Biochemical and biophysical research communications最新文献_第5页

Trametes versicolor laccase-derived silver nanoparticles: Green synthesis, structural characterization and multifunctional biological properties 银纳米粒子：绿色合成、结构表征和多功能生物特性。

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical and biophysical research communications

Pub Date : 2024-11-14 DOI: 10.1016/j.bbrc.2024.150995

Hamed Barabadi , Melika Kamali , Kamyar Jounaki , Kimiya Karami , Salar Sadeghian-Abadi , Reza Jahani , Omid Hosseini , Salimeh Amidi

Isolated enzymes serve as advantageous platforms for the fabrication of nanomaterials. The objective of this study was to fabricate silver nanoparticles (AgNPs) incorporated with Trametes versicolor laccase and evaluate their diverse biological properties. The AgNPs fabricated through laccase-mediated methods were characterized using various characterization techniques including UV–visible (UV–vis) spectroscopy, Energy-dispersive X-ray (EDX) spectroscopy, Dynamic light scattering (DLS) spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, and Field emission scanning electron microscopy (FE-SEM). The results showed that the laccase-incorporated AgNPs were spherical in shape with a Z-average diameter of 19.40 nm and a zeta potential of −19.2 mV. The AgNPs exhibited significant dose-dependent in vitro α-amylase, urease, and DPPH free radical inhibitory activities, with maximum inhibitions of 83.49 ± 1.06 %, 68.95 ± 3.60 %, and 67.36 ± 3.40 %, respectively, at a concentration of 1000 μg mL⁻¹. Furthermore, the intrinsic pathway-mediated anticoagulant activity of the fabricated AgNPs was confirmed through the activated partial thromboplastin time (aPTT) assay, which serves as a global coagulation assay. Additionally, the laccase-incorporated AgNPs demonstrated antibacterial properties against both standard gram-positive strains of Staphylococcus epidermidis and Streptococcus mutans, with minimum inhibitory concentration (MIC) values of 2 and 4 μg mL⁻¹, and minimum bactericidal concentration (MBC) values of 16 and 16 μg mL⁻¹, respectively. The dose-dependent antibacterial performance of the AgNPs against both bacterial populations was also confirmed through flow cytometry. Moreover, the AgNPs exhibited 61.53 ± 3.17 % and 63.03 ± 1.44 % biofilm degradation against S. epidermidis and S. mutans, respectively, at the maximum tested concentration (20∗MIC).

分离酶是制造纳米材料的有利平台。本研究的目的是制备掺入了Trametes versicolor漆酶的银纳米粒子（AgNPs），并评估其各种生物特性。研究采用了多种表征技术，包括紫外可见光谱（UV-vis）、能量色散 X 射线（EDX）光谱、动态光散射（DLS）光谱、傅立叶变换红外光谱（FT-IR）和场发射扫描电子显微镜（FE-SEM），对通过漆酶介导的方法制备的 AgNPs 进行了表征。结果表明，加入漆酶的 AgNPs 呈球形，Z 平均直径为 19.40 nm，zeta 电位为 -19.2 mV。AgNPs 具有显著的剂量依赖性体外α-淀粉酶、脲酶和 DPPH 自由基抑制活性，在浓度为 1000 μg mL-1 时，最大抑制率分别为 83.49 ± 1.06 %、68.95 ± 3.60 % 和 67.36 ± 3.40 %。此外，通过活化部分凝血活酶时间（aPTT）检测，证实了所制备 AgNPs 的内在途径介导的抗凝活性。此外，加入漆酶的 AgNPs 对标准革兰氏阳性菌株表皮葡萄球菌和变异链球菌都具有抗菌特性，最低抑菌浓度 (MIC) 值分别为 2 和 4 μg mL-1，最低杀菌浓度 (MBC) 值分别为 16 和 16 μg mL-1。流式细胞术也证实了 AgNPs 对这两种细菌的抗菌性能具有剂量依赖性。此外，在最大测试浓度（20∗MIC）下，AgNPs 对表皮葡萄球菌和变异葡萄球菌的生物膜降解率分别为 61.53 ± 3.17 % 和 63.03 ± 1.44 %。

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引用次数: 0

16S rRNA gene sequencing for bacterial identification and infectious disease diagnosis 16S rRNA 基因测序用于细菌鉴定和传染病诊断。

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical and biophysical research communications

Pub Date : 2024-11-13 DOI: 10.1016/j.bbrc.2024.150974

Mei-Na Li , Qiang Han , Nan Wang , Ting Wang , Xue-Ming You , Shuai Zhang , Cui-Cui Zhang , Yong-Qiang Shi , Pei-Zhuang Qiao , Cheng-Lian Man , Teng Feng , Yue-Yue Li , Zhuang Zhu , Ke-Ji Quan , Teng-Lin Xu , George Fei Zhang

16S rRNA gene sequence is the most common housekeeping genetic marker to study bacterial phylogeny and taxonomy. Therefore, 16S rRNA gene sequencing has the potential to identify novel bacteria and diagnose bacteria. This study compared 16S rRNA gene sequencing with conventional PCR for bacterial identification and disease diagnosis. The bacterial community in healthy and diseased hosts was analyzed by 16S rRNA gene sequencing. 16S rRNA gene sequencing is more sensitive than conventional PCR in detecting bacteria. Moreover, 16S rRNA gene sequencing is adequate to identify novel bacteria. 16S rRNA gene sequencing demonstrated that most pathogenic bacteria persist in diseased or healthy hosts in different abundance. Pathogenic bacteria, such as well-known chicken pathogen Avibacterium paragallinarum, Ornithobacterium rhinotracheale, and Gallibacterium anatis, were identified as indicator species of diseased samples. Alpha diversity analysis showed that the healthy group species is significantly higher than in the diseased groups. Beta diversity analysis also demonstrated differences between healthy and infected groups. The study concluded that 16S rRNA gene sequencing is a more sensitive method for detecting pathogens, and microbiota analysis can distinguish between healthy and diseased samples. Eventually, 16S rRNA gene sequencing has represented the potential in human and animal clinical diagnosis and novel bacterial identification.

16S rRNA 基因序列是研究细菌系统发育和分类最常用的看家遗传标记。因此，16S rRNA 基因测序具有鉴定新型细菌和诊断细菌的潜力。本研究比较了 16S rRNA 基因测序与传统 PCR 在细菌鉴定和疾病诊断中的应用。通过 16S rRNA 基因测序分析了健康和患病宿主体内的细菌群落。在检测细菌方面，16S rRNA 基因测序比传统 PCR 更灵敏。此外，16S rRNA 基因测序还能识别新型细菌。16S rRNA 基因测序表明，大多数病原菌以不同的数量存在于患病或健康的宿主体内。病原菌被鉴定为疾病样本的指示种，如众所周知的鸡病原菌Avibacterium paragallinarum、Ornithobacterium rhinotracheale和Gallibacterium anatis。阿尔法多样性分析表明，健康组的物种明显多于患病组。Beta 多样性分析也显示了健康组和感染组之间的差异。研究认为，16S rRNA 基因测序是检测病原体的一种更灵敏的方法，微生物群分析可以区分健康样本和患病样本。最终，16S rRNA 基因测序在人类和动物的临床诊断和新型细菌鉴定方面具有巨大潜力。

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引用次数: 0

Protective effect of metformin on the NG-nitro-l-arginine methyl ester (l-NAME)-induced rat models of preeclampsia 二甲双胍对 NG-硝基-精氨酸甲酯（l-NAME）诱导的子痫前期大鼠模型的保护作用。

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical and biophysical research communications

Pub Date : 2024-11-13 DOI: 10.1016/j.bbrc.2024.150996

Huiniu Hao, Feng Li, Fang Wang, Jia Ran, Yinmin Chen, Lijun Yang, Huijing Ma, Jianli Wang, Hailan Yang

Background

Preeclampsia (PE) is a complex multi-organ disorder characterized by systemic inflammation, endothelial dysfunction, and vasoconstriction, which manifests as hypertension, with or without proteinuria. Effective preventive strategies for PE are currently lacking in clinical practice, leading to significant morbidity and mortality among mothers and newborns.

Objective

This study aims to investigate the impact of metformin (MET) on the l-NAME-induced PE rat model, focusing on the mechanisms through which MET may exert its effects.

Methods

Thirty pregnant Sprague-Dawley (SD) rats were randomly assigned to three groups: Control, PE, and PE + MET, on gestational day 0 (GD0). Regularly measure blood pressure and 24-h proteinuria, and collect tissue samples on GD20. Enzyme-linked immunosorbent assay (ELISA) was used to analyze inflammatory factors, endothelial function biomarkers, angiogenic factors, and apoptosis-related factors in the rat plasma. Western blot, RT-qPCR, and immunohistochemistry techniques were employed to determine the expression levels of key apoptotic proteins in placental tissue.

Results

The study findings demonstrate that MET administration improves blood pressure and 24-h proteinuria, alleviates fetal growth restriction, ameliorate inflammation cytokines, and restores the balance of angiogenic factors and endothelial function. Moreover, MET inhibits the expression levels of critical apoptotic proteins in the plasma and placental tissue of PE-like rats.

Conclusion

The results suggest that MET shows promise in alleviating symptoms associated with l-NAME-induced PE in rats, preserving endothelial function, enhancing angiogenesis, reducing inflammation, inhibiting placental apoptosis, improving placental function, and promoting fetal growth. These findings highlight MET as a potential therapeutic agent for the prevention and treatment of preeclampsia.

背景：子痫前期（PE）是一种复杂的多器官疾病，以全身炎症、内皮功能障碍和血管收缩为特征，表现为高血压，伴有或不伴有蛋白尿。目前，临床上缺乏有效的 PE 预防策略，导致母亲和新生儿的发病率和死亡率显著上升：本研究旨在探讨二甲双胍（MET）对 l-NAME 诱导的 PE 大鼠模型的影响，重点是二甲双胍发挥其作用的机制：方法：将 30 只妊娠 Sprague-Dawley (SD) 大鼠随机分为三组，分别为对照组、PE 组和 PE + MET 组：方法：30 只妊娠 Sprague-Dawley (SD) 大鼠在妊娠第 0 天（GD0）被随机分为三组：对照组、PE 组和 PE + MET 组。定期测量血压和 24 小时蛋白尿，并在 GD20 时采集组织样本。使用酶联免疫吸附试验（ELISA）分析大鼠血浆中的炎症因子、内皮功能生物标志物、血管生成因子和细胞凋亡相关因子。研究采用了 Western 印迹、RT-qPCR 和免疫组化技术来确定胎盘组织中关键凋亡蛋白的表达水平：研究结果表明，服用 MET 可改善血压和 24 小时蛋白尿，缓解胎儿生长受限，改善炎症细胞因子，恢复血管生成因子的平衡和内皮功能。此外，MET还能抑制PE样大鼠血浆和胎盘组织中关键凋亡蛋白的表达水平：结果表明，MET有望缓解l-NAME诱导的PE大鼠的相关症状，保护内皮功能，增强血管生成，减轻炎症反应，抑制胎盘凋亡，改善胎盘功能，促进胎儿生长。这些研究结果突出表明，MET 是预防和治疗子痫前期的一种潜在治疗药物。

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引用次数: 0

Influence of anti-fibrillation TNGQ peptide and rutin combination on β-cell cytoprotective effects against IAPP-induced cell death and oxidative stress 抗纤颤TNGQ肽和芦丁组合对β细胞抗IAPP诱导的细胞死亡和氧化应激的细胞保护作用的影响

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical and biophysical research communications

Pub Date : 2024-11-13 DOI: 10.1016/j.bbrc.2024.150976

Raliat O. Abioye , Oluwasemilogo H. Adetula , Julia Diem Hum , Chibuike C. Udenigwe

Type 2 diabetes development has been associated with islet amyloid polypeptide (IAPP) fibrillation. IAPP fibrils have various deleterious effects, such as oxidative stress and disruption of cellular membrane integrity, resulting in pancreatic β-cell toxicity. Rutin, a plant polyphenol, possesses promising cytoprotective effects as a fibrillation inhibitor. Similarly, bioactive peptides have been identified as potential inhibitors to IAPP fibrillation. In this study, the effect of peptide/polyphenol mixtures consisting of rutin and each peptide, TNGQ, MANT, and YMSV, on anti-fibrillation activity and cellular response was elucidated. Results indicated a 54.7–75.1 % decrease in thioflavin T fluorescence, confirming anti-fibrillation activity. The combination decreased the average particle diameters of IAPP more than the single inhibitors, suggesting a combined effect of peptide/rutin mixtures in enhancing anti-fibrillation activity. IAPP fibrillation-induced rat insulinoma RIN-m cell death was minimized in the presence of the peptide/rutin mixture, but the activity was lower relative to rutin alone, suggesting a non-additive effect of the mixtures. Transmission electron microscopy showed a near-complete inhibition of IAPP fibrillation by TNGQ/rutin mixtures, which translated to a decreased production of membrane-bound IAPP oligomers in RIN-m cells based on immunofluorescence staining. Additionally, TNGQ/rutin mixtures significantly decreased reactive oxygen species production by 30 %, higher than the effects of single inhibitors, but no effect was observed on glucose-stimulated insulin secretion. The results demonstrate the potential of multifunctional compounds as dual inhibitor systems in controlling IAPP fibrillation and provide insight into the implications of peptide/polyphenol mixtures towards the rational development of novel anti-diabetic nutraceutical combinations.

2 型糖尿病的发生与胰岛淀粉样多肽（IAPP）纤维化有关。IAPP 纤维具有多种有害影响，如氧化应激和破坏细胞膜完整性，从而导致胰岛β细胞中毒。芦丁是一种植物多酚，作为一种纤维化抑制剂，具有良好的细胞保护作用。同样，生物活性肽也被认为是 IAPP 纤维化的潜在抑制剂。本研究阐明了由芦丁和每种肽（TNGQ、MANT 和 YMSV）组成的肽/多酚混合物对抗纤化活性和细胞反应的影响。结果表明，硫黄素 T 荧光减少了 54.7-75.1%，证实了抗纤颤活性。与单一抑制剂相比，肽/芦丁混合物能更有效地降低 IAPP 的平均颗粒直径，这表明肽/芦丁混合物在增强抗纤颤活性方面具有联合效应。在多肽/芦丁混合物存在的情况下，IAPP纤维化诱导的大鼠胰岛素瘤RIN-m细胞死亡最小化，但活性相对于单独使用芦丁更低，这表明混合物具有非叠加效应。透射电子显微镜显示，TNGQ/芦丁混合物几乎完全抑制了IAPP的纤化，根据免疫荧光染色结果，这意味着RIN-m细胞中与膜结合的IAPP寡聚体生成减少。此外，TNGQ/芦丁混合物还能显著减少活性氧的产生，减少幅度为30%，高于单一抑制剂的效果，但对葡萄糖刺激的胰岛素分泌没有影响。研究结果证明了多功能化合物作为双重抑制剂系统在控制 IAPP 纤维化方面的潜力，并深入探讨了肽/多酚混合物对合理开发新型抗糖尿病营养保健品组合的意义。

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引用次数: 0

Ligustrazine nanoparticles inhibits epithelial-mesenchymal transition and alleviates postoperative abdominal adhesion 利格列汀纳米粒子可抑制上皮-间充质转化，减轻术后腹腔粘连。

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical and biophysical research communications

Pub Date : 2024-11-13 DOI: 10.1016/j.bbrc.2024.150994

Shasha Liang , Yifei Qian , Ying Liu , Yahui Wang , Lianlin Su , Shuai Yan

Following abdominal surgery, the occurrence of postoperative abdominal adhesion (PAA) is highly prevalent and stands out as one of the most frequently encountered complications. The effect and molecular mechanisms of Ligustrazine nanoparticles (LN) underlying epithelial-mesenchymal transition (EMT) in PAA still remain elusive.

Adhesions were induced in Male Sprague-Dawley rats by injuring the cecum (cecal abrasion model), followed by administration of LN and hyaluronate acid (HA). The mechanism was further verified by enzyme-linked immunosorbent assay, wound healing assay, si-RNA and Western blot. Animal experiments revealed that LN effectively ameliorated adhesions, notably decreased tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-8, and fibrosis, and reduced the expression of TGF-β1 and EMT related markers (Fibronectin and E-cadherin). Furthermore, in vitro experiments demonstrated that LN might inhibit the TGF-β1 FOXC2 pathway through suppressing the expression of Fibronectin, P120, and E-cadherin and ameliorating peritoneal adhesion. Collectively, our findings indicate that LN inhibits PAA formation by reducing inflammation, decreasing EMT and promoting peritoneal mesothelial cell repair. Therefore, LN might be considered a potential candidate for the treatment of PPA. However, further clinical studies are required to approve the effectiveness of LN.

腹部手术后，术后腹腔粘连（PAA）的发生率很高，是最常见的并发症之一。藁本嗪纳米颗粒（LN）在上皮-间质转化（EMT）中的作用和分子机制仍未确定。通过损伤盲肠（盲肠磨损模型）诱导雄性 Sprague-Dawley 大鼠发生粘连，然后给予 LN 和透明质酸（HA）。酶联免疫吸附试验、伤口愈合试验、si-RNA 和 Western 印迹进一步验证了这一机制。动物实验表明，LN 能有效改善粘连，显著降低肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、IL-8 和纤维化，减少 TGF-β1 和 EMT 相关标记物（纤连蛋白和 E-cadherin）的表达。此外，体外实验表明，LN可通过抑制纤连蛋白、P120和E-adherin的表达以及改善腹膜粘附性来抑制TGF-β1 FOXC2通路。总之，我们的研究结果表明，LN 可通过减轻炎症、减少 EMT 和促进腹膜间皮细胞修复来抑制 PAA 的形成。因此，LN 可被视为治疗 PAA 的潜在候选药物。不过，LN 的有效性还需要进一步的临床研究来验证。

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引用次数: 0

Glycogen metabolism in methanogens: A key pathway for metabolic response to nutrient availability 甲烷菌的糖原代谢：对养分供应做出代谢反应的关键途径

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical and biophysical research communications

Pub Date : 2024-11-13 DOI: 10.1016/j.bbrc.2024.150978

Felipe Gonzalez-Ordenes, Nicolás Herrera-Soto, Sebastián M. Muñoz, Gabriel Vallejos-Baccelliere, Sixto M. Herrera, Ignacio Aravena-Valenzuela, Andres Urrutia-Santana, Victor Castro-Fernandez, Victoria Guixé

Methanogens, which are found exclusively in the Archaea domain of life, have the potential to help solve future energy challenges by producing methane. As a result, their metabolism has attracted significant attention in recent years. Despite being unable to grow on sugars, they store glycogen, which raises intriguing questions about the role of this polymer in methanogen metabolism and the signals that trigger its degradation when methanogenic substrates are not available.

Here, we examined genomic databases to identify the enzymes responsible for glycogen synthesis and degradation in methanogens and explored the critical role of glycogen when nutrients and methanogenic substrates are scarce. Additionally, we analyzed the metabolic pathways involved in glycogen metabolism and their connection to the various types of methanogenesis exhibited by these organisms. Potential regulatory steps are proposed based on the reported effectors. Also, by employing the Alphafold3 server, the structural location of these sites in the enzyme structure was predicted, highlighting the advantages and limitations of this tool. Analysis of the allosteric effectors involved in this regulation suggests that energy charge may be the signal that triggers the metabolic switch from gluconeogenesis and glycogen storage to glycolysis and methanogenesis.

甲烷菌（Methanogens）只存在于古生菌（Archaea）中，有可能通过生产甲烷来帮助解决未来的能源挑战。因此，它们的新陈代谢近年来备受关注。尽管它们不能依靠糖类生长，但它们会储存糖原，这就提出了一些耐人寻味的问题：这种聚合物在甲烷发生器代谢中的作用，以及当甲烷底物不可用时触发其降解的信号。在这里，我们研究了基因组数据库，以确定甲烷菌中负责糖原合成和降解的酶，并探讨了糖原在营养物质和产甲烷底物缺乏时的关键作用。此外，我们还分析了糖原代谢所涉及的代谢途径及其与这些生物表现出的各种类型的甲烷生成之间的联系。根据报告的效应物，提出了潜在的调控步骤。此外，通过使用 Alphafold3 服务器，我们预测了这些位点在酶结构中的位置，突出了这一工具的优势和局限性。对参与这种调控的异生效应器的分析表明，能量电荷可能是触发代谢从糖元生成和糖原储存向糖酵解和甲烷生成转换的信号。

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引用次数: 0

Entinostat treatment causes hypophosphatemia and hypocalcemia by increasing Fgf23 in mice 恩替诺特治疗可通过增加 Fgf23 导致小鼠低磷血症和低钙血症。

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical and biophysical research communications

Pub Date : 2024-11-13 DOI: 10.1016/j.bbrc.2024.150970

Wenguang Liu , Manyu Zhang , Lili Wu , Toshihisa Komori , Haoyunyan Jin , Huilin Yang , Qing Jiang , Xin Qin

Entinostat, a class I HDACs-selective inhibitor, is currently in clinical trials for treating cancers. In some of the trials, Entinostat treatment frequently causes hypophosphatemia and/or hypocalcemia. Moreover, the effect of Entinostat treatment on bone remains incompletely understood. In this study, we found that Entinostat treatment mildly increased the trabecular but not cortical bone volume, without compromising the bone strength, the numbers of Runx2-positive cells and TRAP-positive cells, and the serum levels of P1NP and TRAP-5b. Entinostat treatment significantly reduced the level of Runx2 mRNA but not Runx2 protein, and as a trend attenuated Ctsk expression. Furthermore, Entinostat treatment did not enhance MC3T3-E1 cell proliferation in vitro. These findings suggest that Entinostat increases trabecular bone volume not by regulating osteoblastogenesis or osteoclastogenesis, but possibly by attenuating the resorption capacity. Unexpectedly, Entinostat treatment increased the expression of Fgf23, whose protein is a hormone that regulates the serum level of phosphate (Pi). Meanwhile, Entinostat treatment increased the serum level of the active form (intact) Fgf23 and reduced that of Pi and calcium (Ca) as well. This study raised a concern about the anabolic effects of Entinostat in bone, and demonstrated that Entinostat treatment causes hypophosphatemia and hypocalcemia by upregulating Fgf23 mRNA and increasing intact Fgf23 protein in serum.

恩替诺特是一种 I 类 HDACs 选择性抑制剂，目前正在进行治疗癌症的临床试验。在一些试验中，恩替诺特治疗经常导致低磷酸盐血症和/或低钙血症。此外，人们对恩替诺斯他治疗对骨骼的影响仍不完全了解。在这项研究中，我们发现恩替诺特治疗可轻度增加骨小梁的骨量，但不能增加皮质骨的骨量，同时不影响骨强度、Runx2阳性细胞和TRAP阳性细胞的数量以及P1NP和TRAP-5b的血清水平。恩替诺特治疗可明显降低Runx2 mRNA水平，但不会降低Runx2蛋白水平，并有减少Ctsk表达的趋势。此外，恩替诺特处理不会增强 MC3T3-E1 细胞的体外增殖。这些研究结果表明，恩替诺特不是通过调节成骨细胞生成或破骨细胞生成来增加骨小梁体积，而可能是通过减弱吸收能力来增加骨小梁体积。意想不到的是，恩替诺特治疗增加了 Fgf23 的表达，而 Fgf23 蛋白是一种调节血清磷酸盐（Pi）水平的激素。同时，恩替诺特治疗增加了血清中活性形式（完整的）Fgf23的水平，并降低了磷酸盐和钙（Ca）的水平。这项研究引起了人们对恩替诺特在骨骼中合成代谢作用的关注，并证明恩替诺特治疗会通过上调 Fgf23 mRNA 和增加血清中的完整 Fgf23 蛋白，导致低磷血症和低钙血症。

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引用次数: 0

Properties of phosphoramide benzoazole oligonucleotides (PABAOs). II. Structure and hybridization efficiency of N-benzoxazole derivatives 磷酰胺苯并唑寡核苷酸（PABAOs）的特性。II.N-苯并恶唑衍生物的结构和杂交效率。

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical and biophysical research communications

Pub Date : 2024-11-13 DOI: 10.1016/j.bbrc.2024.150997

Ivan I. Yushin, Victor M. Golyshev, Alina I. Novgorodtseva, Alexander A. Lomzov

New phosphate-modified nucleic acid derivatives are of great significance in basic research and biomedical applications. We have recently developed a new class of phosphoramide benzoazole oligonucleotides (PABAOs). In this work, th properties of N-benzoxazole oligodeoxyribonucleotides have been thoroughly examined. We have demonstrated the convenient automated solid-phase synthesis of oligomers with a different number of modifications (up to six) at various positions. Optical properties, thermal stability, structure, and dynamics of PABAO/DNA complexes have been investigated. The N-benzoxazole-modified phosphate group is uncharged at neutral pH and has a pKa value of 8.52. Structural analysis performed by the CD spectroscopy and MD simulation indicate B-form of PABAO/DNA duplexes. The thermal stability of PABAO/DNA complexes bearing N-benzoxazole is reduced by 2.5–6.2° per modification compared to native duplexes at standard and near physiological buffer conditions. The performed study underlines a great potential of phosphoramide benzoazole oligonucleotides for basic research, applied sciences, and biotechnology.

新的磷酸修饰核酸衍生物在基础研究和生物医学应用中具有重要意义。我们最近开发了一类新的磷酰胺苯并噁唑寡核苷酸（PABAOs）。在这项工作中，我们对 N-苯并噁唑寡核苷酸的性质进行了深入研究。我们展示了在不同位置进行不同数量（最多六次）修饰的寡聚体的便捷自动化固相合成。我们对 PABAO/DNA 复合物的光学特性、热稳定性、结构和动力学进行了研究。经 N-苯并恶唑修饰的磷酸基团在中性 pH 值下不带电，pKa 值为 8.52。通过 CD 光谱和 MD 模拟进行的结构分析表明 PABAO/DNA 双链体为 B 型。在标准和接近生理的缓冲条件下，含有 N-苯并恶唑的 PABAO/DNA 复合物的热稳定性比原生双链体每改性一次降低 2.5-6.2° 。这项研究强调了磷酰胺苯并噁唑寡核苷酸在基础研究、应用科学和生物技术方面的巨大潜力。

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引用次数: 0

Quercetin exhibits cytotoxicity in cancer cells by inducing two-ended DNA double-strand breaks 槲皮素通过诱导双端 DNA 双链断裂对癌细胞产生细胞毒性。

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical and biophysical research communications

Pub Date : 2024-11-13 DOI: 10.1016/j.bbrc.2024.150977

Yuduki Someya , Shinta Saito , Shigeki Takeda , Noritaka Adachi , Aya Kurosawa

Quercetin, a flavonoid, is involved in the induction of DNA double-strand breaks (DSBs), in addition to its antioxidant properties. Although DNA topoisomerase II (Top2) and reactive oxygen species (ROS) have been suggested as possible mechanisms through which quercetin induces DSBs, the exact mechanism remains unclear. In this study, we examined the mechanism of DSB induction by quercetin and its repair using HeLa cells and gene-knockout cell lines generated from human Nalm-6 cells. Immunofluorescence staining for γH2AX, a DSB marker, and analysis of the frequency of random integration of foreign DNA, which correlates with the number of DSBs and DSB repair pathways, indicated that quercetin induces DSBs in a concentration-dependent manner. The sensitivity assay suggested that the factor involved in quercetin-induced DSBs was not Top2. However, ROS was found to accumulate transiently in quercetin-treated HeLa cells. Furthermore, the addition of ascorbic acid increased the survival of quercetin-treated HeLa cells, suggesting that quercetin induces a transient accumulation of ROS, which in turn induces DSBs. The resulting DSBs were repaired primarily by non-homologous end-joining and homologous recombination, similar to X-ray-induced DSBs. Taken together, quercetin, used as a radiomimetic agent, has the potential to produce effects equivalent to those of an X ray-dose at a relatively low risk.

槲皮素是一种黄酮类化合物，除了具有抗氧化特性外，还参与诱导DNA双链断裂（DSB）。尽管DNA拓扑异构酶II（Top2）和活性氧（ROS）被认为是槲皮素诱导DSB的可能机制，但其确切机制仍不清楚。在这项研究中，我们使用 HeLa 细胞和由人类 Nalm-6 细胞产生的基因敲除细胞系研究了槲皮素诱导 DSB 及其修复的机制。DSB标记物γH2AX的免疫荧光染色以及与DSB数量和DSB修复途径相关的外来DNA随机整合频率分析表明，槲皮素以浓度依赖性方式诱导DSB。灵敏度试验表明，参与槲皮素诱导 DSB 的因子不是 Top2，但在槲皮素处理过的 HeLa 细胞中发现 ROS 短暂积累。此外，抗坏血酸的加入提高了槲皮素处理的 HeLa 细胞的存活率，这表明槲皮素诱导了 ROS 的短暂积累，进而诱导了 DSB。产生的DSB主要通过非同源末端连接和同源重组进行修复，这与X射线诱导的DSB相似。综上所述，槲皮素作为一种放射模拟剂，有可能在风险相对较低的情况下产生相当于X射线剂量的效应。

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引用次数: 0

Gb3 trisaccharide-bearing exosomes as a novel neutralizer for Shiga toxin type 1 含 Gb3 三糖的外泌体是 1 型志贺毒素的新型中和剂。

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical and biophysical research communications

Pub Date : 2024-11-12 DOI: 10.1016/j.bbrc.2024.150975

Krzysztof Mikołajczyk

Shiga toxin types 1 (Stx1) and 2 (Stx2), produced by Shiga toxin-producing Escherichia coli (STEC) and Shigella dysenteriae, are key virulence factors responsible for severe foodborne diseases, such as hemorrhagic colitis and hemolytic uremic syndrome (HUS). The receptors for Stxs are Gb3 and P1 glycotope, which contain the Galα1→4Gal epitope and are synthesized by human α1,4-galactosyltransferase (A4galt). Stx-related infections pose a global public health challenge, owing to the limited therapeutic options due to the restricted use of antibiotics. Therefore, there is an urgent need to develop novel therapeutic strategies. This study proposes an innovative strategy utilizing exosomes derived from CHO-Lec2 cells, which were modified with Functional-Spacer-Lipid (FSL) conjugates bearing the Gb3 carbohydrate epitope (exo-Gb3-FSL). Flow cytometry analysis confirmed the presence of Galα1→4Gal disaccharides on exo-Gb3-FSL constructs, enabling them to bind Stx1. Moreover, using CHO-Lec2 cells evaluated the ability of exo-Gb3-FSL agents to bind Stx1 and protect these cells from Stx1-mediated cytotoxicity. For Stx1-treated CHO-Lec2 cells, increased cell survival was observed when using 25 μM exo-Gb3-FSL constructs, compared to control cells. These findings highlight the potential of exosome-based anti-Stx1 agents as promising alternatives to conventional therapies. This innovative strategy may provide novel directions for studies on Stx1 neutralization, offering a valuable strategy for the treatment of Stx-related diseases.

由产志贺毒素大肠杆菌（STEC）和志贺氏痢疾杆菌产生的 1 型（Stx1）和 2 型（Stx2）志贺毒素是导致出血性结肠炎和溶血性尿毒症（HUS）等严重食源性疾病的关键致病因子。Stxs 的受体是 Gb3 和 P1 糖基，它们含有 Galα1→4Gal 表位，由人类 α1,4-半乳糖基转移酶（A4galt）合成。由于抗生素的使用受到限制，治疗方案有限，因此与 Stx 相关的感染对全球公共卫生构成了挑战。因此，迫切需要开发新的治疗策略。本研究提出了一种创新策略，利用从 CHO-Lec2 细胞中提取的外泌体，并用带有 Gb3 碳水化合物表位的功能性垫片-脂质（FSL）共轭物（exo-Gb3-FSL）对其进行修饰。流式细胞仪分析证实，外-Gb3-FSL 构建体上存在 Galα1→4Gal 二糖，使其能够结合 Stx1。此外，利用 CHO-Lec2 细胞评估了外显子-Gb3-FSL 药剂结合 Stx1 并保护这些细胞免受 Stx1 介导的细胞毒性的能力。对于经 Stx1 处理的 CHO-Lec2 细胞，与对照细胞相比，使用 25 μM 的外显子-Gb3-FSL 构建物可提高细胞存活率。这些发现凸显了基于外泌体的抗Stx1药物作为传统疗法替代品的潜力。这种创新策略可能会为 Stx1 中和研究提供新的方向，为治疗 Stx 相关疾病提供有价值的策略。

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