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Comparative evaluation of E. coli expression systems for soluble hScFv-IL-17A with an unpaired CDR-L3 cysteine 可溶性hScFv-IL-17A与未配对CDR-L3半胱氨酸的大肠杆菌表达系统的比较评价
IF 2.8 Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2025-12-01 Epub Date: 2025-11-07 DOI: 10.1016/j.jgeb.2025.100613
Weeraya Thongkum , Kanokporn Sornsuwan , Thanathat Pamonsupornwichit , Anuwat Weechan , Natsima Viriyaadhammaa , Kanchanok Kodchakorn , Chatchai Tawapiwatana
This study systematically evaluated Escherichia coli (E. coli) expression systems for the production of a humanized single-chain variable fragment targeting interleukin-17A (hScFv-IL-17A). A wild-type hScFv (hScFv-IL-17A-WT), derived from Secukinumab, was compared with a C97S mutant (hScFv-IL-17A-C97S) in which the unpaired cysteine residue in the light-chain complementarity-determining region 3 (CDR3) was substituted with serine. The primary objectives were to identify an optimal expression system for soluble protein production and to assess the functional consequences of the C97S mutation. Protein solubility, yield, and antigen-binding properties were evaluated across expression on three E. coli strains: Origami B (DE3), SHuffle, and BL21 (DE3) incorporating the DisCoTune system. Analyses were performed using SDS-PAGE, Western blotting, enzyme-linked immunosorbent assay (ELISA), and biolayer interferometry (BLI). Among the systems tested, BL21 (DE3) with DisCoTune yielded the highest levels of soluble expression for both variants. Although the C97S mutation enhanced soluble protein yield in this system, it significantly impaired functional activity. The hScFv-IL-17A-WT exhibited a strong binding affinity (KD = 3.64 × 10−8 M), whereas the hScFv-IL-17A-C97S demonstrated an approximately 10-fold reduction in affinity. Competitive ELISA further confirmed that both variants recognized the same epitope as a neutralizing monoclonal antibody. Structural modeling predicted that the C97S substitution imposed a moderate stability penalty without inducing major conformational alterations, consistent with the observed reduction in binding affinity. Collectively, these findings indicate that while the C97S mutation improves soluble expression, it compromises biological activity. The hScFv-IL-17A-WT produced in BL21 (DE3) using the DisCoTune system represents the most promising candidate for subsequent therapeutic development.
本研究系统地评估了大肠杆菌(E. coli)表达系统产生针对白细胞介素- 17a (hScFv-IL-17A)的人源化单链可变片段。来源于Secukinumab的野生型hScFv (hScFv- il - 17a - wt)与C97S突变体(hScFv- il - 17a -C97S)进行了比较,其中轻链互补决定区3 (CDR3)中的未配对半胱氨酸残基被丝氨酸取代。主要目的是确定可溶性蛋白生产的最佳表达系统,并评估C97S突变的功能后果。蛋白溶解度、产量和抗原结合特性在三种大肠杆菌菌株Origami B (DE3)、SHuffle和BL21 (DE3)上进行了评估。采用SDS-PAGE、Western blotting、酶联免疫吸附法(ELISA)和生物层干涉法(BLI)进行分析。在测试的系统中,BL21 (DE3)与distune在两种变体中均产生最高水平的可溶性表达。虽然C97S突变提高了该系统的可溶性蛋白产量,但显著损害了该系统的功能活性。hScFv-IL-17A-WT表现出很强的结合亲和力(KD = 3.64 × 10−8 M),而hScFv-IL-17A-C97S表现出大约10倍的亲和力降低。竞争性ELISA进一步证实,这两种变体识别相同的表位作为中和单克隆抗体。结构模型预测,C97S取代造成了适度的稳定性损失,而不会引起主要的构象改变,这与观察到的结合亲和力降低一致。总的来说,这些发现表明,虽然C97S突变提高了可溶性表达,但它损害了生物活性。使用distune系统在BL21 (DE3)中产生的hScFv-IL-17A-WT代表了后续治疗开发中最有希望的候选物。
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引用次数: 0
Eco-friendly synthesis of silver nanoparticles from the leaf blades extract of seagrass Thalassia hemprichii: Multifunctional agents for biomedical and environmental applications 从海草叶片提取物中生态友好地合成纳米银:生物医学和环境应用的多功能剂
IF 2.8 Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2025-12-01 Epub Date: 2025-10-22 DOI: 10.1016/j.jgeb.2025.100600
Shibin Eranhottu , Tijo Cherian , R. Mohanraju , N. Sharmila Devi , Jayalakshmi Sanal , Lincy Sara Varghese , P. Ambili , Mini Thomas , Fahmeeda Parveen P.S. , Willie J.G.M. Peijnenburg
The current study reports for the first time the synthesis of environmentally friendly silver nanoparticles (AgNPs) using leaf blades extract (THE) of seagrass Thalassia hemprichii. The AgNPs were described using several spectroscopic methods. The UV–visible spectral study of THE-AgNPs reported absorbance peak at 420 nm. The intrinsic stretching was seen in the FT-IR (Fourier transform infrared) spectrum inferring the presence and role of diverse functional species in the biological synthesis of AgNPs. The AgNPs reported uniform morphology, spherical shape and high stability validated by electron microscopic and zeta potential studies. The biosynthesized silver nanoparticles were employed in experiments including antibacterial, anti-diabetic (α-amylase (77%), α-glucosidase (78%) at 30 μg/ml, antioxidant (85–90%), anti-inflammatory (45–60%) properties and dye degrading properties. The antibacterial efficacy of silver nanoparticles was assessed against pathogenic species of Staphylococcus aureus and Escherichia coli reporting inhibitory zones of 13 ± 0.3 and 21 ± 0.2 mm, respectively comparable to the standard Gentamycin (22 ± 0.4 mm). The synthesized AgNPs demonstrated excellent dye degradation kinetics in non-photocatalytic dyes methyl orange (96 %; 30 min) and safranin O (95 %; 40 min) along with experimental reusability found to be satisfactorily upto 7 cycles.
本研究首次报道了利用海草Thalassia hemprichii叶片提取物(The)合成环境友好型纳米银(AgNPs)。用几种光谱方法描述了AgNPs。agnps的紫外可见光谱研究报道了420 nm处的吸光度峰。在傅里叶变换红外光谱中可以看到本征拉伸,推断出AgNPs生物合成中存在多种功能物种及其作用。电镜和zeta电位研究证实了AgNPs的均匀形貌、球形和高稳定性。生物合成的纳米银在30 μg/ml下具有抗菌、抗糖尿病(α-淀粉酶(77%)、α-葡萄糖苷酶(78%)、抗氧化(85-90%)、抗炎(45-60%)和染料降解等性能。银纳米颗粒对病原菌金黄色葡萄球菌和大肠杆菌的抑菌效果分别为13±0.3和21±0.2 mm,与标准庆大霉素(22±0.4 mm)相当。合成的AgNPs在非光催化染料甲基橙(96%,30分钟)和红花素O(95%, 40分钟)中表现出良好的染料降解动力学,实验可重复使用7次。
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引用次数: 0
Detection method for CRISPR-Cas9 edited tomato with single nucleotide deletion using multiplex probe-based real-time PCR and confirmation of non-transgenicity: A case study 基于多重探针的实时PCR检测CRISPR-Cas9编辑番茄单核苷酸缺失及其非转基因性的方法:一个案例研究
IF 2.8 Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2025-12-01 Epub Date: 2025-10-29 DOI: 10.1016/j.jgeb.2025.100609
Monika Singh, Shiwani, Raghavendra Aminedi, Kushaldeep Kaur
The adoption of gene-edited (GE) food crops has increased due to applicability of technology without drastically affecting the genetic information. However, the discussion on regulatory regime pertaining to GE plants throughout the globe is overwhelming. In the countries where GE products are regulated, or for the verification of gene-editing as claimed by the developers, detection of GE plants (referred to as GE detection here) plays an imperative role. Precisely detecting small modifications in single or few nucleotide(s) is a challenge. An efficient detection method was developed for GE tomato line with a single base pair edit as deletion in Solanum lycopersicum pectate lyase (SlPL) gene for better shelf life. A stepwise strategy is reported herein, comprising of screening at early phase targeting Cas9 protein gene using rapid loop-mediated isothermal amplification (LAMP) and conventional polymerase chain reaction (PCR) assays, followed by the verification of single point mutation using real-time PCR. Multiplex TaqMan® real-time PCR using fluorescent labelled dual probes simultaneously targeting edited and unedited sequences, was used for verification of single point deletion. This method was based on the negative selection where the presence of a mutation was determined by absence of signal in comparison to the wild-type. The developed multiplex real-time PCR method was found sensitive enough to detect 0.1% of the targeted lines. In view of the regulation of GM tomato in several countries, the approach for confirming non-transgenic nature of edited line was also presented using real-time PCR targeting common screening elements present in globally approved GM tomato events. This approach could be utilized as a tool globally either to support the regulatory authorities for food traceability or to verify the editing as well as to track suspected transgenic element in SDN-1 and SDN-2 types.
基因编辑(GE)粮食作物的采用由于技术的适用性而不会严重影响遗传信息而增加。然而,关于全球范围内与转基因植物有关的监管制度的讨论是压倒性的。在转基因产品受到监管的国家,或者为了验证开发者声称的基因编辑,对转基因植物的检测(这里简称为GE检测)起着至关重要的作用。精确地检测单个或几个核苷酸的小修饰是一个挑战。建立了转基因番茄果胶裂解酶(SlPL)基因单碱基对编辑缺失的高效检测方法。本文报道了一种循序渐进的策略,包括使用快速环介导等温扩增(LAMP)和传统聚合酶链反应(PCR)检测在早期筛选靶向Cas9蛋白基因,然后使用实时PCR验证单点突变。多重TaqMan®实时PCR使用荧光标记双探针同时靶向编辑和未编辑的序列,用于验证单点缺失。该方法是基于负选择,其中通过与野生型相比缺乏信号来确定突变的存在。发现所开发的多重实时PCR方法灵敏度足以检测0.1%的目标系。鉴于一些国家对转基因番茄的监管,本文还提出了利用实时荧光定量PCR技术,针对全球批准的转基因番茄事件中常见的筛选元件,确定编辑系非转基因性质的方法。这种方法可以作为一种工具在全球范围内用于支持食品可追溯性的监管部门或验证编辑以及跟踪SDN-1和SDN-2类型中的可疑转基因元素。
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引用次数: 0
Prognostic and functional role of PPAR-alpha and SNP receptor (Leu162Val) in acute coronary syndrome: a potential novel target ppar - α和SNP受体(Leu162Val)在急性冠脉综合征中的预后和功能作用:一个潜在的新靶点
IF 2.8 Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2025-12-01 Epub Date: 2025-08-20 DOI: 10.1016/j.jgeb.2025.100555
Abdullah Abdulsattar Raeef , Hassan H. Al-Saeed , Sami Mekhlif Mishlish

Background and objective

PPAR-alpha plays a key role in acute coronary syndrome (ACS). Its activation influences lipid metabolism and inflammation, impacting cardiovascular health and potentially mitigating risks related to heart conditions. This study aims to find the relationship between PPAR-alpha concentration and genetic polymorphism in ACS patients.

Methods and materials

The study enrolled 90 ACS patients and 90 controls aged 30–70. Blood samples were analyzed for biomarkers (troponin, CK-MB, hs-CRP, PPARα) and rs1800206 SNP via ELISA and HRM-PCR. Data were statistically analyzed using SPSS v21, ANOVA, and ROC tests.

Results

The results showed an increase in the levels of troponin, CK-MB, and hs-CRP for the patient groups compared to the healthy groups with significant differences between the groups (p-value <0.05). The level of (PPAR-alpha) was low in patients (1472.2 ± 74.5) and high in the healthy group (2051.8 ± 149.9) with significant differences between study groups. ROC analysis identified CK-MB, and hs-CRP as optimal biomarkers for ACS diagnosis; PPARα showed good predictive value. The study also found that the dominant genotype (CC) represented a small percentage of the patient samples while the homozygotes (CG) and the recessive (GG) represented the highest percentage, unlike the healthy group. Also, Allele (C) has a low percentage in the patients’ group while allele (G) has a high percentage with considering the G allele as a risk factor. In addition, the results showed an abnormal distribution of Hardy-Weinberg equilibrium indicating the presence of factors extraneous to the community that changed the genetic pattern. The results also showed that the dominant genetic model is the best expressive for the samples of the Iraqi community infected with ACS.

Conclusions

The present study demonstrated a relationship between the concentration of PPAR-alpha and the genetic polymorphism of Leu162Val (rs1800206) SNP and acute coronary syndrome with a decrease in the concentration of PPAR-alpha and the predominance of the recessive gene and the recessive allele in patients compared to healthy individuals.
背景与目的ppar - α在急性冠脉综合征(ACS)中起关键作用。它的激活会影响脂质代谢和炎症,影响心血管健康,并可能降低与心脏病相关的风险。本研究旨在探讨ACS患者ppar - α浓度与基因多态性的关系。方法与材料本研究纳入90例ACS患者和90例30-70岁的对照组。采用ELISA和HRM-PCR检测血液中生物标志物(肌钙蛋白、CK-MB、hs-CRP、PPARα)和rs1800206 SNP。数据采用SPSS v21、方差分析和ROC检验进行统计学分析。结果患者组肌钙蛋白、CK-MB、hs-CRP水平均高于健康组,组间差异有统计学意义(p值<;0.05)。患者ppar - α水平低(1472.2±74.5),健康组ppar - α水平高(2051.8±149.9),组间差异有统计学意义。ROC分析发现CK-MB和hs-CRP是ACS诊断的最佳生物标志物;PPARα具有良好的预测价值。该研究还发现,与健康组不同,显性基因型(CC)在患者样本中所占比例很小,而纯合子(CG)和隐性基因型(GG)所占比例最高。在考虑G等位基因为危险因素时,等位基因(C)在患者组中的百分比较低,而等位基因(G)在患者组中的百分比较高。此外,Hardy-Weinberg平衡呈异常分布,表明群落外因素的存在改变了遗传模式。结果还表明,显性遗传模型在伊拉克社区感染ACS的样品中表现最好。结论ppar - α浓度与Leu162Val (rs1800206) SNP遗传多态性与急性冠状动脉综合征存在一定的关系,且ppar - α浓度降低,且隐性基因和隐性等位基因在患者中较健康人群占优势。
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引用次数: 0
Transcriptome profiling by RNA sequencing reveals novel targets of Gemini nano curcumin on p53-mutant HT-29 colorectal cancer cells RNA测序转录组分析揭示了Gemini纳米姜黄素对p53突变HT-29结直肠癌细胞的新靶点
IF 2.8 Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2025-12-01 Epub Date: 2025-08-20 DOI: 10.1016/j.jgeb.2025.100548
Hewa Jalal Azeez , Raheleh Karimzadeh , Esmaeil Babaei

Introduction

Colorectal cancer is considered as an aggressive tumor with high mortality in the world. It has been shown that Gemini Nano-Curcumin (Gemini-Cur) affects viability of colorectal cancer cells. Nevertheless, the cellular and molecular mechanisms underlying its toxicity are debatable. Here, we planned to untangle the potential novel targets for nanocurcumin on p53-mutant HT-29 cancer cells by employing RNA sequencing.

Methods

After cultivation, HT-29 cell were incubated with appropriate doses of Gemini-Cur for 24 h. Total RNA was extracted, cDNA library was constructed and sequenced. The DESeq2 tool were employed to normalize reads and detect differentially expressed genes (DEGs). The enrichR tool was employed to do Gene Ontology (GO) & identify biological processes (BP), cellular components (CC) and molecular functions (MF) that are impacted in the condition. The PPI network was constructed of 1200 DEGs using STRING, visualized by Cytoacape and analyzed with MCODE.

Results

After DEGs screening, 1309 genes between untreated and treated cancer cells were obtained including 479 upregulated with P-value < 0.05 & log2 FC > 1) as well as 63 downregulated genes with P-value < 0.05 and log2 FC < −1). Then, DEGs were assigned to 207 GO terms including numerous cellular pathways such as endoplasmic reticulum (ER)-related processes. Finally, 542 up and downregulated genes were mapped to 67 reactome pathways. The pathway analysis illustrated that Gemini-Cur modulates numerous pathways including ER stress response- and transporter-related genes. Using MCODE, Three modules were significantly identified with scores ≥ 1 and nodes ≥ 1.

Conclusion

Our data reveal that nanocurcumin might affects HT-29 colorectal cancer cells through modulating different pathways such as endoplasmic reticulum response, and transporter-related genes. More studies necessitate to unravel the molecular mechanisms and proteins involved in these pathways that could be considered as novel therapeutic targets in colorectal cancer.
结直肠癌在世界范围内被认为是一种高死亡率的侵袭性肿瘤。研究表明,Gemini纳米姜黄素(Gemini- cur)可以影响结直肠癌细胞的生存能力。然而,其毒性背后的细胞和分子机制是有争议的。在这里,我们计划通过RNA测序来解开纳米姜黄素在p53突变HT-29癌细胞上的潜在新靶点。方法HT-29细胞培养后,用适当剂量的Gemini-Cur孵育24 h,提取总RNA,构建cDNA文库并测序。采用DESeq2工具将reads归一化并检测差异表达基因(DEGs)。利用富集工具进行基因本体(GO)鉴定在该条件下受影响的生物过程(BP)、细胞成分(CC)和分子功能(MF)。用STRING构建了1200个deg的PPI网络,用cytocape进行可视化,并用MCODE进行分析。结果经DEGs筛选,在未处理和处理的癌细胞间共获得1309个基因,其中p值<; 0.05 & log2 FC < 1)上调基因479个,p值<; 0.05 & log2 FC < 1)下调基因63个。然后,deg被分配到207个氧化石墨烯术语,包括许多细胞途径,如内质网(ER)相关过程。最终,542个上调和下调基因被定位到67个反应组通路上。通路分析表明,Gemini-Cur调节多种通路,包括内质网应激反应和转运蛋白相关基因。使用MCODE,三个模块被显著识别,得分≥1,节点≥1。结论纳米姜黄素可能通过调节内质网反应、转运蛋白相关基因等不同途径影响HT-29结直肠癌细胞。需要更多的研究来揭示这些途径的分子机制和蛋白质,这些途径可能被认为是结直肠癌的新治疗靶点。
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引用次数: 0
In vitro antiproliferative effects of green synthesized silver nanoparticles from Brassica carinata microgreens on DU-145 prostate cancer cells and In vivo safety assessment 绿合成银纳米颗粒对DU-145前列腺癌细胞的体外抗增殖作用及体内安全性评价
IF 2.8 Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2025-12-01 Epub Date: 2025-09-19 DOI: 10.1016/j.jgeb.2025.100552
Dogfounianalo Somda , Joel L. Bargul , Sabina Wangui Wachira , John M. Wesonga

Background

Prostate cancer ranks among the most lethal malignancies affecting men globally. Conventional therapeutic approaches for this disease encounter limitations due to adverse side effects and the emergence of drug resistance, making its management challenging. Green-synthesized silver nanoparticles (AgNPs) have emerged as an attractive alternative to address the shortcomings of existing anticancer treatments.

Objective

This study was undertaken to evaluate the anticancer potential and safety profile of BCM-AgNPs, focusing on DU-145 inhibition and sub-acute toxicity in Wistar rats.

Methods

Previously synthesized Brassica carinata microgreen silver nanoparticles (BCM-AgNPs) alongside Brassica carinata microgreen crude extract (BCME) were used. The antiproliferative activity was investigated through the MTT assay on DU-145 prostate cancer cells and normal Vero E6 cells to determine the IC50 and CC50. Subsequently, a series of bioassays, including cell migration assays, clonogenic assays, cell cycle assays, and DNA fragmentation assays, were conducted. Furthermore, RT-qPCR was performed to delineate the molecular mechanism of BCM-AgNPs on DU-145 cells. Finally, a subacute toxicity study was carried out to assess the safety of BCM-AgNPs.

Results

Both BCM-AgNPs and BCME exhibited potent antiproliferative activity against DU-145 cells with an IC50 of 33.47 μg/mL and 46.25 μg/mL, respectively, while sparing normal Vero E6 cells with selectivity indices of 3.50 for BCM-AgNPs and 4.62 for BCME. BCM-AgNPs demonstrated anti-migratory effects and hindered colony formation in DU-145 cells. They further induced S-phase cell cycle arrest through DNA fragmentation. RT-qPCR analysis revealed the upregulation of pro-apoptotic genes and the downregulation of genes involved in cell survival and proliferation pathways. BCM-AgNPs showed no toxicity in Wistar rats during 28-day oral administration.

Conclusion

This study demonstrates for the first time the selective and prominent antiproliferative effects of BCM-AgNPs against DU-145, as well as their safety and biocompatibility, underscoring their potential as a promising candidate for anticancer therapy.
前列腺癌是影响全球男性的最致命的恶性肿瘤之一。由于不良副作用和耐药性的出现,这种疾病的传统治疗方法受到限制,使其管理具有挑战性。绿色合成的银纳米颗粒(AgNPs)已经成为解决现有抗癌治疗缺点的一个有吸引力的替代方案。目的评价BCM-AgNPs的抗癌潜力和安全性,重点研究其对DU-145的抑制作用和对Wistar大鼠的亚急性毒性。方法采用前人合成的芸苔微绿银纳米颗粒(BCM-AgNPs)和芸苔微绿粗提物(BCME)。采用MTT法测定DU-145前列腺癌细胞和正常Vero E6细胞的IC50和CC50,研究其抗增殖活性。随后,进行了一系列生物测定,包括细胞迁移测定、克隆生成测定、细胞周期测定和DNA片段化测定。此外,RT-qPCR方法进一步揭示了BCM-AgNPs作用于DU-145细胞的分子机制。最后,进行了亚急性毒性研究,以评估BCM-AgNPs的安全性。结果BCM-AgNPs和BCME对DU-145细胞均有较强的抑制活性,IC50分别为33.47 μg/mL和46.25 μg/mL,而对正常Vero E6细胞均有抑制作用,BCM-AgNPs的选择性指数为3.50,BCME的选择性指数为4.62。BCM-AgNPs在DU-145细胞中具有抗迁移作用,并能抑制集落形成。他们进一步通过DNA断裂诱导s期细胞周期阻滞。RT-qPCR分析显示,促凋亡基因上调,参与细胞存活和增殖途径的基因下调。BCM-AgNPs对Wistar大鼠口服28天无毒性。结论本研究首次证明了BCM-AgNPs对DU-145的选择性和显著的抗增殖作用,以及它们的安全性和生物相容性,强调了它们作为抗癌治疗的潜在候选药物的潜力。
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引用次数: 0
Genome-wide characterization and seasonal modulation of Hsp60 expression in juvenile Hilsa Shad across distinct upstream habitats 不同上游生境鲥鱼幼鱼Hsp60表达的全基因组特征和季节性调节
IF 2.8 Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2025-12-01 Epub Date: 2025-10-07 DOI: 10.1016/j.jgeb.2025.100586
Md.Toasin Hossain Aunkor , Afroza Pervin , Md.Nazmul Hasan , Zobada Kanak Khan , Rakibul Islam Akanda , Mohammad Mehedi Hasan Khan , Kazi Mohammad Ali Zinnah , Md.Faruque Miah
Heat shock protein 60 acts as a molecular chaperone that assists in proper protein folding under stress conditions. However, its genomic features and temperature-responsive behavior have not yet been studied in Hilsa Shad. In this study, we employed both computational and experimental approaches to investigate the genomic, proteomic, and expression dynamics of Hsp60 in this ecologically and economically important anadromous species. We identified three distinct gene copies which showed high sequence similarity compared to the Hsp60 gene of the well-studied three-spined stickleback. The physico-chemical properties, gene structure, functional domains and motifs, and sequence alignment were analyzed to characterize the paralogs. The phylogenetic analysis and structural modeling further confirmed their close evolutionary relationship and high structural similarity. However, Hilsa juvenile experience winter in the Meghna river in February and autumn in the Surma river in September due to their upstream migration. Notably, all three Hsp60 gene paralogs were significantly upregulated in the gill and liver during the warmer autumn season. While the level of relative expression was moderate in muscle and modest in kidney. This finding suggests a strong role for these genes in the thermal stress response of Hilsa Shad in the Surma river. In addition to, it demonstrates the role of Hsp60 in maintaining homeostasis in juvenile Hilsa Shad under fluctuating environmental conditions. It also aligns with the established role of heat shock proteins in thermal biology, where Hsp60 facilitates cellular protection and adaptation of juvenile Hilsa Shad to changing thermal regimes.
热休克蛋白60作为一种分子伴侣,在应激条件下帮助适当的蛋白质折叠。然而,其基因组特征和温度响应行为尚未在Hilsa Shad中研究。在这项研究中,我们采用计算和实验两种方法研究了Hsp60在这种生态和经济上重要的溯河产卵物种中的基因组学、蛋白质组学和表达动态。我们鉴定出三个不同的基因拷贝,与研究充分的三棘棘鱼的Hsp60基因序列相似。分析了其理化性质、基因结构、功能域和基序、序列比对等特征。系统发育分析和结构建模进一步证实了它们密切的进化关系和高度的结构相似性。然而,希尔萨幼崽由于上游迁徙,2月在梅克纳河过冬,9月在苏尔马河过冬。值得注意的是,在温暖的秋季,所有三种Hsp60基因在鳃和肝脏中都显着上调。而在肌肉和肾脏中相对表达水平中等。这一发现表明,这些基因在苏尔玛河Hilsa Shad的热应激反应中起着重要作用。此外,它还证明了Hsp60在波动环境条件下维持鲥鱼幼鱼体内平衡的作用。这也与热休克蛋白在热生物学中的既定作用一致,其中Hsp60促进了Hilsa Shad幼鱼对不断变化的热环境的细胞保护和适应。
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引用次数: 0
Identification of high-risk signatures and therapeutic targets through molecular characterization and immune profiling of TP53-mutant breast cancer 通过tp53突变型乳腺癌的分子特征和免疫分析鉴定高危特征和治疗靶点
IF 2.8 Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2025-12-01 Epub Date: 2025-09-30 DOI: 10.1016/j.jgeb.2025.100574
Peter Jerome Ishmael V. Paulino, Mohammad Tasyriq Che Omar

Background

TP53 mutations are commonly observed in aggressive subtypes of breast cancer, influencing the tumor microenvironment (TME) and patient prognosis. In this study, we developed a prognostic gene-based risk model to stratify TP53-mutant breast cancer patients and explore potential therapeutic targets.

Methods

We performed comprehensive bioinformatics analyses using TCGA and METABRIC datasets to identify key prognostic genes in TP53-mutant breast cancer. Differential expression and Gene Set Enrichment Analysis (GSEA) revealed dysregulated pathways, while protein–protein interaction (PPI) networks highlighted functional hubs. Survival analysis, followed by univariate Cox regression, LASSO, and multivariate regression, led to the construction of a robust gene-based risk model. Immune landscape profiling was conducted to evaluate tumor microenvironment characteristics. Finally, drug sensitivity analysis and molecular docking were used to identify potential therapeutic agents targeting high-risk patients.

Results

TP53 mutations were present in ∼ 35 % of patients and associated with significant transcriptomic alterations. A total of 666 genes were consistently dysregulated, including 333 upregulated (such as A2ML1, CA9, VGLL1, PSAT1) and 333 downregulated (such as AGR3, TFF1, ESR1, CPB1) in TP53 mutated breast cancer patients. GSEA revealed that the cell cycle, DNA replication, and metabolic pathways in in TP53 mutated breast cancer patients. Protein–protein interaction (PPI) network analysis of these genes revealed tightly connected modules related to mitotic regulation and immune signaling, underscoring key functional hubs in TP53-mutant tumors. A four-gene prognostic model (FGFR4, S100P, ADM, CTSC) stratified TP53-mutant patients into high- and low-risk groups with distinct survival outcomes and immune profiles. High-risk patients exhibited a suppressed immune landscape, characterized by lower immune and stromal cell infiltration and higher tumor purity. Drug sensitivity analysis and molecular docking revealed several compounds, including Lapatinib, Docetaxel, and Trametinib, with strong binding affinities to key model genes. These drugs demonstrated potential efficacy in high-expression cells, suggesting their viability as targeted therapies.

Conclusion

Our findings underscore the prognostic value of the identified genes and the immunosuppressive TME in TP53-mutant breast cancer. The identification of drug candidates with strong binding affinities to key proteins provides promising avenues for targeted therapy in this high-risk patient population.
背景tp53突变常见于侵袭性乳腺癌亚型,影响肿瘤微环境(TME)和患者预后。在这项研究中,我们建立了一个基于预后基因的风险模型来对tp53突变的乳腺癌患者进行分层,并探索潜在的治疗靶点。方法采用TCGA和METABRIC数据集进行综合生物信息学分析,确定tp53突变乳腺癌的关键预后基因。差异表达和基因集富集分析(GSEA)揭示了失调的途径,而蛋白质相互作用(PPI)网络突出了功能枢纽。生存分析,然后是单变量Cox回归、LASSO和多变量回归,构建了一个强大的基于基因的风险模型。采用免疫景观分析方法评价肿瘤微环境特征。最后,通过药物敏感性分析和分子对接,确定针对高危患者的潜在治疗药物。结果stp53突变存在于约35%的患者中,并与显著的转录组改变相关。共有666个基因持续失调,其中在TP53突变乳腺癌患者中有333个基因上调(如A2ML1、CA9、VGLL1、PSAT1), 333个基因下调(如AGR3、TFF1、ESR1、CPB1)。GSEA揭示了TP53突变乳腺癌患者的细胞周期、DNA复制和代谢途径。这些基因的蛋白-蛋白相互作用(PPI)网络分析揭示了与有丝分裂调节和免疫信号相关的紧密连接的模块,强调了tp53突变肿瘤的关键功能枢纽。四基因预后模型(FGFR4, S100P, ADM, CTSC)将tp53突变患者分为高风险和低风险组,具有不同的生存结果和免疫特征。高危患者表现为免疫抑制,免疫和基质细胞浸润较低,肿瘤纯度较高。药物敏感性分析和分子对接显示,包括拉帕替尼、多西他赛和曲美替尼在内的几种化合物与关键模型基因具有很强的结合亲和力。这些药物在高表达细胞中显示出潜在的疗效,表明它们作为靶向治疗的可行性。结论我们的研究结果强调了鉴定的基因和免疫抑制TME在tp53突变乳腺癌中的预后价值。与关键蛋白具有强结合亲和力的候选药物的鉴定为这一高危患者群体的靶向治疗提供了有希望的途径。
{"title":"Identification of high-risk signatures and therapeutic targets through molecular characterization and immune profiling of TP53-mutant breast cancer","authors":"Peter Jerome Ishmael V. Paulino,&nbsp;Mohammad Tasyriq Che Omar","doi":"10.1016/j.jgeb.2025.100574","DOIUrl":"10.1016/j.jgeb.2025.100574","url":null,"abstract":"<div><h3>Background</h3><div>TP53 mutations are commonly observed in aggressive subtypes of breast cancer, influencing the tumor microenvironment (TME) and patient prognosis. In this study, we developed a prognostic gene-based risk model to stratify TP53-mutant breast cancer patients and explore potential therapeutic targets.</div></div><div><h3>Methods</h3><div>We performed comprehensive bioinformatics analyses using TCGA and METABRIC datasets to identify key prognostic genes in TP53-mutant breast cancer. Differential expression and Gene Set Enrichment Analysis (GSEA) revealed dysregulated pathways, while protein–protein interaction (PPI) networks highlighted functional hubs. Survival analysis, followed by univariate Cox regression, LASSO, and multivariate regression, led to the construction of a robust gene-based risk model. Immune landscape profiling was conducted to evaluate tumor microenvironment characteristics. Finally, drug sensitivity analysis and molecular docking were used to identify potential therapeutic agents targeting high-risk patients.</div></div><div><h3>Results</h3><div>TP53 mutations were present in ∼ 35 % of patients and associated with significant transcriptomic alterations. A total of 666 genes were consistently dysregulated, including 333 upregulated (such as <em>A2ML1, CA9, VGLL1, PSAT1</em>) and 333 downregulated (such as <em>AGR3, TFF1, ESR1, CPB1</em>) in TP53 mutated breast cancer patients. GSEA revealed that the cell cycle, DNA replication, and metabolic pathways in in TP53 mutated breast cancer patients. Protein–protein interaction (PPI) network analysis of these genes revealed tightly connected modules related to mitotic regulation and immune signaling, underscoring key functional hubs in TP53-mutant tumors. A four-gene prognostic model (<em>FGFR4, S100P, ADM, CTSC</em>) stratified TP53-mutant patients into high- and low-risk groups with distinct survival outcomes and immune profiles. High-risk patients exhibited a suppressed immune landscape, characterized by lower immune and stromal cell infiltration and higher tumor purity. Drug sensitivity analysis and molecular docking revealed several compounds, including Lapatinib, Docetaxel, and Trametinib, with strong binding affinities to key model genes. These drugs demonstrated potential efficacy in high-expression cells, suggesting their viability as targeted therapies.</div></div><div><h3>Conclusion</h3><div>Our findings underscore the prognostic value of the identified genes and the immunosuppressive TME in TP53-mutant breast cancer. The identification of drug candidates with strong binding affinities to key proteins provides promising avenues for targeted therapy in this high-risk patient population.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100574"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145227466","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
Potential antibacterial, antioxidant and anticancer activities of Nigerian mountain honey in relation to its bacterial flora 尼日利亚山蜜的抗菌、抗氧化和抗癌活性与其菌群的关系
IF 2.8 Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2025-12-01 Epub Date: 2025-08-25 DOI: 10.1016/j.jgeb.2025.100559
Sara H. Mansour , Asmaa I. El-shazly , Amira A. Gamal , Elsayed A. Elsayed , Mohammad A. Wadaan , Mona A. Esawy
Numerous studies have investigated the biological activities of honey, emphasizing its antimicrobial, antioxidant, and therapeutic properties. However, a comprehensive examination of the bacteria isolated from honey has been largely overlooked. This study aimed to bridge that gap by exploring the relationship between the bioactivities of honey and the bacterial spores it contains. Furthermore, there is no comparison between the polysaccharides in honey such as levan and its inherent bacteria. Accordingly, various enzymes, including levansucrase, invertase, glucose oxidase, amylase, and catalase, were assessed in Nigerian mountain honey and their inherent seven bacterial isolates. Honey and its isolates showed levansucrase production to various degrees where the highest activity was obtained with isolate NO.4 and the honey recorded (21U/ml). Additionally, honey and its isolates showed varied activities between (171–465 U/ml), (88–250 U/ml) for amylase and glucose oxidase respectively. The highest catalase activity was recorded by isolate No 4 based on the qualitative test. Interestingly, all isolates lacked protease activity. Honey and specific isolates inhibited various pathogenic bacteria, including Salmonella enterica Typhi, Escherichia coli, and Bacillus cereus. Also, fungi such as Candida albicans. In addition, honey and its isolates had high antioxidant activities ranging from 58 to 77%. The polysaccharides Lev1, Lev2, and honey have no cytotoxic effects on normal cells. They were identified as levans based on FTIR and NMR. Honey and lev1, lev2 showed cytotoxicity against HCT116 cell lines (61.5, 59.5, and 53.70%), respectively. Interestingly, the study found a strong link between the biological activities of the bacteria and the honey itself.
许多研究调查了蜂蜜的生物活性,强调其抗菌、抗氧化和治疗特性。然而,对从蜂蜜中分离出来的细菌的全面检查在很大程度上被忽视了。这项研究旨在通过探索蜂蜜的生物活性和它所含的细菌孢子之间的关系来弥合这一差距。此外,也没有比较蜂蜜中的多糖,如levan和其固有的细菌。因此,各种酶,包括左旋蔗糖酶,转化酶,葡萄糖氧化酶,淀粉酶和过氧化氢酶,评估了尼日利亚山蜂蜜及其固有的7种细菌分离物。蜂蜜及其分离物均能产生不同程度的左旋蔗糖酶,其中分离物NO.4和蜂蜜的活性最高(21U/ml)。此外,蜂蜜及其分离物对淀粉酶和葡萄糖氧化酶的活性分别在171 ~ 465 U/ml、88 ~ 250 U/ml之间。定性试验结果显示,4号分离物过氧化氢酶活性最高。有趣的是,所有分离株都缺乏蛋白酶活性。蜂蜜和特定的分离物抑制多种致病菌,包括伤寒沙门氏菌、大肠杆菌和蜡样芽孢杆菌。此外,真菌,如白色念珠菌。此外,蜂蜜及其分离物的抗氧化活性在58% ~ 77%之间。多糖Lev1、Lev2和蜂蜜对正常细胞无细胞毒作用。经FTIR和NMR鉴定为levans。蜂蜜和level1、level2对HCT116细胞株的细胞毒性分别为61.5%、59.5%和53.70%。有趣的是,研究发现细菌的生物活性和蜂蜜本身有很强的联系。
{"title":"Potential antibacterial, antioxidant and anticancer activities of Nigerian mountain honey in relation to its bacterial flora","authors":"Sara H. Mansour ,&nbsp;Asmaa I. El-shazly ,&nbsp;Amira A. Gamal ,&nbsp;Elsayed A. Elsayed ,&nbsp;Mohammad A. Wadaan ,&nbsp;Mona A. Esawy","doi":"10.1016/j.jgeb.2025.100559","DOIUrl":"10.1016/j.jgeb.2025.100559","url":null,"abstract":"<div><div>Numerous studies have investigated the biological activities of honey, emphasizing its antimicrobial, antioxidant, and therapeutic properties. However, a comprehensive examination of the bacteria isolated from honey has been largely overlooked. This study aimed to bridge that gap by exploring the relationship between the bioactivities of honey and the bacterial spores it contains. Furthermore, there is no comparison between the polysaccharides in honey such as levan and its inherent bacteria. Accordingly, various enzymes, including levansucrase, invertase, glucose oxidase, amylase, and catalase, were assessed in Nigerian mountain honey and their inherent seven bacterial isolates. Honey and its isolates showed levansucrase production to various degrees where the highest activity was obtained with isolate NO.4 and the honey recorded (21U/ml). Additionally, honey and its isolates showed varied activities between (171–465 U/ml), (88–250 U/ml) for amylase and glucose oxidase respectively. The highest catalase activity was recorded by isolate No 4 based on the qualitative test. Interestingly, all isolates lacked protease activity. Honey and specific isolates inhibited various pathogenic bacteria, including <em>Salmonella enterica Typhi, Escherichia coli,</em> and <em>Bacillus cereus.</em> Also, fungi such as <em>Candida albicans.</em> In addition, honey and its isolates had high antioxidant activities ranging from 58 to 77%. The polysaccharides Lev1, Lev2, and honey have no cytotoxic effects on normal cells. They were identified as levans based on FTIR and NMR. Honey and lev1, lev2 showed cytotoxicity against HCT116 cell lines (61.5, 59.5, and 53.70%), respectively. Interestingly, the study found a strong link between the biological activities of the bacteria and the honey itself.</div></div>","PeriodicalId":53463,"journal":{"name":"Journal of Genetic Engineering and Biotechnology","volume":"23 4","pages":"Article 100559"},"PeriodicalIF":2.8,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144896453","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
The effects of gut microbiota-derived short-chain fatty acids (SCFAs) on human colon cancer cells: An in vitro study 肠道微生物来源的短链脂肪酸(SCFAs)对人结肠癌细胞的影响:一项体外研究
IF 2.8 Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2025-12-01 Epub Date: 2025-10-29 DOI: 10.1016/j.jgeb.2025.100610
Shiva Darabi , Fatemeh Keshavarzi , Parviz Ashtari , Farahnaz Motamedi Sedeh , Behrouz Alirezapour
Colorectal cancer (CRC) has always been a major concern for researchers due to its deadly nature. One of the microbial metabolites found in the gut is short-chain fatty acids (SCFAs), which have the ability to regulate the immune system and exert protective effects against CRC. In this study, we investigated the effects of SCFAs on the expression of the CYP1A1 gene in Caco-2 cells at varying concentrations. The Caco-2 cell line and HDF control cell line were treated with three concentrations (7.5, 15, and 30 mM) of NaA(sodium acetate), NaB(sodium butyrate), and NaP(sodium propionate) for three different time periods (24, 48, and 72 h). Additionally, two concentrations (2.5 and 10 μM) of CH223191 (an AhR gene antagonist) alone and a concentration of 10 μM of CH223191 in combination with the mentioned concentrations of NaA, NaB and NaP were applied to the cells for 48 h. The gene expression of CYP1A1 was examined using Real-time PCR. The expression of the CYP1A1 gene increased with concentrations of NaA and NaB at all time points, and with 30 mM of NaP at 24 h (p < 0.05). However, increasing the concentration of NaB and NaP led to decreased expression of the CYP1A1 gene. These findings suggest the potential use of SCFAs as epigenetic drugs for the prevention and treatment of CRC.
结直肠癌(CRC)由于其致命的性质一直是研究人员关注的主要问题。在肠道中发现的微生物代谢物之一是短链脂肪酸(SCFAs),它具有调节免疫系统并对结直肠癌发挥保护作用的能力。在这项研究中,我们研究了不同浓度的SCFAs对Caco-2细胞中CYP1A1基因表达的影响。Caco-2细胞系和HDF对照细胞系分别用3种浓度(7.5、15和30 mM)的NaA(乙酸钠)、NaB(丁酸钠)和NaP(丙酸钠)处理3个不同的时间段(24、48和72 h)。分别用2.5 μM和10 μM浓度的CH223191 (AhR基因拮抗剂)和10 μM浓度的CH223191与NaA、NaB和NaP联合作用于细胞48 h,采用Real-time PCR检测CYP1A1基因的表达情况。CYP1A1基因的表达随NaA和NaB浓度在各时间点及30mm NaP在24 h时的升高而升高(p < 0.05)。然而,NaB和NaP浓度的增加导致CYP1A1基因的表达降低。这些发现提示SCFAs作为表观遗传药物用于预防和治疗结直肠癌的潜在应用。
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引用次数: 0
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Journal of Genetic Engineering and Biotechnology
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