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Eco-friendly synthesis of betanin-conjugated zinc oxide nanoparticles: antimicrobial efficacy and apoptotic pathway activation in oral cancer cells. 以生态友好的方式合成甜菜宁共轭氧化锌纳米粒子:在口腔癌细胞中的抗菌功效和凋亡途径激活。
IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-07 DOI: 10.1007/s11033-024-10039-0
Chinnasamy Ragavendran, Mohamed Imath, Chinnaperumal Kamaraj, Ismini Nakouti, Salim Manoharadas

Background: Phytochemical-based synthesis of nanoparticles (NPs) is an eco-friendly approach with various biomedical applications. Betanin, a natural pigment in beetroot, has antioxidant, anti-inflammatory, and antimicrobial properties. When conjugated with zinc oxide nanoparticles (ZnO NPs), these properties are enhanced. This study aimed to synthesize betanin-ZnO nanoparticles (BE-ZnO-NPs) and evaluate their biological potential.

Methods: BE-ZnO-NPs were synthesized and characterized using UV-Visible spectroscopy, FTIR, FE-SEM, HR-TEM, EDX, XRD, DLS, and zeta potential analysis. In silico studies assessed interactions with oral pathogen proteins, and antibacterial activity was tested against Enterococcus faecalis, Candida albicans, Staphylococcus aureus and Streptococcus mutans. Antioxidant potential and cytotoxicity on KB cells were evaluated through scavenging assays, MTT assay, and qRT-PCR.

Results: Betanin synthesized ZnO NPs UV-Vis results showed surface plasmon resonance at 388 nm, and FTIR confirmed betanin role as a capping agent. FE-SEM and TEM revealed particles of 37 nm. EDX confirmed zinc content, and XRD showed a hexagonal structure. Zeta potential was - 3.3 mV, and DLS indicated a size of 38.73 nm. In silico analysis showed strong binding to E. faecalis (- 8.0 Kcal/mol). BE-ZnO-NPs demonstrated antibacterial activity at 100 µg/mL, with inhibition zones of 18 ± 0.14 mm for E. faecalis and 14 ± 0.18 mm for S. mutans. In contrast, BE demonstrated antibacterial activity at 100 µg/mL, with zone of inhibition of 10.6 ± 0.14 mm for E. faecalisand 11.4 ± 0.18 mm for S. mutans.Antioxidant assays revealed dose-dependent scavenging activity. Cytotoxicity showed an IC50 of 24.29 µg/mL, with qRT-PCR indicating apoptosis through the BCL2/BAX/P53 pathway.

Conclusions: BE-ZnO-NPs exhibited significant antibacterial and antioxidant activities and demonstrated the ability to induce apoptosis in oral cancer cells via the BCL-2/BAX/P53 signalling pathway. These findings highlight the potential of BE-ZnO-NPs as promising antimicrobial agents for tooth infections and as therapeutic agents for oral tumour treatment.

背景:以植物化学物质为基础合成纳米粒子(NPs)是一种生态友好型方法,具有多种生物医学应用。甜菜苷是甜菜根中的一种天然色素,具有抗氧化、抗炎和抗菌特性。当与氧化锌纳米粒子(ZnO NPs)共轭时,这些特性会得到增强。本研究旨在合成甜菜宁-氧化锌纳米粒子(BE-ZnO-NPs)并评估其生物潜力:方法:合成 BE-ZnO-NPs 并使用紫外可见光谱、傅立叶变换红外光谱、FE-SEM、HR-TEM、EDX、XRD、DLS 和 zeta 电位分析对其进行表征。硅学研究评估了与口腔病原体蛋白质的相互作用,并测试了对粪肠球菌、白色念珠菌、金黄色葡萄球菌和变异链球菌的抗菌活性。此外,还通过清除试验、MTT 试验和 qRT-PCR 试验评估了贝塔宁对 KB 细胞的抗氧化潜力和细胞毒性:结果:白桦脂素合成的 ZnO NPs 紫外可见光谱结果显示在 388 纳米波长处存在表面等离子体共振,傅立叶变换红外光谱证实了白桦脂素作为封端剂的作用。FE-SEM 和 TEM 显示颗粒大小为 37 nm。电离辐射 X 射线衍射(EDX)证实了锌的含量,X 射线衍射(XRD)显示了六边形结构。Zeta 电位为 - 3.3 mV,DLS 显示其大小为 38.73 nm。硅学分析表明,BE-ZnO-NPs 与粪肠球菌的结合力很强(- 8.0 Kcal/mol)。BE-ZnO-NPs 在 100 µg/mL 的浓度下具有抗菌活性,对粪肠球菌的抑制区为 18 ± 0.14 mm,对突变酵母菌的抑制区为 14 ± 0.18 mm。相比之下,BE 在 100 µg/mL 时具有抗菌活性,对粪大肠杆菌的抑制区为 10.6 ± 0.14 mm,对变异杆菌的抑制区为 11.4 ± 0.18 mm。细胞毒性的 IC50 值为 24.29 µg/mL,qRT-PCR 表明细胞凋亡是通过 BCL2/BAX/P53 途径进行的:BE-ZnO-NPs具有显著的抗菌和抗氧化活性,并能通过BCL-2/BAX/P53信号通路诱导口腔癌细胞凋亡。这些发现凸显了 BE-ZnO-NPs 作为治疗牙齿感染的抗菌剂和口腔肿瘤治疗剂的潜力。
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引用次数: 0
The expression of shelterin genes and telomere repeat analysis and their effect on Alzheimer's disease. 庇护素基因的表达和端粒重复分析及其对阿尔茨海默病的影响。
IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-06 DOI: 10.1007/s11033-024-10063-0
Şenay Görücü Yılmaz, Hakan Bozkurt

Background: Alzheimer's disease (AD) is an age-related dementia disorder characterized by memory loss and behavioral changes. Maintaining the integrity of telomere shortening in AD is important for cellular survival and homeostasis in all cells, especially glial cells. The shelterin protein complex provides telomere integrity. Measuring the expression levels of shelterin genes and determining the telomere lengths regulated by this complex will reveal their effects on AD progression and adult neurogenesis and will allow the detection of the disease or the determination of the progression process from an accessible tissue.

Methods and results: The study population included 111 patients and 91 healthy controls (male and female, < 50 age). The clinical histories (age, gender, hypertension, diabetes mellitus, obesity, cardiovascular disease, MMSE, medication use, family history, sleep disorders, seizure), covariates (HGB, ESR, Na, P, Cl, BUN, CRP, B12, TSH, Glucose, and MRI findings) and the expressional changes of shelterin genes (TERF1, TERF2, TINF2, POT1, TPP1, and RAP1) between the patient and control groups were evaluated relatively. ROC analyses determined the diagnostic power of telomere repeats and gene expressions.

Conclusions: In conclusion, upregulation of expression of shleterin complex genes was detected in AD, where telomeres are significantly shorter than in controls (P < 0.05). However, only TERF2 and RAP1 were significant (P < 0.05). A positive relationship was detected between telomere repeats and these genes (P < 0.05). Telomere repeats may be a strong diagnostic criterion to distinguish AD individuals from healthy individuals (AUC = 1.000). The upregulation of TERF2 and RAP1 core genes required for telomere integrity results in the instability of excessively shortened telomeres. Expression silencing of these genes may increase telomerase activity and maintain cellular survival. Also, the detection of telomere repeats has potential in the early diagnosis of AD patients.

背景:阿尔茨海默病(AD)是一种与年龄有关的痴呆症,以记忆丧失和行为改变为特征。在阿尔茨海默病中保持端粒缩短的完整性对于所有细胞,尤其是神经胶质细胞的存活和平衡非常重要。庇护蛋白复合物提供了端粒的完整性。测量保护蛋白基因的表达水平并确定由该复合物调控的端粒长度,将揭示它们对AD进展和成人神经发生的影响,并能从可触及的组织中检测疾病或确定进展过程:研究对象包括111名患者和91名健康对照者(男性和女性):总之,在AD患者中发现了shleterin复合基因的表达上调,AD患者的端粒明显短于对照组(P<0.05)。
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引用次数: 0
Exploring the interplay between DHCR7, vitamin D deficiency, and type 2 diabetes mellitus (T2DM): a systematic review. 探索 DHCR7、维生素 D 缺乏症和 2 型糖尿病 (T2DM) 之间的相互作用:系统综述。
IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-06 DOI: 10.1007/s11033-024-10072-z
Walaa Mohammedsaeed

Type 2 diabetes mellitus (T2DM) is a growing global health concern. The pathogenesis of T2DM is multifactorial and intricate, involving a complex interplay of genetic predisposition, environmental factors, and molecular interactions. Vitamin D (circulating 25-hydroxyvitamin D concentration) regulates factors crucial for T2DM, including insulin secretion, sensitivity, and inflammation. Thus, vitamin D deficiency has been linked to poor health outcomes in T2DM patients. The cholesterol-synthesizing enzyme 7-dehydrocholesterol reductase (DHCR7) represents a critical regulatory switch between cholesterol and vitamin D3 synthesis. Recent findings suggest that the enzyme DHCR7 may indicate T2DM glycolipid metabolic disorder and is associated with deficient circulating vitamin D (circulating 25-hydroxyvitamin D concentration) status. In this PRISMA-guided systematic review, articles were sourced from two databases, namely, PubMed and Cochrane Library, to evaluate the impact of vitamin D deficiency in patients with T2DM and to explore the emerging role of DHCR7 in T2DM pathogenesis. Our findings strongly indicate a positive correlation between deficient vitamin D status and poor health outcomes in T2DM patients. Finally, this systematic review presents a novel perspective on T2DM development, focusing on the interplay between T2DM-associated hyperglycemia, expression of DHCR7, and abrogation of vitamin D synthesis.

2 型糖尿病(T2DM)是全球日益关注的健康问题。T2DM 的发病机制是多因素的,错综复杂,涉及遗传易感性、环境因素和分子相互作用的复杂相互作用。维生素 D(循环中 25- 羟基维生素 D 的浓度)调节 T2DM 的关键因素,包括胰岛素分泌、敏感性和炎症。因此,维生素 D 缺乏与 T2DM 患者的不良健康状况有关。胆固醇合成酶 7-脱氢胆固醇还原酶(DHCR7)是胆固醇和维生素 D3 合成之间的关键调节开关。最近的研究结果表明,DHCR7 酶可能预示着 T2DM 糖脂代谢紊乱,并与循环维生素 D(循环 25- 羟维生素 D 浓度)缺乏状态有关。在这篇以 PRISMA 为指导的系统性综述中,我们从 PubMed 和 Cochrane Library 两个数据库中收集了文章,以评估维生素 D 缺乏对 T2DM 患者的影响,并探讨 DHCR7 在 T2DM 发病机制中的新作用。我们的研究结果强烈表明,维生素 D 缺乏与 T2DM 患者的不良健康后果之间存在正相关。最后,这篇系统综述从一个新的视角探讨了 T2DM 的发展,重点是 T2DM 相关高血糖、DHCR7 的表达和维生素 D 合成障碍之间的相互作用。
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引用次数: 0
Fluorescent primers amplification refractory mutation system qPCR (FP ARMS-qPCR) for MTHFR C677T SNP genotyping. 用于 MTHFR C677T SNP 基因分型的荧光引物扩增难治突变系统 qPCR (FP ARMS-qPCR)。
IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-06 DOI: 10.1007/s11033-024-10036-3
Yongjun Han, Hong Wu

Background: The currently used methods for the detection of methylene tetrahydrofolic acid reductase (MTHFR) C677T single nucleotide polymorphism (SNP) are either time-consuming or expensive. In this study, we devised an accurate, rapid and easy-to-use SNP detection system based on fluorescent primers amplification refractory mutation system qPCR, known as FP ARMS-qPCR.

Methods: Fluorescent primers (FPs) modified by fluorescent dye or quencher near the 3' terminal thymine were designed. The reaction conditions were optimized and the performance was evaluated. Using commercial kits as controls, 242 samples were tested in parallel to verify the feasibility of the FP ARMS-qPCR assay.

Results: We demonstrated the good sensitivity and specificity of the FP ARMS-qPCR with optimized conditions. The assay was able to accurately distinguish between different SNP sites of MTHFR C677T in less than 2 h using as low as 50 pg of template genomic DNA. Completely consistent genotyping results reveal that FP ARMS-qPCR is concordant with commercial kits.

Conclusion: We established a specific, sensitive, and rapid FP ARMS-qPCR method for the detection of MTHFR C677T genotype. This could also serve as a potential diagnostic tool for a variety of diseases.

背景:目前用于检测亚甲基四氢叶酸还原酶(MTHFR)C677T单核苷酸多态性(SNP)的方法要么费时,要么昂贵。在这项研究中,我们设计了一种基于荧光引物扩增难治性突变系统 qPCR 的准确、快速、易用的 SNP 检测系统,即 FP ARMS-qPCR:方法:设计了在 3' 末端胸腺嘧啶附近用荧光染料或淬灭剂修饰的荧光引物(FPs)。对反应条件进行了优化,并对性能进行了评估。以商业试剂盒为对照,对 242 份样本进行了平行测试,以验证 FP ARMS-qPCR 分析的可行性:结果:在优化条件下,我们证明了 FP ARMS-qPCR 具有良好的灵敏度和特异性。在不到 2 小时的时间内,该检测方法就能准确区分 MTHFR C677T 的不同 SNP 位点,使用的模板基因组 DNA 低至 50 pg。完全一致的基因分型结果表明,FP ARMS-qPCR 与商业试剂盒的结果一致:我们建立了一种特异、灵敏、快速的 FP ARMS-qPCR 方法来检测 MTHFR C677T 基因型。结论:我们建立了特异、灵敏、快速的 FP ARMS-qPCR 方法来检测 MTHFR C677T 基因型,这也可作为多种疾病的潜在诊断工具。
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引用次数: 0
Phylogenetic and population genetic analysis of Arunachali Yak (Bos grunniens) using mitochondrial DNA D-loop region. 利用线粒体 DNA D 环区对阿鲁纳恰里牦牛(Bos grunniens)进行系统发育和种群遗传分析。
IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-06 DOI: 10.1007/s11033-024-10076-9
Martina Pukhrambam, Atrayee Dutta, Pranab Jyoti Das, Anesha Chanda, Mihir Sarkar

Background: The Arunachali yak (Bos grunniens) is an important breed native to the northeastern Himalayas of India. Understanding its genetic diversity and evolutionary relationships with other bovine species is crucial for effective conservation and breeding strategies. This study investigates the mitochondrial DNA (mtDNA) D-loop region of Arunachali yak and compares it with other bovines to elucidate these relationships.

Methods: We collected blood samples from 18 Arunachali yak individuals and isolated genomic DNA. The partial mtDNA D-loop region was amplified using PCR and sequenced. Sequences were compared with those from Bos frontalis, Bos gaurus, Bos indicus, Bubalus bubalis, and Capra hircus available in GenBank. Phylogenetic relationships were assessed through Neighbor-Joining trees and median-joining networks. Genetic diversity indices and neutrality tests were applied to evaluate population genetic characteristics.

Results: Phylogenetic analysis identified three distinct clades, with Arunachali yak clustering closely with Bos indicus, and forming a separate branch from other bovine species. Median-joining networks revealed six haplogroups, with Arunachali yak uniquely representing Hap_3. Genetic diversity analysis showed no polymorphism within Arunachali yak, indicating very low genetic variation in the selected animal samples. AMOVA demonstrated significant genetic differentiation among populations (Fst = 0.30053, P < 0.001), with a substantial portion of variation occurring within populations.

Conclusions: The Arunachali yak exhibits a close genetic relationship with Bos indicus, reflecting recent divergence. The study underscores the distinct genetic profile of Arunachali yak and highlights its limited genetic variability. These findings enhance our understanding of bovine evolutionary relationships and emphasize the need for targeted conservation measures to preserve the genetic integrity of Arunachali yak.

背景:阿鲁纳恰里牦牛(Bos grunniens)是原产于印度喜马拉雅山脉东北部的一个重要品种。了解阿鲁纳恰里牦牛的遗传多样性以及与其他牛类的进化关系对于有效的保护和育种策略至关重要。本研究调查了阿鲁纳恰里牦牛的线粒体 DNA(mtDNA)D 环区,并将其与其他牛类进行比较,以阐明这些关系:我们采集了 18 头阿鲁纳恰里牦牛的血液样本,并分离了基因组 DNA。方法:我们采集了 18 头阿鲁纳恰里牦牛的血液样本,并分离了基因组 DNA,使用 PCR 扩增了部分 mtDNA D-loop 区域并进行了测序。序列与 GenBank 中的 Bos frontalis、Bos gaurus、Bos indicus、Bubalus bubalis 和 Capra hircus 序列进行了比较。通过邻接树和中位连接网络评估了系统发育关系。应用遗传多样性指数和中性检验来评估种群遗传特征:系统发育分析发现了三个不同的支系,阿鲁纳恰里牦牛与牛密切聚类,并与其他牛类形成一个独立的分支。中位连接网络显示了六个单倍群,阿鲁纳恰里牦牛是 Hap_3 的唯一代表。遗传多样性分析表明,阿鲁纳恰里牦牛内部没有多态性,这表明所选动物样本的遗传变异非常小。AMOVA表明种群间存在明显的遗传差异(Fst = 0.30053,P 结论):阿鲁纳恰里牦牛与印度牦牛的遗传关系密切,反映了最近的分化。这项研究强调了阿鲁纳恰里牦牛独特的遗传特征,并突出了其有限的遗传变异性。这些发现加深了我们对牛进化关系的理解,并强调了采取有针对性的保护措施以保护阿鲁纳恰里牦牛遗传完整性的必要性。
{"title":"Phylogenetic and population genetic analysis of Arunachali Yak (Bos grunniens) using mitochondrial DNA D-loop region.","authors":"Martina Pukhrambam, Atrayee Dutta, Pranab Jyoti Das, Anesha Chanda, Mihir Sarkar","doi":"10.1007/s11033-024-10076-9","DOIUrl":"https://doi.org/10.1007/s11033-024-10076-9","url":null,"abstract":"<p><strong>Background: </strong>The Arunachali yak (Bos grunniens) is an important breed native to the northeastern Himalayas of India. Understanding its genetic diversity and evolutionary relationships with other bovine species is crucial for effective conservation and breeding strategies. This study investigates the mitochondrial DNA (mtDNA) D-loop region of Arunachali yak and compares it with other bovines to elucidate these relationships.</p><p><strong>Methods: </strong>We collected blood samples from 18 Arunachali yak individuals and isolated genomic DNA. The partial mtDNA D-loop region was amplified using PCR and sequenced. Sequences were compared with those from Bos frontalis, Bos gaurus, Bos indicus, Bubalus bubalis, and Capra hircus available in GenBank. Phylogenetic relationships were assessed through Neighbor-Joining trees and median-joining networks. Genetic diversity indices and neutrality tests were applied to evaluate population genetic characteristics.</p><p><strong>Results: </strong>Phylogenetic analysis identified three distinct clades, with Arunachali yak clustering closely with Bos indicus, and forming a separate branch from other bovine species. Median-joining networks revealed six haplogroups, with Arunachali yak uniquely representing Hap_3. Genetic diversity analysis showed no polymorphism within Arunachali yak, indicating very low genetic variation in the selected animal samples. AMOVA demonstrated significant genetic differentiation among populations (Fst = 0.30053, P < 0.001), with a substantial portion of variation occurring within populations.</p><p><strong>Conclusions: </strong>The Arunachali yak exhibits a close genetic relationship with Bos indicus, reflecting recent divergence. The study underscores the distinct genetic profile of Arunachali yak and highlights its limited genetic variability. These findings enhance our understanding of bovine evolutionary relationships and emphasize the need for targeted conservation measures to preserve the genetic integrity of Arunachali yak.</p>","PeriodicalId":18755,"journal":{"name":"Molecular Biology Reports","volume":"51 1","pages":"1125"},"PeriodicalIF":2.6,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142583384","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Genome-wide investigation of MYB gene family in Areca catechu and potential roles of AcTDF in transgenic Arabidopsis. 儿茶属 MYB 基因家族的全基因组调查以及 AcTDF 在转基因拟南芥中的潜在作用。
IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-05 DOI: 10.1007/s11033-024-10055-0
Qiyuan An, Yiqi Jiang, Guangzhen Zhou

Background: MYB protein, a crucial transcription factor, holds crucial importance in plant growth, development, stress responses, and secondary metabolite regulation. While MYB proteins have been extensively studied, research on MYBs within the palm family, particularly in Areca catechu, remains limited.

Methods and results: This study identified 259 MYB genes in Areca catechu, including 105 1R-MYBs, 150 R2R3-MYBs, 3 3R-MYBs, and 1 4R-MYBs. Physicochemical properties, collinearity, and gene structure of these genes were analyzed. The AcMYB is distributed across 16 chromosomes of A.catechu and has 119 and 195 homologs in Arabidopsis and rice, respectively. Cis-acting elements in the promoter region suggest roles in plant hormones, growth, development, and stress. R2R3-MYB genes were divided into eight groups based on tissue expression profiles. The flowering-related gene AcTDF is highly expressed in male flowers. Overexpression of AcTDF in Arabidopsis promotes early flowering, upregulates AtSOC1 and AtFUL, and enhances tolerance to drought and salt stress.

Conclusions: These results provide valuable insights for the identification and analysis of the MYB gene family in Areca catechu and offer a basis for the subsequent verification of its related functions and the role and significance of its role in the evolution of palms.

背景:MYB 蛋白是一种重要的转录因子,在植物的生长、发育、胁迫反应和次生代谢物调控中具有至关重要的作用。虽然对 MYB 蛋白进行了广泛的研究,但对棕榈科植物,尤其是对儿茶属植物中的 MYB 的研究仍然有限:这项研究在儿茶属植物中发现了 259 个 MYB 基因,包括 105 个 1R-MYB、150 个 R2R3-MYB、3 个 3R-MYB 和 1 个 4R-MYB。对这些基因的理化性质、共线性和基因结构进行了分析。AcMYB 分布在 A.catechu 的 16 条染色体上,在拟南芥和水稻中分别有 119 和 195 个同源基因。启动子区域的顺式作用元件表明其在植物激素、生长、发育和胁迫中发挥作用。根据组织表达谱将 R2R3-MYB 基因分为八组。与开花相关的基因 AcTDF 在雄花中高表达。在拟南芥中过表达 AcTDF 可促进早开花,上调 AtSOC1 和 AtFUL,并增强对干旱和盐胁迫的耐受性:这些结果为鉴定和分析辣木籽中的 MYB 基因家族提供了有价值的见解,并为后续验证其相关功能及其在棕榈树进化中的作用和意义提供了依据。
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引用次数: 0
Branched-chain amino acids and their metabolites decrease human and rat hepatic stellate cell activation. 支链氨基酸及其代谢物可降低人和大鼠肝星状细胞的活化程度
IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-04 DOI: 10.1007/s11033-024-10027-4
Maria Camila Trillos-Almanza, Magnolia Martinez Aguilar, Manon Buist-Homan, Nils Bomer, Karla Arevalo Gomez, Vincent E de Meijer, Frederike G I van Vilsteren, Hans Blokzijl, Han Moshage

Background: End-stage liver diseases (ESLDs) are a significant global health challenge due to their high prevalence and severe health impacts. Despite the severe outcomes associated with ESLDs, therapeutic options remain limited. Targeting the activation of hepatic stellate cells (HSCs), key drivers of extracellular matrix accumulation during liver injury presents a novel therapeutic approach. In ESLDs patients, branched-chain amino acids (BCAAs, leucine, isoleucine and valine) levels are decreased, and supplementation has been proposed to attenuate liver fibrosis and improve regeneration. However, their effects on HSCs require further investigation.

Objective: To evaluate the efficacy of BCAAs and their metabolites, branched-chain α-keto acids (BCKAs), in modulating HSCs activation in human and rat models.

Methods: Primary HSCs from rats and cirrhotic and non-cirrhotic human livers, were cultured and treated with BCAAs or BCKAs to assess their effects on both preventing (from day 1 of isolation) and reversing (from day 7 of isolation) HSCs activation.

Results: In rat HSCs, leucine and BCKAs significantly reduced fibrotic markers and cell proliferation. In human HSCs, the metabolite of isoleucine decreased cell proliferation around 85% and increased the expression of branched-chain ketoacid dehydrogenase. The other metabolites also showed antifibrotic effects in HSCs from non-cirrhotic human livers.

Conclusion: BCAAs and their respective metabolites inhibit HSC activation with species-specific responses. Further research is needed to understand how BCAAs influence liver fibrogenesis. BCKAs supplementation could be a strategic approach for managing ESLDs, considering the nutritional status and amino acid profiles of patients.

背景:终末期肝病(ESLDs)发病率高,对健康影响严重,是全球健康面临的重大挑战。尽管 ESLDs 会导致严重后果,但治疗方案仍然有限。肝星状细胞是肝损伤过程中细胞外基质积聚的关键驱动因素,针对肝星状细胞的活化提出了一种新的治疗方法。在 ESLDs 患者中,支链氨基酸(BCAAs,亮氨酸、异亮氨酸和缬氨酸)水平降低,补充支链氨基酸可减轻肝纤维化并改善再生。然而,它们对造血干细胞的影响还需要进一步研究:目的:评估 BCAAs 及其代谢产物支链 α-酮酸(BCKAs)在人类和大鼠模型中调节造血干细胞活化的功效:方法:用支链α-酮酸或支链α-酮酸培养和处理来自大鼠、肝硬化和非肝硬化人类肝脏的原发性造血干细胞,以评估它们对预防(从分离第1天起)和逆转(从分离第7天起)造血干细胞活化的影响:结果:在大鼠造血干细胞中,亮氨酸和碱性磷酸酶能显著减少纤维化标志物和细胞增殖。在人造血干细胞中,异亮氨酸代谢物可使细胞增殖减少约 85%,并增加支链酮酸脱氢酶的表达。其他代谢物在非肝硬化的人类肝脏造血干细胞中也显示出抗纤维化作用:结论:BCAAs 及其各自的代谢物可抑制造血干细胞的活化,并具有物种特异性反应。要了解 BCAAs 如何影响肝纤维化,还需要进一步的研究。考虑到患者的营养状况和氨基酸谱,补充 BCKAs 可能是治疗 ESLD 的一种战略方法。
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引用次数: 0
PPM1D/Wip1 is amplified, overexpressed, and mutated in human non-Hodgkin's lymphomas. PPM1D/Wip1在人类非霍奇金淋巴瘤中扩增、过表达和突变。
IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-04 DOI: 10.1007/s11033-024-10029-2
Hatice Pilevneli, Firuzan Döger, Levent Karagenç, Didem Kozacı, Mehtap Kilic Eren

Background: Wip1, is a p53-dependent Ser/Thr phosphatase involved in the timely termination of DDR. The PPM1D gene encoding Wip1 is deregulated and thus gained an oncogene character in common human solid tumors and cell lines. This study assessed the oncogenic potential of the PPM1D gene in human non- Hodgkin's lymphomas (NHL), the most common hematological malignancy worldwide.

Methods and results: FFPE human lymphoid hyperplasia (LH) (n = 17) and NHL tumor lymph node samples (n = 65) and human NHL cell lines were used to assess the oncogenic potential of the PPM1D gene in the present study. Copy number gain and mRNA expression analysis of the PPM1D/Wip1 gene were assessed by qRT-PCR analysis. Mutational analysis of Exon 6 of the PPM1D gene was performed by PCR amplification and Sanger sequencing. Expressions of Wip1 and p53 proteins were assessed by immunohistochemistry and Western blot analysis.

Conclusions: We found that PPM1D gained gene copy number in NHL tumors by 0.7-8 times compared to the control (p < 0.01). Increased PPM1D/Wip1 gene copy number was associated with higher mRNA and protein expression in human NHL samples (p < 0.01). Overexpression of Wip1 in NHL tumors and NHL cell lines was associated with amplification level and was unaffected by p53 status. Furthermore, a heterozygous type insertion mutation was detected in exon 6 (c.1553_1554insA) of the PPM1D gene particularly in DLBCL samples. Wip1 may have oncogenic potential, perhaps playing a role in the onset and progression of human NHL. The possible significance of Wip1 overexpression to chemotherapy response in NHL remains an intriguing question that requires more exploration.

背景Wip1 是一种依赖于 p53 的 Ser/Thr 磷酸酶,参与 DDR 的及时终止。在常见的人类实体瘤和细胞系中,编码 Wip1 的 PPM1D 基因会发生失调,从而具有致癌基因的特征。本研究评估了 PPM1D 基因在人类非霍奇金淋巴瘤(NHL)(全球最常见的血液恶性肿瘤)中的致癌潜力:本研究采用FFPE人类淋巴增生(LH)(n = 17)和NHL肿瘤淋巴结样本(n = 65)以及人类NHL细胞系来评估PPM1D基因的致癌潜力。通过 qRT-PCR 分析评估了 PPM1D/Wip1 基因的拷贝数增殖和 mRNA 表达分析。通过 PCR 扩增和 Sanger 测序对 PPM1D 基因的第 6 外显子进行了突变分析。免疫组化和 Western 印迹分析评估了 Wip1 和 p53 蛋白的表达:我们发现,与对照组相比,PPM1D 在 NHL 肿瘤中的基因拷贝数增加了 0.7-8 倍(p
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引用次数: 0
Selection and validation of reference genes for quantitative expression analysis of regeneration-related genes in Cheilomenes sexmaculata by real-time qRT-PCR. 利用实时 qRT-PCR 技术选择和验证用于雌花鲈再生相关基因定量表达分析的参考基因。
IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-04 DOI: 10.1007/s11033-024-10062-1
Shivali Pandita, Hera Alam, Radha Shivhare, Manisha Singh, Sanchita Singh, Geetanjali Mishra, Praveen C Verma

Background: Regeneration is a fascinating phenomenon that has intrigued scientists for a long time. Cheilomenes sexmaculata (Fabricius), a zig-zag ladybird beetle, possesses a high capacity for limb regeneration. The molecular mechanics of the zig-zag ladybird beetle are under-explored. Current research trends are focused on uncovering functional genes associated with limb regeneration. Most of these investigations involve quantitative real-time PCR (qRT-PCR) for their rapid and accurate analysis of gene expression levels. Hence, a stable and suitable reference gene is required to normalize the gene expression data.

Methods and results: In this study, five housekeeping genes were selected from the transcriptomics data (in-house unpublished data) of C. sexmaculata (Fabricius). The expression stabilities of the selected genes were evaluated under different time intervals post-amputation using geNorm, normFinder, and refFinder software. Actin was revealed to be the most stable housekeeping gene, along with elongation factor 2 and glyceraldehyde-3-phosphate dehydrogenase. A target gene named engrailed (an important segment-forming gene) was used to validate the selected reference genes. The expression levels were found to be consistent with the transcriptomics results.

Conclusion: According to our study, actin, along with elongation factor 2 and glyceraldehyde-3-phosphate dehydrogenase, serve as the most stable reference genes and are suitable for regeneration-related research. This study is a groundbreaking effort to identify the most stable reference gene for limb regeneration in C. sexmaculata (Fabricius), and the findings can be applied to other related insect species.

背景介绍再生是一种迷人的现象,长期以来一直吸引着科学家。Cheilomenes sexmaculata (Fabricius) 是一种人字形瓢虫,具有很强的肢体再生能力。人字形瓢虫的分子机理尚未得到充分探索。目前的研究趋势主要集中在发现与肢体再生相关的功能基因上。这些研究大多采用实时定量 PCR(qRT-PCR)技术,以快速准确地分析基因表达水平。因此,需要一个稳定且合适的参考基因来归一化基因表达数据:本研究从 C. sexmaculata(Fabricius)的转录组学数据(内部未发表数据)中选取了五个管家基因。使用 geNorm、normFinder 和 refFinder 软件评估了所选基因在截肢后不同时间间隔内的表达稳定性。结果显示,肌动蛋白是最稳定的管家基因,此外还有伸长因子 2 和甘油醛-3-磷酸脱氢酶。一个名为 engrailed 的目标基因(一个重要的片段形成基因)被用来验证所选的参考基因。研究发现,这些基因的表达水平与转录组学结果一致:根据我们的研究,肌动蛋白、伸长因子 2 和甘油醛-3-磷酸脱氢酶是最稳定的参考基因,适合再生相关研究。这项研究为确定蛙肢体再生最稳定的参考基因做出了开创性的努力,研究结果可应用于其他相关昆虫物种。
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引用次数: 0
Correction: Exome sequencing identified mutations in the WNT1 and COL1A2 genes in osteogenesis imperfecta cases. 更正:外显子组测序在成骨不全症病例中发现了 WNT1 和 COL1A2 基因突变。
IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-04 DOI: 10.1007/s11033-024-10023-8
Poonam Mehta, Rahul Vishvkarma, Sushil Gupta, Naibedya Chattopadhyay, Singh Rajender
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引用次数: 0
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Molecular Biology Reports
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