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Systematic design and evaluation of aptamers for VEGF and PlGF biomarkers of Preeclampsia. 系统设计和评估子痫前期血管内皮生长因子和 PlGF 生物标记物的适配体。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-27 DOI: 10.1186/s12896-024-00891-0
Samavath Mallawarachchi, Rümeysa E Cebecioglu, Majed Althumayri, Levent Beker, Sandun Fernando, Hatice Ceylan Koydemir

Preeclampsia is a potentially life-threatening condition for both mother and baby, characterized by hypertension and potential organ damage. Early diagnosis is crucial to mitigate its adverse health effects. Traditional diagnostic methods, which focus on late-manifesting symptoms like hypertension and proteinuria, underscore the need for molecular diagnostic approaches for timely detection. This study successfully designs and evaluates novel aptamers with high specificity and affinity for Vascular Endothelial Growth Factor (VEGF) and Placental Growth Factor (PlGF), biomarkers closely associated with preeclampsia. Using molecular docking, molecular dynamics simulations, and BioLayer Interferometry (BLI), we identified aptamers that demonstrated strong binding affinities, comparable or superior to traditional antibodies. Our findings suggest that these aptamers have the potential to be integrated into cost-effective, point-of-care diagnostic tools, significantly improving early detection and intervention strategies for preeclampsia. The robust performance of these aptamers marks a pivotal step toward the development of more reliable and accessible diagnostic solutions, with implications for better maternal and fetal health outcomes.

子痫前期是一种可能危及母婴生命的疾病,其特点是高血压和潜在的器官损伤。早期诊断对减轻其对健康的不利影响至关重要。传统的诊断方法主要针对高血压和蛋白尿等晚期症状,因此需要分子诊断方法来及时发现。这项研究成功地设计并评估了对血管内皮生长因子(VEGF)和胎盘生长因子(PlGF)具有高特异性和亲和力的新型适配体,这两种生长因子是与子痫前期密切相关的生物标志物。通过分子对接、分子动力学模拟和生物层干涉测量法(BLI),我们鉴定出了具有强大结合亲和力的适配体,其结合亲和力可与传统抗体相媲美甚至更胜一筹。我们的研究结果表明,这些适配体有可能被整合到具有成本效益的床旁诊断工具中,从而大大改善子痫前期的早期检测和干预策略。这些适配体的强大性能标志着我们朝着开发更可靠、更易获得的诊断解决方案迈出了关键一步,这对改善孕产妇和胎儿的健康状况具有重要意义。
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引用次数: 0
Unraveling the impact of pH, sodium concentration, and medium osmolality on Vibrio natriegens in batch processes. 揭示批处理过程中 pH 值、钠浓度和培养基渗透压对纳氏弧菌的影响。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-23 DOI: 10.1186/s12896-024-00897-8
Eva Forsten, Steffen Gerdes, René Petri, Jochen Büchs, Jørgen Magnus

Background: Vibrio natriegens, a halophilic marine γ-proteobacterium, holds immense biotechnological potential due to its remarkably short generation time of under ten minutes. However, the highest growth rates have been primarily observed on complex media, which often suffer from batch-to-batch variability affecting process stability and performance. Consistent bioprocesses necessitate the use of chemically defined media, which are usually optimized for fermenters with pH and dissolved oxygen tension (DOT) regulation, both of which are not applied during early-stage cultivations in shake flasks or microtiter plates. Existing studies on V. natriegens' growth on mineral media report partially conflicting results, and a comprehensive study examining the combined effects of pH buffering, sodium concentration, and medium osmolality is lacking.

Results: This study evaluates the influence of sodium concentration, pH buffering, and medium osmolality on the growth of V. natriegens under unregulated small-scale conditions. The maximum growth rate, time of glucose depletion, as well as the onset of stationary phase were observed through online-monitoring the oxygen transfer rate. The results revealed optimal growth conditions at an initial pH of 8.0 with a minimum of 300 mM MOPS buffer for media containing 20 g/L glucose or 180 mM MOPS for media with 10 g/L glucose. Optimal sodium chloride supplementation was found to be between 7.5 and 15 g/L, lower than previously reported ranges. This is advantageous for reducing industrial corrosion issues. Additionally, an osmolality range of 1 to 1.6 Osmol/kg was determined to be optimal for growth. Under these optimized conditions, V. natriegens achieved a growth rate of 1.97 ± 0.13 1/h over a period of 1 h at 37 °C, the highest reported rate for this organism on a mineral medium.

Conclusion: This study provides guidelines for cultivating V. natriegens in early-stage laboratory settings without pH and DOT regulation. The findings suggest a lower optimal sodium chloride range than previously reported and establish an osmolality window for optimal growth, thereby advancing the understanding of V. natriegens' physiology. In addition, this study offers a foundation for future research into the effects of different ions and carbon sources on V. natriegens.

背景:纳氏弧菌(Vibrio natriegens)是一种嗜卤海洋γ-蛋白细菌,由于其生成时间极短,不到十分钟,因此具有巨大的生物技术潜力。然而,最高的生长率主要是在复杂的培养基上观察到的,而这种培养基往往会因批次间的差异而影响工艺的稳定性和性能。稳定的生物工艺需要使用化学定义的培养基,这些培养基通常是为具有 pH 值和溶解氧张力(DOT)调节功能的发酵罐而优化的,而这两种功能在摇瓶或微孔板的早期培养过程中并不适用。关于 V. natriegens 在矿物培养基上生长的现有研究报告的结果部分相互矛盾,目前还缺乏对 pH 缓冲、钠浓度和培养基渗透压综合影响的全面研究:结果:本研究评估了钠浓度、pH 缓冲和培养基渗透压对 V. natriegens 在不受限制的小规模条件下生长的影响。通过在线监测氧转移率,观察了最大生长速率、葡萄糖耗尽时间以及静止期的开始。结果表明,最佳生长条件为初始 pH 值为 8.0,含 20 克/升葡萄糖的培养基至少需要 300 毫摩尔 MOPS 缓冲液,含 10 克/升葡萄糖的培养基至少需要 180 毫摩尔 MOPS 缓冲液。最佳氯化钠补充量为 7.5 至 15 克/升,低于之前报道的范围。这有利于减少工业腐蚀问题。此外,1 至 1.6 Osmol/kg 的渗透压范围被确定为最佳生长条件。在这些优化条件下,V. natriegens 在 37 °C、1 小时内的生长速度达到了 1.97 ± 0.13 1/h,这是该生物在矿物培养基上的最高生长速度:本研究为在没有 pH 值和 DOT 调节的早期实验室环境中培养 V. natriegens 提供了指导。研究结果表明,最佳氯化钠范围比以前报道的要低,并建立了一个最佳生长的渗透压窗口,从而加深了人们对 V. natriegens 生理学的了解。此外,这项研究还为今后研究不同离子和碳源对纳氏酵母菌的影响奠定了基础。
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引用次数: 0
Anti-inflammatory potential of aspergillus unguis SP51-EGY: TLR4-dependent effects & chemical diversity via Q-TOF LC-HRMS unguis SP51-EGY 的抗炎潜力:通过 Q-TOF LC-HRMS 分析 TLR4 依赖性效应和化学多样性
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-18 DOI: 10.1186/s12896-024-00890-1
Soad Nasr, Abdelhameed S. Dawood, Amal Mosad Ibrahim, Mohamed S. Abdel-Aziz, Walid Fayad, Anwar Abdelnaser, Faten K. Abd EL-Hady
Inflammation serves as an intricate defense mechanism for tissue repair. However, overactivation of TLR4-mediated inflammation by lipopolysaccharide (LPS) can lead to detrimental outcomes such as sepsis, acute lung injury, and chronic inflammation, often associated with cancer and autoimmune diseases. This study delves into the anti-inflammatory properties of “Aspergillus unguis isolate SP51-EGY” on LPS-stimulated RAW 264.7 macrophages. Through real-time qPCR, we assessed the expression levels of pivotal inflammatory genes, including iNOS, COX-2, TNF-α, and IL-6. Remarkably, our fungal extracts significantly diminished NO production and showed noteworthy reductions in the mRNA expression levels of the aforementioned genes. Furthermore, while Nrf2 is typically associated with modulating inflammatory responses, our findings indicate that the anti-inflammatory effects of our extracts are not Nrf2-dependent. Moreover, the chemical diversity of the potent extract (B Sh F) was elucidated using Q-TOF LC-HRMS, identifying 54 compounds, some of which played vital roles in suppressing inflammation. Most notably, compounds like granisetron, fenofibrate, and umbelliprenin were found to downregulate TNF-α, IL-1β, and IL-6 through the NF-κB signaling pathway. In conclusion, “Aspergillus unguis isolate SP51-EGY”, isolated from the Red Sea, Egypt, has been unveiled as a promising TLR4 inhibitor with significant anti-inflammatory potentials, presenting novel insights for their potential therapeutic use in inflammation.
炎症是一种复杂的组织修复防御机制。然而,脂多糖(LPS)过度激活 TLR4 介导的炎症可导致败血症、急性肺损伤和慢性炎症等有害结果,而慢性炎症通常与癌症和自身免疫性疾病相关。本研究探讨了" Unguis 曲霉分离物 SP51-EGY "对 LPS 刺激的 RAW 264.7 巨噬细胞的抗炎特性。通过实时 qPCR,我们评估了关键炎症基因的表达水平,包括 iNOS、COX-2、TNF-α 和 IL-6。值得注意的是,我们的真菌提取物明显减少了 NO 的产生,并显著降低了上述基因的 mRNA 表达水平。此外,虽然 Nrf2 通常与调节炎症反应有关,但我们的研究结果表明,我们提取物的抗炎作用并不依赖于 Nrf2。此外,我们还利用 Q-TOF LC-HRMS 对强效提取物(B Sh F)的化学多样性进行了阐释,鉴定出 54 种化合物,其中一些在抑制炎症方面发挥了重要作用。最值得注意的是,研究发现格拉司琼、非诺贝特和脐橙素等化合物可通过 NF-κB 信号通路下调 TNF-α、IL-1β 和 IL-6。总之,从埃及红海分离出的 "unguis曲霉分离物SP51-EGY "是一种很有前途的TLR4抑制剂,具有显著的抗炎潜力,为其在炎症中的潜在治疗用途提供了新的见解。
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引用次数: 0
Synthesis and evaluation of nanosized aluminum MOF encapsulating Umbelliferon: assessing antioxidant, anti-inflammatory, and wound healing potential in an earthworm model 包覆伞形酮的纳米铝 MOF 的合成与评估:在蚯蚓模型中评估抗氧化、抗炎和伤口愈合潜力
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-15 DOI: 10.1186/s12896-024-00889-8
Rabab M. Thabit, Fatma El-Zahraa A. Abd El-Aziz, A. Abu El-Fadl, A. A. Abu-Sehly, Ahmed M. Sayed
Nanoporous aluminum metal–organic framework (Al-MOF) was synthesized via solvothermal methods and employed as a carrier matrix for in vitro drug delivery of Umbelliferon (Um). The encapsulated Um was gradually released over seven days at 37 °C, using simulated body fluid phosphate-buffered saline (PBS) at pH 7.4 as the release medium. The drug release profile suggests the potential of Al-MOF nanoparticles as effective drug delivery carriers. Structural and chemical analyses of Um-loaded Al-MOF nanoparticles (Um-Al MOF) were conducted using Fourier-transform infrared (FTIR) spectroscopy, X-ray diffractometry (XRD), and ultraviolet–visible (UV–Vis) spectroscopy. Thermal gravimetric analysis (TGA) was employed to investigate the thermal stability of the Al-MOF nanoparticles, while Transmission Electron Microscopy (TEM) was utilized to assess their morphological features. Um-Al MOF nanoparticles demonstrated notable antioxidant and anti-inflammatory properties compared to Um and Al-MOF nanoparticles individually. Moreover, they exhibited significant enhancement in wound healing in an earthworm model. These findings underscore the potential of Al-MOF nanoparticles as a promising drug delivery system, necessitating further investigations to explore their clinical applicability.
通过溶热法合成了纳米多孔铝金属有机框架(Al-MOF),并将其作为载体基质用于茵贝酮(Um)的体外给药。以 pH 值为 7.4 的模拟体液磷酸盐缓冲盐水(PBS)为释放介质,在 37 °C、7 天内逐渐释放被包裹的 Um。药物释放曲线表明,Al-MOF 纳米粒子具有作为有效药物递送载体的潜力。利用傅立叶变换红外光谱(FTIR)、X 射线衍射仪(XRD)和紫外可见光谱(UV-Vis)对负载 Um 的 Al-MOF 纳米粒子(Um-Al MOF)进行了结构和化学分析。热重分析(TGA)用于研究 Al-MOF 纳米粒子的热稳定性,而透射电子显微镜(TEM)则用于评估其形态特征。与单独的 Um 和 Al-MOF 纳米粒子相比,Um-Al MOF 纳米粒子具有显著的抗氧化和抗炎特性。此外,它们还在蚯蚓模型中显示出明显的伤口愈合能力。这些发现凸显了 Al-MOF 纳米粒子作为一种前景广阔的给药系统的潜力,因此有必要对其进行进一步研究,以探索其临床适用性。
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引用次数: 0
A marker-free genetic manipulation method for Glaesserella parasuis strains developed by alternately culturing transformants at 37°C and 30°C. 通过在 37°C 和 30°C 温度下交替培养转化株,开发出一种无标记的寄生褐藻菌株遗传操作方法。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-03 DOI: 10.1186/s12896-024-00887-w
Jing Xiao, Yuxin Wang, Dongfang Wu, Yuping Song, Xuwang Cai, Huanchun Chen, Hongbo Zhou, Xiaojuan Xu

Background: Glaesserella parasuis (G. parasuis) is the causative agent of Glässer's disease, which causes significant economic losses in the swine industry. However, research on the pathogenesis of G. parasuis has been hampered by the lack of a simple and efficient marker-free knockout system.

Results: In this study, a marker-free knockout system was developed for G. parasuis using a temperature-sensitive vector. By alternating the incubation of transformants at 30°C and 37°C, we optimized the screening process for this system. The system was successfully applied to knockout the KanR cassette from JS0135ΔnanH::KanR, achieving a knockout efficiency of 90% in the final round of screening. To confirm that temperature variation was a key factor, we proceeded with knocking out the nanH and apd genes in the CF7066 strain. The knockout efficiency reached up to 100%, with the shortest screening time being only four days. The knockout of the nanH gene resulted in a significant reduction in the growth vitality of the strains, while the knockout of the apd gene led to an approximate 56% improvement in the adhesion rate. Additionally, we observed that the expression of recombinant genes in transformants was higher at 30℃ than at 37℃, with the recC gene being upregulated approximately 7-fold. In contrast, there was almost no difference in the expression of recombinant genes between 30℃ and 37℃ in the wild-type strains. This discrepancy was likely due to an elevated copy number of target plasmids at 30℃, which may have resulted in the enhanced expression of recombinant genes.

Conclusions: In conclusion, this newly developed gene knockout system for G. parasuis presents a valuable tool for advancing research on this organism.

背景:寄生格氏菌(Glaesserella parasuis,G. parasuis)是格莱塞氏病的病原体,它给养猪业造成了巨大的经济损失。然而,由于缺乏简单高效的无标记基因敲除系统,对寄生璃泽氏菌发病机制的研究一直受到阻碍:结果:本研究利用对温度敏感的载体为寄生虫开发了一种无标记基因剔除系统。通过交替在 30°C 和 37°C 下培养转化子,我们优化了该系统的筛选过程。该系统被成功用于敲除 JS0135ΔnanH::KanR 中的 KanR 盒,在最后一轮筛选中,敲除效率达到了 90%。为了证实温度变化是一个关键因素,我们继续在 CF7066 菌株中敲除 nanH 和 apd 基因。基因敲除效率高达 100%,最短筛选时间仅为四天。敲除 nanH 基因会显著降低菌株的生长活力,而敲除 apd 基因则会使粘附率提高约 56%。此外,我们还观察到,重组基因在转化株中的表达量在 30℃ 比 37℃ 高,其中 recC 基因上调了约 7 倍。相比之下,野生型菌株重组基因的表达在 30℃ 和 37℃ 之间几乎没有差异。这种差异可能是由于 30℃ 时目的质粒的拷贝数增加,从而导致重组基因的表达增强:总之,这种新开发的寄生虫基因敲除系统是推进寄生虫研究的重要工具。
{"title":"A marker-free genetic manipulation method for Glaesserella parasuis strains developed by alternately culturing transformants at 37°C and 30°C.","authors":"Jing Xiao, Yuxin Wang, Dongfang Wu, Yuping Song, Xuwang Cai, Huanchun Chen, Hongbo Zhou, Xiaojuan Xu","doi":"10.1186/s12896-024-00887-w","DOIUrl":"10.1186/s12896-024-00887-w","url":null,"abstract":"<p><strong>Background: </strong>Glaesserella parasuis (G. parasuis) is the causative agent of Glässer's disease, which causes significant economic losses in the swine industry. However, research on the pathogenesis of G. parasuis has been hampered by the lack of a simple and efficient marker-free knockout system.</p><p><strong>Results: </strong>In this study, a marker-free knockout system was developed for G. parasuis using a temperature-sensitive vector. By alternating the incubation of transformants at 30°C and 37°C, we optimized the screening process for this system. The system was successfully applied to knockout the Kan<sup>R</sup> cassette from JS0135ΔnanH::Kan<sup>R</sup>, achieving a knockout efficiency of 90% in the final round of screening. To confirm that temperature variation was a key factor, we proceeded with knocking out the nanH and apd genes in the CF7066 strain. The knockout efficiency reached up to 100%, with the shortest screening time being only four days. The knockout of the nanH gene resulted in a significant reduction in the growth vitality of the strains, while the knockout of the apd gene led to an approximate 56% improvement in the adhesion rate. Additionally, we observed that the expression of recombinant genes in transformants was higher at 30℃ than at 37℃, with the recC gene being upregulated approximately 7-fold. In contrast, there was almost no difference in the expression of recombinant genes between 30℃ and 37℃ in the wild-type strains. This discrepancy was likely due to an elevated copy number of target plasmids at 30℃, which may have resulted in the enhanced expression of recombinant genes.</p><p><strong>Conclusions: </strong>In conclusion, this newly developed gene knockout system for G. parasuis presents a valuable tool for advancing research on this organism.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"60"},"PeriodicalIF":3.5,"publicationDate":"2024-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11373133/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142124721","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Purification and characterization of soluble recombinant Crimean-Congo hemorrhagic fever virus glycoprotein Gc expressed in mammalian 293F cells. 在哺乳动物 293F 细胞中表达的可溶性重组克里米亚-刚果出血热病毒糖蛋白 Gc 的纯化和表征。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-27 DOI: 10.1186/s12896-024-00885-y
Nigel Aminake Makoah, Matefo Millicent Litabe, Fredy Brice Nemg Simo, Katlego Keith Maboho, Felicity Jane Burt

Background: Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne zoonotic disease that presents with severe hemorrhagic manifestations and is associated with significant fatality rates. The causative agent, Crimean-Congo Hemorrhagic Fever Virus (CCHFV), is a high-priority pathogen identified by the World Health Organization with no approved vaccine or specific treatment available. In addition, there is a critical need for enhanced diagnostic tools to improve public health awareness, prevention measures, and disease control strategies.

Methods: We designed plasmids to enable the purification of soluble CCHFV glycoprotein Gc expressed in mammalian 293 F cells, followed by purification using affinity and size exclusion chromatography. The purified antigen was analyzed by SDS-PAGE and Western blotting to confirm its reactivity to antibodies from CCHF survivors. Additionally, an in-house indirect ELISA was developed using the purified Gc as a coating antigen.

Results: The optimized expression system successfully produced soluble and pure Gc antigen after affinity chromatography. The protein showed specific reactivity with CCHFV-positive serum antibodies in Western blot analysis. The indirect ELISA assay demonstrated high efficacy in distinguishing between CCHFV-positive and -negative serum samples, indicating its potential as a valuable diagnostic tool. Size exclusion chromatography further confirmed the presence of aggregates in our protein preparation.

Conclusions: The purified Gc antigen shows promise for developing direct diagnostic assays for CCHFV. The antigen's suitability for subunit vaccine development and its application as bait for monoclonal antibody isolation from survivors could be investigated further. This work lays the foundation for future research into the development of rapid diagnostic tests for field deployment.

背景:克里米亚-刚果出血热(CCHF)是一种由蜱虫传播的人畜共患病,表现为严重的出血性症状,致死率很高。病原体克里米亚-刚果出血热病毒(CCHFV)是世界卫生组织确定的高度优先病原体,目前还没有获得批准的疫苗或特效治疗方法。此外,亟需加强诊断工具,以提高公共卫生意识、改进预防措施和疾病控制策略:方法:我们设计了质粒来纯化在哺乳动物 293 F 细胞中表达的可溶性 CCHFV 糖蛋白 Gc,然后使用亲和层析和尺寸排阻层析进行纯化。纯化后的抗原通过 SDS-PAGE 和 Western 印迹法进行分析,以确认其与 CCHF 存活者的抗体的反应性。此外,还使用纯化的 Gc 作为包被抗原开发了一种内部间接 ELISA:结果:经过优化的表达系统在亲和层析后成功产生了可溶性纯Gc抗原。在 Western 印迹分析中,该蛋白与 CCHFV 阳性血清抗体呈特异性反应。间接酶联免疫吸附试验在区分 CCHFV 阳性和阴性血清样本方面表现出很高的效率,表明它有潜力成为一种有价值的诊断工具。尺寸排阻色谱法进一步证实了我们制备的蛋白质中存在聚集体:纯化的 Gc 抗原有望用于开发 CCHFV 的直接诊断测定。结论:纯化的 Gc 抗原有望用于开发 CCHFV 的直接诊断检测,该抗原是否适合亚单位疫苗的开发,以及是否可用作从存活者中分离单克隆抗体的诱饵,还有待于进一步研究。这项工作为今后研究开发用于野外部署的快速诊断测试奠定了基础。
{"title":"Purification and characterization of soluble recombinant Crimean-Congo hemorrhagic fever virus glycoprotein Gc expressed in mammalian 293F cells.","authors":"Nigel Aminake Makoah, Matefo Millicent Litabe, Fredy Brice Nemg Simo, Katlego Keith Maboho, Felicity Jane Burt","doi":"10.1186/s12896-024-00885-y","DOIUrl":"10.1186/s12896-024-00885-y","url":null,"abstract":"<p><strong>Background: </strong>Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne zoonotic disease that presents with severe hemorrhagic manifestations and is associated with significant fatality rates. The causative agent, Crimean-Congo Hemorrhagic Fever Virus (CCHFV), is a high-priority pathogen identified by the World Health Organization with no approved vaccine or specific treatment available. In addition, there is a critical need for enhanced diagnostic tools to improve public health awareness, prevention measures, and disease control strategies.</p><p><strong>Methods: </strong>We designed plasmids to enable the purification of soluble CCHFV glycoprotein Gc expressed in mammalian 293 F cells, followed by purification using affinity and size exclusion chromatography. The purified antigen was analyzed by SDS-PAGE and Western blotting to confirm its reactivity to antibodies from CCHF survivors. Additionally, an in-house indirect ELISA was developed using the purified Gc as a coating antigen.</p><p><strong>Results: </strong>The optimized expression system successfully produced soluble and pure Gc antigen after affinity chromatography. The protein showed specific reactivity with CCHFV-positive serum antibodies in Western blot analysis. The indirect ELISA assay demonstrated high efficacy in distinguishing between CCHFV-positive and -negative serum samples, indicating its potential as a valuable diagnostic tool. Size exclusion chromatography further confirmed the presence of aggregates in our protein preparation.</p><p><strong>Conclusions: </strong>The purified Gc antigen shows promise for developing direct diagnostic assays for CCHFV. The antigen's suitability for subunit vaccine development and its application as bait for monoclonal antibody isolation from survivors could be investigated further. This work lays the foundation for future research into the development of rapid diagnostic tests for field deployment.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"59"},"PeriodicalIF":3.5,"publicationDate":"2024-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11348531/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142078967","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Correction: Comparison of lipidome profiles in serum from lactating dairy cows supplemented with Acremonium terrestris culture based on UPLC-QTRAP-MS/MS. 更正:基于 UPLC-QTRAP-MS/MS 的泌乳奶牛血清脂质体图谱比较。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-22 DOI: 10.1186/s12896-024-00886-x
Chenmiao Zhang, Yiran Zhao, Shijiao Guo, Feifei Li, Xu Gong, Jiarui Gao, Linshu Jiang, Jinjin Tong
{"title":"Correction: Comparison of lipidome profiles in serum from lactating dairy cows supplemented with Acremonium terrestris culture based on UPLC-QTRAP-MS/MS.","authors":"Chenmiao Zhang, Yiran Zhao, Shijiao Guo, Feifei Li, Xu Gong, Jiarui Gao, Linshu Jiang, Jinjin Tong","doi":"10.1186/s12896-024-00886-x","DOIUrl":"10.1186/s12896-024-00886-x","url":null,"abstract":"","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"57"},"PeriodicalIF":3.5,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11361175/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142091842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Nitric oxide mediates positive regulation of Nostoc flagelliforme polysaccharide yield via potential S-nitrosylation of G6PDH and UGDH. 一氧化氮通过对 G6PDH 和 UGDH 潜在的 S-亚硝基化作用,介导对 Nostoc flagelliforme 多糖产量的正向调节。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-22 DOI: 10.1186/s12896-024-00884-z
Meng-Yuan Li, Yan-Ru Li, Cheng-Feng Han, Jie Zhang, Rui-Ying Zhu, Yan Zhang, Jian Li, Shi-Ru Jia, Pei-Pei Han

Based on our previous findings that salicylic acid and jasmonic acid increased Nostoc flagelliforme polysaccharide yield by regulating intracellular nitric oxide (NO) levels, the mechanism through which NO affects polysaccharide biosynthesis in Nostoc flagelliforme was explored from the perspective of S-nitrosylation (SNO). The addition of NO donor and scavenger showed that intracellular NO had a significant positive effect on the polysaccharide yield of N. flagelliforme. To explore the mechanism, we investigated the relationship between NO levels and the activity of several key enzymes involved in polysaccharide biosynthesis, including fructose 1,6-bisphosphate aldolase (FBA), glucokinase (GK), glucose 6-phosphate dehydrogenase (G6PDH), mitochondrial isocitrate dehydrogenase (ICDH), and UDP-glucose dehydrogenase (UGDH). The enzymatic activities of G6PDH, ICDH, and UGDH were shown to be significantly correlated with the shifts in intracellular NO levels. For further validation, G6PDH, ICDH, and UGDH were heterologously expressed in Escherichia coli and purified via Ni+-NAT affinity chromatography, and subjected to a biotin switch assay and western blot analysis, which revealed that UGDH and G6PDH were susceptible to SNO. Furthermore, mass spectrometry analysis of proteins treated with S-nitrosoglutathione (GSNO) identified the SNO modification sites for UGDH and G6PDH as cysteine 423 and cysteine 249, respectively. These findings suggest that NO modulates polysaccharide biosynthesis in N. flagelliforme through SNO of UGDH and G6PDH. This reveals a potential mechanism through which NO promotes polysaccharide synthesis in N. flagelliforme, while also providing a new strategy for improving the industrial production of polysaccharides.

基于水杨酸和茉莉酸通过调节细胞内一氧化氮(NO)水平增加鞭毛藻多糖产量的研究结果,我们从S-亚硝基化(SNO)的角度探讨了NO影响鞭毛藻多糖生物合成的机制。加入 NO 供体和清除剂后发现,细胞内 NO 对鞭毛藻多糖产量有显著的正向影响。为了探索其机制,我们研究了 NO 水平与参与多糖生物合成的几种关键酶活性之间的关系,包括 1,6-二磷酸果糖醛缩酶 (FBA)、葡萄糖激酶 (GK)、6-磷酸葡萄糖脱氢酶 (G6PDH)、线粒体异柠檬酸脱氢酶 (ICDH) 和 UDP-葡萄糖脱氢酶 (UGDH)。研究表明,G6PDH、ICDH 和 UGDH 的酶活性与细胞内 NO 水平的变化显著相关。为了进一步验证,在大肠杆菌中异源表达了 G6PDH、ICDH 和 UGDH,并通过 Ni+-NAT 亲和层析进行纯化,然后进行生物素转换测定和 Western 印迹分析,结果显示 UGDH 和 G6PDH 易受 SNO 影响。此外,经 S-亚硝基谷胱甘肽(GSNO)处理的蛋白质的质谱分析表明,UGDH 和 G6PDH 的 SNO 修饰位点分别为半胱氨酸 423 和半胱氨酸 249。这些发现表明,氮氧化物通过对 UGDH 和 G6PDH 的 SNO 来调节鞭毛虫多糖的生物合成。这揭示了 NO 促进鞭毛菜多糖合成的潜在机制,同时也为改善多糖的工业生产提供了新策略。
{"title":"Nitric oxide mediates positive regulation of Nostoc flagelliforme polysaccharide yield via potential S-nitrosylation of G6PDH and UGDH.","authors":"Meng-Yuan Li, Yan-Ru Li, Cheng-Feng Han, Jie Zhang, Rui-Ying Zhu, Yan Zhang, Jian Li, Shi-Ru Jia, Pei-Pei Han","doi":"10.1186/s12896-024-00884-z","DOIUrl":"10.1186/s12896-024-00884-z","url":null,"abstract":"<p><p>Based on our previous findings that salicylic acid and jasmonic acid increased Nostoc flagelliforme polysaccharide yield by regulating intracellular nitric oxide (NO) levels, the mechanism through which NO affects polysaccharide biosynthesis in Nostoc flagelliforme was explored from the perspective of S-nitrosylation (SNO). The addition of NO donor and scavenger showed that intracellular NO had a significant positive effect on the polysaccharide yield of N. flagelliforme. To explore the mechanism, we investigated the relationship between NO levels and the activity of several key enzymes involved in polysaccharide biosynthesis, including fructose 1,6-bisphosphate aldolase (FBA), glucokinase (GK), glucose 6-phosphate dehydrogenase (G6PDH), mitochondrial isocitrate dehydrogenase (ICDH), and UDP-glucose dehydrogenase (UGDH). The enzymatic activities of G6PDH, ICDH, and UGDH were shown to be significantly correlated with the shifts in intracellular NO levels. For further validation, G6PDH, ICDH, and UGDH were heterologously expressed in Escherichia coli and purified via Ni<sup>+</sup>-NAT affinity chromatography, and subjected to a biotin switch assay and western blot analysis, which revealed that UGDH and G6PDH were susceptible to SNO. Furthermore, mass spectrometry analysis of proteins treated with S-nitrosoglutathione (GSNO) identified the SNO modification sites for UGDH and G6PDH as cysteine 423 and cysteine 249, respectively. These findings suggest that NO modulates polysaccharide biosynthesis in N. flagelliforme through SNO of UGDH and G6PDH. This reveals a potential mechanism through which NO promotes polysaccharide synthesis in N. flagelliforme, while also providing a new strategy for improving the industrial production of polysaccharides.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"58"},"PeriodicalIF":3.5,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11342573/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142035140","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Recombinant hirudin and PAR-1 regulate macrophage polarisation status in diffuse large B-cell lymphoma 重组水蛭素和 PAR-1 可调节弥漫大 B 细胞淋巴瘤中巨噬细胞的极化状态
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-12 DOI: 10.1186/s12896-024-00879-w
Qiang Pei, Zihui Li, Jingjing Zhao, Haixi Zhang, Tao Qin, Juan Zhao
Diffuse large B-cell lymphoma (DLBCL) is a malignant tumour. Although some standard therapies have been established to improve the cure rate, they remain ineffective for specific individuals. Therefore, it is meaningful to find more novel therapeutic approaches. Macrophage polarisation is extensively involved in the process of tumour development. Recombinant hirudin (rH) affects macrophages and has been researched frequently in clinical trials lately. Our article validated the regulatory role of rH in macrophage polarisation and the mechanism of PAR-1 by collecting clinical samples and subsequently establishing a cellular model to provide a scientifically supported perspective for discovering new therapeutic approaches. We assessed the expression of macrophage polarisation markers, cytokines and PAR-1 in clinical samples. We established a cell model by co-culture with THP-1 and OCI-Ly10 cell. We determined the degree of cell polarisation and expression of validation cytokines by flow cytometry, ELISA, and RT-qPCR to confirm the success of the cell model. Subsequently, different doses of rH were added to discover the function of rH on cell polarisation. We confirmed the mechanism of PAR-1 in macrophage polarisation by transfecting si-PAR-1 and pcDNA3.1-PAR-1. We found higher expression of M2 macrophage markers (CD163 + CMAF+) and PAR-1 in 32 DLBCL samples. After inducing monocyte differentiation into M0 macrophages and co-culturing with OCI-Ly10 lymphoma cells, we found a trend of these expressions in the cell model consistent with the clinical samples. Subsequently, we discovered that rH promotes the polarisation of M1 macrophages but inhibits the polarisation of M2 macrophages. We also found that PAR-1 regulates macrophage polarisation, inhibiting cell proliferation, migration, invasion and angiogenic capacity. rH inhibits macrophage polarisation towards the M2 type and PAR-1 regulates polarisation, proliferation, migration, invasion, and angiogenesis of DLBCL-associated macrophages.
弥漫大 B 细胞淋巴瘤(DLBCL)是一种恶性肿瘤。虽然一些标准疗法已被确立以提高治愈率,但它们对特定个体仍然无效。因此,寻找更多新的治疗方法意义重大。巨噬细胞极化广泛参与了肿瘤的发展过程。重组水蛭素(rH)会影响巨噬细胞,最近在临床试验中被频繁研究。我们的文章通过收集临床样本验证了 rH 在巨噬细胞极化中的调控作用以及 PAR-1 的作用机制,并随后建立了一个细胞模型,为发现新的治疗方法提供了一个有科学依据的视角。我们评估了临床样本中巨噬细胞极化标志物、细胞因子和 PAR-1 的表达。我们通过与 THP-1 和 OCI-Ly10 细胞共培养建立了细胞模型。我们通过流式细胞术、ELISA 和 RT-qPCR 测定了细胞极化的程度和验证细胞因子的表达,以确认细胞模型的成功。随后,我们加入了不同剂量的 rH,以发现 rH 对细胞极化的作用。我们通过转染 si-PAR-1 和 pcDNA3.1-PAR-1 证实了 PAR-1 在巨噬细胞极化中的作用机制。我们发现在32个DLBCL样本中,M2巨噬细胞标志物(CD163 + CMAF+)和PAR-1的表达较高。在诱导单核细胞分化为 M0 巨噬细胞并与 OCI-Ly10 淋巴瘤细胞共培养后,我们发现细胞模型中这些表达的趋势与临床样本一致。随后,我们发现 rH 可促进 M1 巨噬细胞的极化,但会抑制 M2 巨噬细胞的极化。我们还发现,PAR-1 可调节巨噬细胞的极化,抑制细胞的增殖、迁移、侵袭和血管生成能力。
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引用次数: 0
Comparison of lipidome profiles in serum from lactating dairy cows supplemented with Acremonium terrestris culture based on UPLC-QTRAP-MS/MS 基于 UPLC-QTRAP-MS/MS 的泌乳奶牛血清脂质体谱分析比较
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-12 DOI: 10.1186/s12896-024-00881-2
Chenmiao Zhang, Yiran Zhao, Shijiao Guo, Feifei Li, Xu Gong, Jiarui Gao, Linshu Jiang, Jinjin Tong
This study evaluated the effects of supplementing the diet of lactating cows with Acremonium terrestris culture (ATC) on milk production, serum antioxidant capacity, inflammatory indices, and serum lipid metabolomics. Over 90 days, 24 multiparous Chinese Holstein cows in mid-lactation (108 ± 10.4 days in milk, 637 ± 25 kg body weight, 30.23 ± 3.7 kg/d milk yield) were divided into either a control diet (CON) or a diet supplemented with 30 g of ATC daily. All the data were analyzed using Student’s t test with SPSS 20.0 software. The results showed that compared with CON feeding, ATC feeding significantly increased milk yield, antioxidant capacity, and immune function. Lipidome screening identified 143 lipid metabolites that differed between the two groups. Further analysis using “random forest” machine learning revealed three glycerophospholipid serum metabolites that could serve as lipid markers with a predictive accuracy of 91.67%. This study suggests that ATC can be a useful dietary supplement for improving lactational performance in dairy cows and provides valuable insights into developing nutritional strategies to maintain metabolic homeostasis in ruminants.
本研究评估了在泌乳奶牛的日粮中添加白藜芦醇培养物(ATC)对产奶量、血清抗氧化能力、炎症指数和血清脂质代谢组学的影响。在 90 天内,将 24 头处于泌乳中期的多胎中国荷斯坦奶牛(108 ± 10.4 天,637 ± 25 千克体重,30.23 ± 3.7 千克/天产奶量)分为对照日粮(CON)或每天添加 30 克 ATC 的日粮。所有数据均采用 SPSS 20.0 软件的学生 t 检验进行分析。结果表明,与对照组相比,ATC能显著提高产奶量、抗氧化能力和免疫功能。脂质体筛选确定了两组之间存在差异的 143 种脂质代谢物。利用 "随机森林 "机器学习进行的进一步分析发现,有三种甘油磷脂血清代谢物可作为脂质标记物,预测准确率高达 91.67%。这项研究表明,ATC 可以作为一种有用的膳食补充剂来提高奶牛的泌乳性能,并为制定维持反刍动物代谢平衡的营养策略提供了有价值的见解。
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BMC Biotechnology
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