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EvoPhys-ACP: Combining evolutionary and biochemical insights for accurate anticancer peptide prediction evophyth - acp:结合进化和生化的见解,准确预测抗癌肽
IF 4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-01-01 Epub Date: 2026-02-05 DOI: 10.1016/j.crbiot.2026.100376
Sayeda Muntaha Ferdous , Shafayat Bin Shabbir Mugdha , Mahtab Uddin
Anticancer peptides (ACPs) are becoming more well-known for their ability to target only cancer cells. This ability is more effective than that of many conventional therapies, which often have numerous adverse effects. Even while they show promise, finding new ACPs through laboratory experiments is still expensive and takes a lot of time, which has slowed progress in this area. In this study, we sought to address this issue by developing a machine learning approach that predicts ACPs based on evolutionary information from Position-Specific Scoring Matrices (PSSMs) and key physicochemical properties. To refine the proposed model, we used Support Vector Machine-Recursive Feature Elimination (SVM-RFE) to select the most relevant features, and we compared several classifiers, specifically Support Vector Machines, Random Forest, and Logistic Regression, using 10-fold stratified cross-validation for a robust evaluation. Standard metrics such as accuracy, precision, recall, F1-score, MCC, and ROC-AUC were employed to evaluate predictive performance. From the achieved results, it is evident that combining evolutionary and physicochemical criteria can significantly facilitate the identification of possible ACPs. We have deployed the trained model via a local web interface to enable real-time, sequence-based prediction, facilitating the prompt application of this research. This accessible tool is positioned to catalyze further discovery and innovation in peptide-based cancer treatment development.
抗癌肽(ACPs)因其仅针对癌细胞的能力而越来越为人所知。这种能力比许多传统疗法更有效,而传统疗法往往有许多副作用。尽管它们显示出希望,但通过实验室实验发现新的acp仍然昂贵且需要大量时间,这减缓了该领域的进展。在这项研究中,我们试图通过开发一种机器学习方法来解决这个问题,该方法基于来自位置特定评分矩阵(PSSMs)和关键物理化学性质的进化信息来预测acp。为了完善所提出的模型,我们使用支持向量机递归特征消除(SVM-RFE)来选择最相关的特征,并且我们比较了几种分类器,特别是支持向量机,随机森林和逻辑回归,使用10倍分层交叉验证来进行稳健评估。准确度、精密度、召回率、f1评分、MCC和ROC-AUC等标准指标用于评估预测性能。从已经取得的结果来看,显然将进化和物理化学标准结合起来可以显著地促进可能的acp的识别。我们通过本地web界面部署了训练好的模型,实现了实时的、基于序列的预测,促进了本研究的迅速应用。这种易于使用的工具被定位为催化基于肽的癌症治疗发展的进一步发现和创新。
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引用次数: 0
CRISPR-Cas12 in immunotherapy and Beyond: Advances and challenges CRISPR-Cas12在免疫治疗及其他领域:进展与挑战
IF 4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-01-01 Epub Date: 2025-12-02 DOI: 10.1016/j.crbiot.2025.100356
Tahereh Zarei Taher , Mohammad Hassan Yousefi , Hamed Afkhami , Maria Kavianpour , Amir Hossein Kheirkhah
CRISPR-Cas12 technology has greatly advanced gene editing, offering high efficiency and accuracy. Its main applications include gene modification, disease diagnosis, and especially the development of personalized cancer therapies. CRISPR-Cas12 enhances CAR-T cell therapies, allowing immune cells to target cancer more effectively. The technology also enables precise DNA sequence changes, supporting tailored treatments based on individual tumor genetics. Importantly, CRISPR-Cas12 reduces off-target effects, improving the safety and reliability of gene therapies in clinical settings. Additionally, strategies to overcome the immunosuppressive tumor microenvironment, such as modulating regulatory T cells or engineering resistance to their inhibitory signals, are emphasized. However, challenges remain, including concerns about long-term safety, delivery methods, and ethical issues related to genetic modification. Despite these hurdles, CRISPR-Cas12 offers significant potential as a safe and transformative tool for innovative immunotherapies, promising more effective cancer treatments and better patient outcomes.
CRISPR-Cas12技术极大地推进了基因编辑,具有高效率和准确性。它的主要应用包括基因修饰、疾病诊断,特别是个性化癌症治疗的发展。CRISPR-Cas12增强了CAR-T细胞疗法,使免疫细胞更有效地靶向癌症。该技术还可以实现精确的DNA序列改变,支持基于个体肿瘤遗传学的定制治疗。重要的是,CRISPR-Cas12减少了脱靶效应,提高了临床环境中基因治疗的安全性和可靠性。此外,还强调了克服免疫抑制肿瘤微环境的策略,如调节调节性T细胞或对其抑制信号的工程抗性。然而,挑战依然存在,包括对长期安全性、递送方法和与转基因相关的伦理问题的担忧。尽管存在这些障碍,CRISPR-Cas12作为创新免疫疗法的安全和变革性工具提供了巨大的潜力,有望提供更有效的癌症治疗和更好的患者预后。
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引用次数: 0
Acoustic signatures of microalgal growth: ultrasonic velocity and attenuation as quantitative biomass indicators in Spirulina maxima 微藻生长的声学特征:超声速度和衰减作为最大螺旋藻生物量的定量指标
IF 4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-01-01 Epub Date: 2026-03-07 DOI: 10.1016/j.crbiot.2026.100380
Darmawan Hidayat , Hilman Sya’ban Sulthoni , Taufik Taufik , Septian Ari Kurniawan , Muhammad Rasyid Ramdhani , Asri Peni Wulandari
Accurate, real-time measurement of microalgal biomass is vital for optimizing photobioreactor performance and advancing bioprocess monitoring technologies. This study systematically investigated the acoustic signatures of Spirulina maxima (S. maxima) by measuring sound velocity and attenuation at 1 MHz over a 14-day cultivation period and across controlled biomass dilutions (0–100% v/v). Ultrasonic time-of-flight and amplitude measurements were used to determine acoustic parameters, which were subsequently correlated with optical density (OD680) to develop predictive models for biomass estimation. Sound velocity increased from approximately 1507 to 1710 m/s during cultivation, exhibiting nonlinear, concentration-dependent behavior (R2 = 0.96), while attenuation rose from 0.0035 to 0.0074 dB/cm and exhibited near-linear dependence at early stages, with pronounced nonlinearity at higher biomass levels (R2 = 0.93). Concentration-resolved measurements confirmed consistent trends: sound velocity increased steadily across all biomass concentrations, whereas attenuation increased sharply beyond roughly 60% v/v due to greater scattering and viscous–thermal losses. Temporal OD680 profiles revealed distinct lag, quasi-stationary, and secondary growth phases, each reflected in characteristic shifts in acoustic response. Strong correlations between ultrasonic parameters and OD680 (velocity: r = 0.93; attenuation: r = 0.97) demonstrate the capability of acoustic sensing to detect both mechanical and structural changes in microalgal cultures. Overall, these findings establish low-power ultrasound as a robust, sensitive, and non-invasive technique for real-time biomass monitoring, with significant potential for integration into automated photobioreactors and environmental monitoring systems.
准确、实时测量微藻生物量对于优化光生物反应器性能和推进生物过程监测技术至关重要。本研究系统地研究了最大螺旋藻(S. maxima)的声学特征,在14天的培养周期内,通过控制生物量稀释(0-100% v/v)测量1 MHz声速和衰减。超声飞行时间和振幅测量用于确定声学参数,随后将声学参数与光密度(OD680)相关联,以建立生物量估算的预测模型。在培养过程中,声速从1507 ~ 1710 m/s增加,表现出非线性的浓度依赖关系(R2 = 0.96);衰减从0.0035 ~ 0.0074 dB/cm增加,早期表现出近似线性的依赖关系(R2 = 0.93)。浓度分辨测量证实了一致的趋势:声速在所有生物量浓度下都稳定增加,而衰减在大约60% v/v以上急剧增加,这是由于更大的散射和粘热损失。OD680的时间分布显示出明显的滞后、准平稳和二次生长阶段,每个阶段都反映在声响应的特征变化中。超声参数与OD680之间的强相关性(速度:r = 0.93;衰减:r = 0.97)表明声学传感能够检测微藻培养物的力学和结构变化。总的来说,这些发现确立了低功率超声作为一种强大、灵敏、无创的实时生物质监测技术,具有集成到自动化光生物反应器和环境监测系统中的巨大潜力。
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引用次数: 0
Plant vs mammal extracellular vesicles: new tools in therapeutic drug delivery 植物与哺乳动物的细胞外囊泡:治疗药物输送的新工具
IF 4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2026-01-01 Epub Date: 2025-11-21 DOI: 10.1016/j.crbiot.2025.100352
Elisa Maricchiolo, Pasquale Creanza, Michela Osnato, Mattia Tiboni, Luca Casettari, Annalisa Aluigi, Andrea Pompa
Extracellular vesicles (EVs) are highly promising biological carriers, due to their potential for drug delivery and their involvement in multiple cellular processes. These vesicles, released by cells into the extracellular space, can be classified into different types based on their origin, size, and molecular content. While animal-derived EVs have been extensively studied and characterised, recent research has highlighted the growing interest in plant-derived EVs, which present unique properties that could offer novel advantages in therapeutic applications.
This review aims to provide an overview of similarities and differences between animal and plant-derived EVs, focusing on their structural and functional characteristics. Additionally, we report various methods used for the isolation and characterisation of these vesicles, to provide a comprehensive summary, highlighting the main challenges and opportunities of each experimental option.
Moreover, the present work explores the most used techniques for loading bioactive molecules into EVs, transforming them from natural biological entities into innovative drug delivery systems. The ability to encapsulate drugs, small RNA molecules, or proteins within EVs opens up new frontiers for the targeted treatment of diseases, with particular emphasis on overcoming barriers related to drug bioavailability and specificity.
Finally, this review stresses the importance of ongoing research in the extracellular vesicles field, from both animal and plant origins, given their adaptable properties to multiple pharmaceutical purposes. Plant-derived EVsoffer a sustainable and economic alternative, together with the better characterised mammalian vesicles, showing the potential to strongly improve the drug delivery strategy in the near future.
细胞外囊泡(EVs)是非常有前途的生物载体,因为它们具有潜在的药物递送和参与多种细胞过程。这些囊泡由细胞释放到细胞外空间,根据它们的来源、大小和分子含量可分为不同类型。虽然动物源性电动汽车已经被广泛研究和表征,但最近的研究强调了对植物源性电动汽车日益增长的兴趣,因为植物源性电动汽车具有独特的特性,可以在治疗应用中提供新的优势。本文综述了动物源性电动汽车和植物源性电动汽车的异同,重点介绍了它们的结构和功能特征。此外,我们报告了用于分离和表征这些囊泡的各种方法,以提供一个全面的总结,突出每个实验选项的主要挑战和机遇。此外,本研究还探索了将生物活性分子装载到电动汽车中,将其从天然生物实体转化为创新的药物输送系统的最常用技术。在ev内封装药物、小RNA分子或蛋白质的能力为疾病的靶向治疗开辟了新的领域,特别强调克服与药物生物利用度和特异性相关的障碍。最后,本综述强调了细胞外囊泡领域正在进行的研究的重要性,从动物和植物的起源,考虑到它们对多种药物用途的适应性。植物源性ev软膏是一种可持续和经济的替代品,与更好表征的哺乳动物囊泡一起,显示出在不久的将来强有力地改善药物递送策略的潜力。
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引用次数: 0
The landscape of LncRNAs in diabetic kidney disease: a meta-analysis of transcriptomics data lncrna在糖尿病肾病中的作用:转录组学数据的荟萃分析
IF 4 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-07-30 DOI: 10.1016/j.crbiot.2025.100322
Raziyeh Rezaei , Basireh Bahrami , Yousof Gheisari
Non-coding regions of the genome are known to influence complex disorders, yet the role of long non-coding RNAs (lncRNAs) in Diabetic Kidney Disease (DKD) remains underexplored. This study conducts a meta-analysis of RNA-sequencing data from murine kidney samples of type 1 (T1DM) and type 2 diabetes mellitus (T2DM) to identify lncRNAs associated with DKD. DKD-associated datasets were harvested, and after data pre-processing and quality assessment, 6 T1DM and 4 T2DM datasets were included. Data integration, batch correction, and normalization were performed, followed by the identification of differentially expressed lncRNAs (meta-DELs) and mRNAs (meta-DEMs). A DKD mouse model was developed to validate the expression of selected meta-DELs using qRT-PCR. The meta-analysis identified 188 meta-DELs in T1DM and 68 in T2DM. Notably, a small set of lncRNAs have dense mRNA interactions, including Dancer, Gm7628, C4a, and Gm17300 in T1DM and Malat1, C4a, Gm17300, and Eif4a2 in T2DM. Expression analysis confirmed the up-regulation of seven selected meta-DELs in the DKD model, with Trp53cor1, Gm15462, and Gm42664 reaching statistical significance. This systematic analysis of high-quality expression profiles identified meta-DELs consistently associated with DKD, distinguishing actual lncRNA changes from those influenced by experimental conditions or gene expression noise.
众所周知,基因组的非编码区会影响复杂的疾病,但长链非编码rna (lncRNAs)在糖尿病肾病(DKD)中的作用仍未得到充分探索。本研究对1型(T1DM)和2型糖尿病(T2DM)小鼠肾脏样本的rna测序数据进行了荟萃分析,以确定与DKD相关的lncrna。收集与dkd相关的数据集,经过数据预处理和质量评估,包括6个T1DM和4个T2DM数据集。进行数据整合、批量校正和归一化,随后鉴定差异表达的lncrna (meta-DELs)和mrna (meta- dem)。建立DKD小鼠模型,利用qRT-PCR验证所选meta-DELs的表达。荟萃分析发现,T1DM患者有188例,T2DM患者有68例。值得注意的是,一小部分lncrna具有密集的mRNA相互作用,包括T1DM中的Dancer、Gm7628、C4a和Gm17300,以及T2DM中的Malat1、C4a、Gm17300和Eif4a2。表达分析证实了DKD模型中7个选定的meta-DELs的上调,其中Trp53cor1、Gm15462和Gm42664达到了统计学意义。这项高质量表达谱的系统分析发现了与DKD一致相关的meta- del,将实际的lncRNA变化与受实验条件或基因表达噪声影响的lncRNA变化区分开来。
{"title":"The landscape of LncRNAs in diabetic kidney disease: a meta-analysis of transcriptomics data","authors":"Raziyeh Rezaei ,&nbsp;Basireh Bahrami ,&nbsp;Yousof Gheisari","doi":"10.1016/j.crbiot.2025.100322","DOIUrl":"10.1016/j.crbiot.2025.100322","url":null,"abstract":"<div><div>Non-coding regions of the genome are known to influence complex disorders, yet the role of long non-coding RNAs (lncRNAs) in Diabetic Kidney Disease (DKD) remains underexplored. This study conducts a meta-analysis of RNA-sequencing data from murine kidney samples of type 1 (T1DM) and type 2 diabetes mellitus (T2DM) to identify lncRNAs associated with DKD. DKD-associated datasets were harvested, and after data pre-processing and quality assessment, 6 T1DM and 4 T2DM datasets were included. Data integration, batch correction, and normalization were performed, followed by the identification of differentially expressed lncRNAs (meta-DELs) and mRNAs (meta-DEMs). A DKD mouse model was developed to validate the expression of selected meta-DELs using qRT-PCR. The meta-analysis identified 188 meta-DELs in T1DM and 68 in T2DM. Notably, a small set of lncRNAs have dense mRNA interactions, including <em>Dancer</em>, <em>Gm7628</em>, <em>C4a</em>, and <em>Gm17300</em> in T1DM and <em>Malat1</em>, <em>C4a</em>, <em>Gm17300</em>, and <em>Eif4a2</em> in T2DM. Expression analysis confirmed the up-regulation of seven selected meta-DELs in the DKD model, with Trp53cor1, Gm15462, and Gm42664 reaching statistical significance. This systematic analysis of high-quality expression profiles identified meta-DELs consistently associated with DKD, distinguishing actual lncRNA changes from those influenced by experimental conditions or gene expression noise.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"10 ","pages":"Article 100322"},"PeriodicalIF":4.0,"publicationDate":"2025-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145003841","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Validation of microbial source tracking markers through PCR-based molecular analysis and microbial genome database 通过基于pcr的分子分析和微生物基因组数据库验证微生物源跟踪标记
IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-01-01 Epub Date: 2025-06-02 DOI: 10.1016/j.crbiot.2025.100304
Rifat Zubair Ahmed , Ashraful Islam , Tiantian Tian , Zhe Tian , Yu Zhang , Min Yang
Microbial Source Tracking (MST) uses molecular markers targeting host-associated gut microorganisms to identify fecal pollution. However, MST faces significant challenges in fecal source identification, particularly due to the markers’ poor specificity and shared genomic areas among microorganisms from different host sources. This study addresses these challenges by using host-specific Escherichia coli genetic markers, originally developed through a novel, library-independent approach, to detect sources of fecal pollution. A total of 563 E.coli isolates from chicken, cow, and pig feces were isolated and assessed by nine reported host-associated E. coli genetic markers (Chicken: CH7, CH9, CH12, CH13; Cow: CO2, CO3; Pig: P1, P3, P4) through PCR. Marker possession patterns, sensitivity, specificity, and accuracy were calculated. The NCBI Microbial Genome database was searched for sequences homologous to genome regions of studied genetic markers and evaluated by finding the percentage of host sources and sequence location in the genome. Homology evaluation with binary PCR results was used to predict the best-performing marker. PCR results exhibited that the most effective markers were chicken CH7 (67% sensitivity, 77.9% specificity, 74.4% accuracy) and CH9 (55% sensitivity, 99.4% specificity, 84.7% accuracy). However, a homology search in the database narrowed the selection of the top-performing marker to CH7, which showed homology with E.coli from chicken hosts, while other markers exhibited higher homology with E.coli from Humans. Furthermore, sequences from the database homologous to the CH9 and CO2 markers were found on a plasmid, while those for CH12, CO3, P1, and P4 were on the chromosome, and CH7, CH13, and P3 were on both. This study highlights the critical need for integrated approaches to assess molecular markers in MST assays, emphasizing their significance in advancing research within the field.
微生物源追踪技术(MST)是一种利用分子标记靶向宿主相关肠道微生物来识别粪便污染的技术。然而,MST在粪便来源鉴定方面面临着重大挑战,特别是由于标记的特异性较差,并且来自不同宿主来源的微生物具有相同的基因组区域。本研究通过使用宿主特异性大肠杆菌遗传标记来解决这些挑战,该标记最初是通过一种新颖的、独立于文库的方法开发的,用于检测粪便污染源。从鸡、牛和猪粪便中分离出563株大肠杆菌,并利用9个已报道的宿主相关大肠杆菌遗传标记(鸡:CH7、CH9、CH12、CH13;牛:CO2, CO3;猪:P1, P3, P4)。计算标记物占有模式、敏感性、特异性和准确性。在NCBI微生物基因组数据库中搜索与所研究遗传标记的基因组区域同源的序列,并通过寻找宿主来源的百分比和序列在基因组中的位置进行评估。用二元PCR结果进行同源性评价,预测最佳标记。PCR结果显示,鸡CH7(敏感性67%,特异性77.9%,准确性74.4%)和CH9(敏感性55%,特异性99.4%,准确性84.7%)是最有效的标记。然而,在数据库中进行同源性搜索,将选择的最佳标记缩小到CH7,该标记与鸡宿主大肠杆菌具有同源性,而其他标记与人类大肠杆菌具有较高的同源性。此外,在一个质粒上发现了数据库中与CH9和CO2标记同源的序列,而CH12、CO3、P1和P4标记在染色体上,CH7、CH13和P3标记在两条染色体上。这项研究强调了在MST检测中评估分子标记的综合方法的迫切需要,强调了它们在推进该领域研究中的重要性。
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引用次数: 0
Gut health improvement by locally isolated probiotics and histomorphometric analysis in Wistar rats 局部分离益生菌改善Wistar大鼠肠道健康及组织形态计量学分析
IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-01-01 Epub Date: 2025-01-08 DOI: 10.1016/j.crbiot.2024.100271
Zuhra Bibi , Dilara Abbas Bukhari , Muhammad Qadeer Sarwar , Arifullah , Samina Younas , Tayyab Manzoor , Abdul Rehman
In the present investigation, lab-isolated probiotics Weisella confusa MZ735961.1, Lactiplantibacillus plantarum MZ707748.1, L. plantarum MZ710117.1, and L. plantarum MZ735961 were used separately and in combinations to evaluate their effect on gut morphology of Wistar rats. Synergistic groups were formed by 1:1 and labeled as G1 (L. plantarum MZ707748.1 and L. plantarum MZ729681.1), G2 (W. confusa MZ735961.1 and L. plantarum MZ727611.1), G3 (L. plantarum MZ729681.1, W. confusa MZ735961.1, and Lactobacillus acidophilus La-14), G4 (all above mentioned probiotics). Rats were gavage-fed with probiotics according to their colony-forming unit (CFU). The experiment was carried out for 35 days. The bacteria were re-isolated from the gut and identified by biochemical tests which confirmed the administration and re-isolation of different Lactobacillus strains from the gut. Molecular characterization was done through 16S rRNA by using universal primers. After sequencing eight Lactobacillus strains were identified. Histopathology of rats’ intestines was done, and different parameters were examined. Villus height, crypt height, crypt width, mucosa, and sub-mucosa of jejunum were significantly (p = 0.00) increased in the G3 synergetic probiotic group compared to 0-day and negative control. However, the villus width showed non-significant (p > 0.05) variations in both genders. Mucosa tunic, muscle tunic, total wall, and crypt depth were significantly increased (p = 0.00) in the G4 group of medial colon. The study concluded that gut morphology improves as probiotics adhere better to the intestinal epithelium, excluding pathogens, reducing inflammation, enhancing nutrient absorption, and stimulating mucosal growth. This results in improved villus structure and gut wall integrity.
本研究采用实验室分离的几种益生菌,分别使用杂交魏氏菌MZ735961.1、植物乳杆菌MZ707748.1、植物乳杆菌MZ710117.1和植物乳杆菌MZ735961,观察其对Wistar大鼠肠道形态的影响。按1:1组成协同组,分别标记为G1 (L. plantarum MZ707748.1和L. plantarum MZ729681.1)、G2 (W. confusa MZ735961.1和L. plantarum MZ727611.1)、G3 (L. plantarum MZ729681.1、W. confusa MZ735961.1和嗜酸乳杆菌La-14)、G4(上述益生菌)。大鼠按菌落形成单位(CFU)灌喂益生菌。试验期35 d。从肠道中重新分离细菌,并通过生化试验鉴定,证实了肠道中不同乳杆菌菌株的给药和重新分离。使用通用引物通过16S rRNA进行分子鉴定。测序后鉴定出8株乳杆菌。对大鼠肠道进行组织病理学检查,并对各参数进行检测。与0日龄和阴性对照组相比,G3协同益生菌组空肠绒毛高度、隐窝高度、隐窝宽度、黏膜和亚黏膜显著(p = 0.00)升高。但绒毛宽度无显著差异(p >;0.05)的差异。G4组内结肠粘膜、肌被膜、总肠壁、隐窝深度均显著增加(p = 0.00)。该研究得出结论,肠道形态改善,因为益生菌更好地粘附在肠上皮上,排除病原体,减少炎症,促进营养吸收,刺激粘膜生长。这改善了绒毛结构和肠壁的完整性。
{"title":"Gut health improvement by locally isolated probiotics and histomorphometric analysis in Wistar rats","authors":"Zuhra Bibi ,&nbsp;Dilara Abbas Bukhari ,&nbsp;Muhammad Qadeer Sarwar ,&nbsp;Arifullah ,&nbsp;Samina Younas ,&nbsp;Tayyab Manzoor ,&nbsp;Abdul Rehman","doi":"10.1016/j.crbiot.2024.100271","DOIUrl":"10.1016/j.crbiot.2024.100271","url":null,"abstract":"<div><div>In the present investigation, lab-isolated probiotics <em>Weisella confusa</em> MZ735961.1, <em>Lactiplantibacillus plantarum</em> MZ707748.1<em>, L. plantarum</em> MZ710117.1<em>,</em> and <em>L. plantarum</em> MZ735961 were used separately and in combinations to evaluate their effect on gut morphology of Wistar rats. Synergistic groups were formed by 1:1 and labeled as G1 (<em>L. plantarum</em> MZ707748.1 and <em>L. plantarum</em> MZ729681.1), G2 (<em>W. confusa</em> MZ735961.1 and <em>L. plantarum</em> MZ727611.1), G3 (<em>L. plantarum</em> MZ729681.1, <em>W. confusa</em> MZ735961.1, and <em>Lactobacillus acidophilus</em> La-14), G4 (all above mentioned probiotics). Rats were gavage-fed with probiotics according to their colony-forming unit (CFU). The experiment was carried out for 35 days. The bacteria were re-isolated from the gut and identified by biochemical tests which confirmed the administration and re-isolation of different <em>Lactobacillus</em> strains from the gut. Molecular characterization was done through 16S rRNA by using universal primers. After sequencing eight <em>Lactobacillus</em> strains were identified. Histopathology of rats’ intestines was done, and different parameters were examined. Villus height, crypt height, crypt width, mucosa, and sub-mucosa of jejunum were significantly (p = 0.00) increased in the G3 synergetic probiotic group compared to 0-day and negative control. However, the villus width showed non-significant (p &gt; 0.05) variations in both genders. Mucosa tunic, muscle tunic, total wall, and crypt depth were significantly increased (p = 0.00) in the G4 group of medial colon. The study concluded that gut morphology improves as probiotics adhere better to the intestinal epithelium, excluding pathogens, reducing inflammation, enhancing nutrient absorption, and stimulating mucosal growth. This results in improved villus structure and gut wall integrity.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"9 ","pages":"Article 100271"},"PeriodicalIF":3.6,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143136459","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}
引用次数: 0
Cost-effective production process of scFv antibody fragments against Shiga toxin 2 via recombinant E. coli 利用重组大肠杆菌生产抗志贺毒素2单链病毒抗体片段的成本效益研究
IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-01-01 Epub Date: 2025-06-30 DOI: 10.1016/j.crbiot.2025.100310
Marcela Guimarães , Daniela Luz , Elisabeth de Fátima Pires Augusto , Lucia Vieira , Maricilia Silva Costa , Roxane Maria Fontes Piazza , José Geraldo da Cruz Pradella
Shiga toxin (Stx)-producing Escherichia coli (STEC) and its subgroup enterohemorrhagic E. coli are significant pathogens responsible for diarrhea, which can progress to hemorrhagic colitis and hemolytic uremic syndrome (HUS), the leading cause of acute renal failure in children. Early diagnosis is crucial for effective clinical management, as antibiotic treatment is not recommended for STEC infections. The present study aimed to establish a cost-effective biotechnological platform for cultivating recombinant E. coli to produce scFv antibody fragments against Stx2 for diagnostic applications. The method was first evaluated through shake flask experiments and subsequently scaled up to bench-scale bioreactors operated in both batch and fed-batch modes using defined culture media. Optimal production conditions were achieved by inducing recombinant E. coli pLys at 18 °C for 18 h with 0.1 mM IPTG, resulting in a yield of 3.0 to 4.0 mg scFv/g cell biomass. A fed-batch, high-cell-density procedure with E. coli pLysS achieved a maximum production up to 150 mg scFv/L. A preliminary economic assessment demonstrated the production potential at a value of around $250/g scFv. Economic analysis also highlights that the relative cost of capital investment becomes important as production processes intensify. Therefore, technical parameters such as productivity (scFv mass/bioreactor volume * time) and scFv concentration (mass scFv mass/bioreactor volume) should be prioritized to maximize their values. Similarly, optimization of the recombinant E. coli microbial platform should be pursued to increase the Yp/x level.
产志贺毒素(Stx)的大肠杆菌(STEC)及其亚群肠出血性大肠杆菌是导致腹泻的重要病原体,可发展为出血性结肠炎和溶血性尿毒症综合征(HUS),是儿童急性肾衰竭的主要原因。早期诊断对于有效的临床管理至关重要,因为不建议对产志毒素大肠杆菌感染进行抗生素治疗。本研究旨在建立一个具有成本效益的培养重组大肠杆菌的生物技术平台,以生产用于诊断的Stx2的scFv抗体片段。该方法首先通过摇瓶实验进行评估,随后扩展到使用确定的培养基在批处理和补料批处理模式下运行的实验规模生物反应器。最佳的生产条件是在18°C下,用0.1 mM IPTG诱导重组大肠杆菌pLys 18 h,产量为3.0 ~ 4.0 mg scFv/g细胞生物量。使用大肠杆菌pLysS进行分批喂料、高密度处理,最大产量可达150 mg scFv/L。初步经济评估表明,其生产潜力约为250美元/克scFv。经济分析还强调,随着生产过程的加剧,资本投资的相对成本变得越来越重要。因此,应优先考虑生产效率(scFv质量/生物反应器体积*时间)和scFv浓度(质量scFv质量/生物反应器体积)等技术参数,使其值最大化。同样,重组大肠杆菌的微生物平台也需要优化,以提高Yp/x水平。
{"title":"Cost-effective production process of scFv antibody fragments against Shiga toxin 2 via recombinant E. coli","authors":"Marcela Guimarães ,&nbsp;Daniela Luz ,&nbsp;Elisabeth de Fátima Pires Augusto ,&nbsp;Lucia Vieira ,&nbsp;Maricilia Silva Costa ,&nbsp;Roxane Maria Fontes Piazza ,&nbsp;José Geraldo da Cruz Pradella","doi":"10.1016/j.crbiot.2025.100310","DOIUrl":"10.1016/j.crbiot.2025.100310","url":null,"abstract":"<div><div>Shiga toxin (Stx)-producing <em>Escherichia coli</em> (STEC) and its subgroup enterohemorrhagic <em>E. coli</em> are significant pathogens responsible for diarrhea, which can progress to hemorrhagic colitis and hemolytic uremic syndrome (HUS), the leading cause of acute renal failure in children. Early diagnosis is crucial for effective clinical management, as antibiotic treatment is not recommended for STEC infections. The present study aimed to establish a cost-effective biotechnological platform for cultivating recombinant <em>E. coli</em> to produce scFv antibody fragments against Stx2 for diagnostic applications. The method was first evaluated through shake flask experiments and subsequently scaled up to bench-scale bioreactors operated in both batch and fed-batch modes using defined culture media. Optimal production conditions were achieved by inducing recombinant <em>E. coli</em> pLys at 18 °C for 18 h with 0.1 mM IPTG, resulting in a yield of 3.0 to 4.0 mg scFv/g cell biomass. A fed-batch, high-cell-density procedure with <em>E. coli</em> pLysS achieved a maximum production up to 150 mg scFv/L. A preliminary economic assessment demonstrated the production potential at a value of around $250/g scFv. Economic analysis also highlights that the relative cost of capital investment becomes important as production processes intensify. Therefore, technical parameters such as productivity (scFv mass/bioreactor volume * time) and scFv concentration (mass scFv mass/bioreactor volume) should be prioritized to maximize their values. Similarly, optimization of the recombinant <em>E. coli</em> microbial platform should be pursued to increase the Yp/x level.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"10 ","pages":"Article 100310"},"PeriodicalIF":3.6,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144549698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Bioremediation of heavy metals from electronic waste dumping sites with bacteria 利用细菌对电子废弃物倾倒场重金属进行生物修复
IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-01-01 Epub Date: 2025-06-21 DOI: 10.1016/j.crbiot.2025.100309
Husnain Ahmad Khan , Shahid Sher , Dilara Abbas Bukhari , Abdul Rehman
Samples were collected from two e-waste dumping sites (Mehmood Booti (31°36′28″N, 74°23′36″E) and Lakhodair (31°37′36.6″ N, 74°25′07.6″ E)) in Lahore, Pakistan. A portable multiparameter was used to determine physicochemical parameters such as temperature, pH, electrical conductivity, turbidity, total suspended particles, and total dissolved solids. Minimal salt broth was used for the determination of the minimal inhibitory concentration of the bacterium against all heavy metals. Bacterial morphology was observed under a scanning electron microscope with and without metal stress. The temperature range for all these samples was 28.7 to 35.7 °C, while the pH range was 6.7 to 7.89. The other parameters range, such as electrical conductivity µS/cm (698–8742), turbidity (14.2–103), total suspended particles (31–698), and total dissolved solids (564–23456). The lead concentration in the Mehmood Booti soil sample was 1800 mg/kg, while in the Lakhodair soil, it was 1567 mg/kg. Microbacterium sp. strain 1S1 was utilized for bioremediation assay at the lab and pilot scale. The resistance capacity of this bacterium against different metals was in the following order: As > Pb > Cd > Cu > Cr > Ni. The bioremediation potential of the bacterium against arsenic was 81.33 % and 96 % after 2 and 4 days. The least activity was observed against nickel, which was 17 and 28.33 % after 2 and 4 days. The metal removal capacity per CFU was the maximum for lead and arsenic compared to other metals, which were 1.99E-7 and 1.45E-07. The heat-inactivated bacterial cells removed arsenic in higher concentrations and lead in lower concentrations. The electron microscopy showed no significant alteration in bacterial morphology in control and metal-treated bacterial cells. The nanopore long-read sequencing analysis revealed that cadmium, nickel, copper, and arsenic resistance genes were found on the bacterial genome. No genes were found for lead and chromium but 849 hypothetical coding sequences having unknown functions were present on the bacterial genome. So, the Microbacterium sp. strain 1S1 is a potential candidate for the removal of heavy metals from e-waste dumping sites.
样本采集自巴基斯坦拉合尔的两个电子垃圾倾倒场(Mehmood Booti(31°36′28″N, 74°23′36″E)和Lakhodair(31°37′36.6″N, 74°25′07.6″E))。便携式多参数仪用于测定理化参数,如温度、pH、电导率、浊度、总悬浮颗粒和总溶解固体。用微量盐肉汤测定细菌对所有重金属的最低抑菌浓度。在扫描电子显微镜下观察有无金属应力的细菌形态。所有样品的温度范围为28.7 ~ 35.7℃,pH范围为6.7 ~ 7.89。其他参数范围,如电导率µS/cm(698-8742),浊度(14.2-103),总悬浮颗粒(31-698)和总溶解固体(564-23456)。Mehmood Booti土壤样品的铅浓度为1800 mg/kg, Lakhodair土壤样品的铅浓度为1567 mg/kg。在实验室和中试规模上采用微杆菌1S1菌株进行生物修复试验。该细菌对不同金属的抗性强弱顺序为:As >;Pb祝辞Cd比;铜比;Cr祝辞倪。2 d和4 d后,细菌对砷的生物修复率分别为81.33%和96%。2 d和4 d对镍的活性最低,分别为17%和28.33%。每CFU对铅和砷的金属去除率最高,分别为1.99E-7和1.45E-07。热灭活的细菌细胞去除高浓度的砷和低浓度的铅。电镜观察显示,对照组和金属处理过的细菌细胞形态无明显变化。纳米孔长读测序分析显示,在细菌基因组上发现了抗镉、抗镍、抗铜和抗砷基因。没有发现铅和铬的基因,但在细菌基因组中存在849个功能未知的假设编码序列。因此,微细菌sp.菌株1S1是去除电子垃圾倾倒场重金属的潜在候选者。
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引用次数: 0
Corrigendum to “Postbiotics, a natural feed additive for growth performance, gut microbiota and quality of poultry products” [Curr. Res. Biotechnol. 8 (2024) 100247] 更正:"益生菌,一种促进家禽生长性能、肠道微生物群和产品质量的天然饲料添加剂"[Curr. Res. Biotechnol.
IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2025-01-01 Epub Date: 2025-04-13 DOI: 10.1016/j.crbiot.2025.100284
Jakub Urban , Karwan Yaseen Kareem , Atanas G. Atanasov , Arkadiusz Matuszewski , Damian Bień , Patrycja Ciborowska , Anna Rygało-Galewska , Monika Michalczuk
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引用次数: 0
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Current Research in Biotechnology
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