"title":"Transcriptomic Insights into lncRNA-miRNA-mRNA Networks Regulating Angiogenesis and Metastasis in Prostate Cancer.","authors":"Jonathan Puente-Rivera, Stephanie I Nuñez Olvera, Ameyatzin Ereth Robles-Chávez, Nayeli Goreti Nieto-Velázquez, María Elizbeth Alvarez-Sánchez","doi":"10.3390/biotech15010012","DOIUrl":"10.3390/biotech15010012","url":null,"abstract":"<p><p>Prostate cancer (PCa) is a leading cause of cancer-related mortality in men and is often characterized by aggressive growth and bone metastasis. Angiogenesis plays a central role in tumor progression and dissemination. This study aimed to explore the regulatory roles of long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) in angiogenesis and metastasis during PCa progression. Publicly available RNA-seq datasets were analyzed to identify differentially expressed miRNAs between metastatic (N1) and nonmetastatic (N0) PCa. Bioinformatic tools were used to reconstruct co-regulatory networks involving miRNAs, lncRNAs, and angiogenesis-related mRNAs. RT-qPCR was performed on serum-derived liquid biopsies from N0 and N1 patients and healthy controls to validate the key regulatory axes. Transcriptomic analysis revealed that miRNAs such as hsa-miR-183-5p and hsa-miR-216a-5p were upregulated in N1 PCa and associated with pro-angiogenic signaling, whereas hsa-miR-206 and hsa-miR-184, known for their anti-angiogenic functions, were downregulated. Network analysis identified the LINC00261-miR-206-HIF1A axis as the central regulatory module. RT-qPCR validation confirmed the significant downregulation of LINC00261 and miR-206, along with HIF1A overexpression in N1 samples compared to N0 and controls (<i>p</i> < 0.001), supporting in silico predictions. These findings highlight the role of ncRNA-mediated regulation of PCa angiogenesis and metastasis. The LINC00261-miR-206-HIF1A axis may serve as a promising noninvasive biomarker and potential therapeutic target. The integration of computational and experimental data provides a strong rationale for the further functional validation of advanced PCa.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"15 1","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921783/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146259681","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}
Kenzhegul Bolatkhan, Ardak B Kakimova, Bolatkhan K Zayadan, Akbota Kabayeva, Sandugash K Sandybayeva, Aliyam A Dauletova, Tatsuya Tomo
The transition to low-carbon energy systems requires scalable and energy-efficient routes for producing liquid biofuels that are compatible with existing fuel infrastructures. This review focuses on bio-oil production from phototrophic microorganisms, highlighting their high biomass productivity, rapid growth, and inherent capacity for carbon dioxide fixation as key advantages over conventional biofuel feedstocks. Recent progress in thermochemical conversion technologies, particularly hydrothermal liquefaction (HTL) and fast pyrolysis, is critically assessed with respect to their suitability for wet and dry algal biomass, respectively. HTL enables direct processing of high-moisture biomass while avoiding energy-intensive drying, whereas fast pyrolysis offers high bio-oil yields from lipid-rich feedstocks. In parallel, catalytic upgrading strategies, including hydrodeoxygenation and related hydroprocessing routes, are discussed as essential steps for improving bio-oil stability, heating value, and fuel compatibility. Beyond conversion technologies, innovative biological and biotechnological strategies, such as strain optimization, stress induction, co-cultivation, and synthetic biology approaches, are examined for their role in tailoring biomass composition and enhancing bio-oil precursors. The integration of microalgal cultivation with wastewater utilization is briefly considered as a supporting strategy to reduce production costs and improve overall sustainability. Overall, this review emphasizes that the effective coupling of advanced thermochemical conversion with targeted biological optimization represents the most promising pathway for scalable bio-oil production from phototrophic microorganisms, positioning algal bio-oil as a viable contributor to future low-carbon energy systems.
{"title":"Bio-Oil from Phototrophic Microorganisms: Innovative Technologies and Strategies.","authors":"Kenzhegul Bolatkhan, Ardak B Kakimova, Bolatkhan K Zayadan, Akbota Kabayeva, Sandugash K Sandybayeva, Aliyam A Dauletova, Tatsuya Tomo","doi":"10.3390/biotech15010011","DOIUrl":"10.3390/biotech15010011","url":null,"abstract":"<p><p>The transition to low-carbon energy systems requires scalable and energy-efficient routes for producing liquid biofuels that are compatible with existing fuel infrastructures. This review focuses on bio-oil production from phototrophic microorganisms, highlighting their high biomass productivity, rapid growth, and inherent capacity for carbon dioxide fixation as key advantages over conventional biofuel feedstocks. Recent progress in thermochemical conversion technologies, particularly hydrothermal liquefaction (HTL) and fast pyrolysis, is critically assessed with respect to their suitability for wet and dry algal biomass, respectively. HTL enables direct processing of high-moisture biomass while avoiding energy-intensive drying, whereas fast pyrolysis offers high bio-oil yields from lipid-rich feedstocks. In parallel, catalytic upgrading strategies, including hydrodeoxygenation and related hydroprocessing routes, are discussed as essential steps for improving bio-oil stability, heating value, and fuel compatibility. Beyond conversion technologies, innovative biological and biotechnological strategies, such as strain optimization, stress induction, co-cultivation, and synthetic biology approaches, are examined for their role in tailoring biomass composition and enhancing bio-oil precursors. The integration of microalgal cultivation with wastewater utilization is briefly considered as a supporting strategy to reduce production costs and improve overall sustainability. Overall, this review emphasizes that the effective coupling of advanced thermochemical conversion with targeted biological optimization represents the most promising pathway for scalable bio-oil production from phototrophic microorganisms, positioning algal bio-oil as a viable contributor to future low-carbon energy systems.</p>","PeriodicalId":34490,"journal":{"name":"BioTech","volume":"15 1","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12921903/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146259422","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}