Biotechnology Notes最新文献_第3页

Agroinfiltration-mediated transient assay for rapid evaluation of constructs in pigeonpea 农业浸润介导的快速评价鸽豌豆构建体的瞬时试验

Biotechnology Notes

Pub Date : 2025-01-01 DOI: 10.1016/j.biotno.2025.02.005

Kalenahalli Yogendra, Harika Gadeela, Koppula Nithya Sree, Wricha Tyagi

The process of generating stable transformants is time-consuming, labor-intensive, and genotype-dependent. In contrast, transient gene expression techniques, such as agroinfiltration, offer a rapid assessment of gene function and expression. Agroinfiltration, widely employed for studying gene function, has been extensively applied in leaf tissues of Nicotiana benthamiana and various other plant species. Despite its broad utility in various plants, to our knowledge, no prior investigation has been reported in pigeonpea. In this study, we developed an agroinfiltration method for transiently expressing a green fluorescent protein (mGFP5) reporter gene in four pigeonpea genotypes using syringe infiltration at the seedling stage under greenhouse conditions. The expression of the reporter gene mGFP5 was assessed at 72-, 96-, and 120 h post-infiltration (hpi). Additionally, we assessed the effect of morphogenic genes, specifically growth-regulating factor 4 (GRF4) and GRF-interacting factor 1 (GIF1), from both rice and pigeonpea on the expression of mGFP5 in four pigeonpea genotypes. Our findings demonstrate that OsGRF4-GIF1 led to enhanced mGFP5 expression compared to CcGRF4-GIF1 in four diverse pigeonpea genotypes. Fluorescence could be detected till 120 hpi. Furthermore, PCR, RT-PCR, and fluorescence quantification confirmed the presence and expression of mGFP5 at 72 hpi. Our results highlight the efficacy of agroinfiltration in quickly evaluating candidate genes in four genetically diverse pigeonpea genotypes, thereby reducing the time required for the initial assessment of constructs suitable for diverse molecular biology analyses.

产生稳定的转化体的过程是耗时的，劳动密集型的，并且依赖于基因型。相比之下，瞬时基因表达技术，如农业渗透，提供了基因功能和表达的快速评估。农业渗透技术被广泛应用于研究基因功能，已广泛应用于烟叶和其他植物的叶组织中。尽管它在各种植物中广泛应用，但据我们所知，尚无关于鸽子豆的研究报道。在这项研究中，我们建立了一种农业渗透方法，在温室条件下，在苗期使用注射器渗透，在四种基因型的鸽子豌豆中瞬时表达绿色荧光蛋白（mGFP5）报告基因。在浸润后72、96和120 h （hpi）评估报告基因mGFP5的表达。此外，我们评估了水稻和鸽豌豆中形态发生基因，特别是生长调节因子4 （GRF4）和grf相互作用因子1 （GIF1）对四种鸽豌豆基因型中mGFP5表达的影响。我们的研究结果表明，与CcGRF4-GIF1相比，OsGRF4-GIF1在四种不同的鸽豆基因型中导致mGFP5的表达增强。直到120 hpi都能检测到荧光。此外，PCR、RT-PCR和荧光定量证实了72hpi时mGFP5的存在和表达。我们的研究结果强调了农业渗透在四种遗传多样性的鸽豌豆基因型中快速评估候选基因的有效性，从而减少了初始评估适合不同分子生物学分析的构建体所需的时间。

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引用次数: 0

Straightforward MALDI-TOF MS based screening approach for selection of recombinant protein-expressing E. coli 直接基于MALDI-TOF MS筛选表达重组蛋白大肠杆菌的方法

Biotechnology Notes

Pub Date : 2025-01-01 DOI: 10.1016/j.biotno.2025.02.004

I.N. Kravtsov , A.I. Solovyev , E.A. Potemkina , A.V. Kartashova , M.A. Dmitrieva , K.V. Danilova , I.L. Tutykhina , N.B. Polyakov , V.D. Desinov , D.A. Egorova , A.L. Gintsburg

Recombinant protein production is a milestone of modern biotechnology, drug development and scientific research. When obtaining recombinant protein producers, differences in expression levels among clones necessitate screening. Traditional widely used methods include protein electrophoresis and western blot hybridization. This protocol provides high-throughput advantages by eliminating time-consuming steps inherent to traditional methods, such as cell lysis, protein extraction, purification, antibody-based detection, and gel-based analysis. MALDI-TOF MS represents a simple, rapid and cost-effective method for bacterial species identification through protein fingerprint signature in clinical diagnostics, but not practically integrated into biotechnological workflow. This study proposes a fast and easy method for screening E. coli clones producing recombinant proteins with MALDI-TOF MS. The proposed method demonstrated efficiency in screening of E. coli producing several recombinant proteins with different properties: sfGFP; bacterial DNA binding proteins IHFα, IHFβ, HU; bacteriophage protein GP46 and camelid VHH antibody fragments.

重组蛋白的生产是现代生物技术、药物开发和科学研究的一个里程碑。在获得重组蛋白生产者时，需要筛选克隆之间表达水平的差异。传统的广泛使用的方法包括蛋白质电泳和western blot杂交。该方案通过消除传统方法固有的耗时步骤提供高通量优势，例如细胞裂解，蛋白质提取，纯化，基于抗体的检测和基于凝胶的分析。MALDI-TOF质谱在临床诊断中是一种简单、快速、具有成本效益的蛋白质指纹图谱细菌种类鉴定方法，但尚未真正融入生物技术工作流程。本研究提出了一种快速简便的利用MALDI-TOF ms筛选产生重组蛋白的大肠杆菌克隆的方法，该方法在筛选产生不同性质重组蛋白的大肠杆菌中表现出高效率：sfGFP；细菌DNA结合蛋白IHFα、IHFβ、HU；噬菌体蛋白GP46和骆驼类VHH抗体片段。

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引用次数: 0

Tackling carbapenem-resistant Acinetobacter baumannii (CRAB) and their virulence factors using biosynthesized silver nanoparticles combined with imipenem 生物合成纳米银与亚胺培南联合处理耐碳青霉烯鲍曼不动杆菌及其毒力因子

Biotechnology Notes

Pub Date : 2025-01-01 DOI: 10.1016/j.biotno.2025.07.002

Mohamed Shawky, Mohamed H. Kalaba, Gamal M. El-Sherbiny

Carbapenem-resistant Acinetobacter baumannii is an extremely hazardous microorganism due to its high level of resistance to a wide array of antibiotics, making it a significant public health concern. Herein, this study aimed to biofabricate silver nanoparticles using a cell-free filter derived from Streptomyces sp., with a focus on characterizing their physicochemical properties, and use them to combat CRAB and their virulence factors. The biofabricated Ag-NPs were predominantly spherical with an average size 50 nm, confirmed through TEM analyses, while DLS measurements showed an average hydrodynamic diameter of approximately 36.78 nm. UV–Vis spectroscopy displayed a characteristic surface plasmon resonance peak in the range of 420 nm, indicative of nanoparticle formation. XRD confirmed the crystalline structure, presenting peaks corresponding to face-centered cubic silver. FTIR spectroscopy revealed active participation of metabolite compounds derived from the Streptomyces cell-free filter in both reduction and stabilization processes. Eight clinical bacterial isolates were identified as CRAB using the Vitek-2 system, and biofilm formation with 100 % was assessed through Congo red and microplate assays. The MIC for Ag-NPs and imipenem (IMP) were found to be between 4 and 5 μg/mL and 13 and 15 μg/mL, respectively. Additionally, the fractional inhibitory concentration index (FICI) for the synergistic combinations of Ag-NPs and IMP ranged from 0.5 to 0.375, indicating a notable decrease in the MIC values for both IMP and Ag-NPs from 14 and 5 μg/mL to 1.75 and 1.25 μg/mL, respectively. The qRT-PCR demonstrated a significant reduction in the expression levels of the Bap and ompA genes by up to 4.0-fold (p ≤ 0.001). The time-killing assay confirmed that the bacterial strain was effectively eliminated through the synergistic action of Ag-NPs and IMP. Moreover, the cytotoxicity assessment of Ag-NPs and their combination with IMP revealed low toxicity of the combination of Ag-NPs and IMP, with an IC₅₀ of 26.13 ± 0.24 and 45.33 ± 0.21 μg/mL, respectively (p < 0.0019), indicating good biosafety, while the hemolysis rates were recorded at 0.4 and 0.7 at 12 and 24 h, respectively. We concluded that the combination of Ag-NPs with IMP could serve as a promising alternative strategy for treating CRAB.

耐碳青霉烯鲍曼不动杆菌是一种极其危险的微生物，因为它对多种抗生素具有高度耐药性，使其成为一个重大的公共卫生问题。本研究旨在利用链霉菌（Streptomyces sp.）衍生的无细胞过滤器生物制备纳米银，重点研究其物理化学性质，并利用其对抗螃蟹及其毒力因子。通过TEM分析证实，生物制备的Ag-NPs主要为球形，平均尺寸为50 nm，而DLS测量显示其平均水动力直径约为36.78 nm。紫外可见光谱显示，表面等离子体共振峰在420 nm范围内，表明纳米颗粒形成。XRD证实其晶体结构为面心立方银。FTIR光谱显示，链霉菌无细胞过滤器衍生的代谢物化合物积极参与还原和稳定过程。使用Vitek-2系统鉴定8株临床分离细菌为CRAB，通过刚果红和微孔板检测100%的生物膜形成情况。Ag-NPs和亚胺培南（IMP）的MIC分别为4 ~ 5 μg/mL和13 ~ 15 μg/mL。此外，Ag-NPs和IMP协同组合的分数抑制浓度指数（FICI）在0.5 ~ 0.375之间，表明IMP和Ag-NPs的MIC值分别从14和5 μg/mL显著降低到1.75和1.25 μg/mL。qRT-PCR结果显示，Bap和ompA基因的表达水平显著降低了4.0倍（p≤0.001）。时间杀伤实验证实，Ag-NPs与IMP的协同作用可有效杀灭该菌株，细胞毒性评价显示Ag-NPs与IMP联用毒性较低，IC50分别为26.13±0.24和45.33±0.21 μg/mL (p <；0.0019)，生物安全性较好，12 h和24 h溶血率分别为0.4和0.7。我们的结论是Ag-NPs与IMP的联合治疗可能是一种有希望的治疗螃蟹的替代策略。

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引用次数: 0

Modeling and simulation of soft bio-mimetic fingers with a novel soft thumb design for bionic hand applications using ANN 基于人工神经网络的柔性仿生手指建模与仿真研究

Biotechnology Notes

Pub Date : 2025-01-01 DOI: 10.1016/j.biotno.2025.05.002

Eman R.A. Soliman , Ayman Nada , Hiroyuki Ishii , Ahmed M.R. Fathelbeb

The paper presents a novel design for a soft bio-mimetic finger and soft thumb structure for bionic hand applications. It introduces an anthropomorphic pneumatic flexible finger system using a PneuNets framework to enhance flexibility and maneuverability. The research investigates the influence of geometric variations (wall thickness, chamber number, and spacing) on finger deformation, demonstrating that reduced wall thickness and augmented chambers substantially improve flexibility. A key innovation is the soft thumb design that accurately replicates the complex movements of the Carpometacarpal (CMC) joint, enabling natural opposition and dexterity. Eight models were developed for four fingers and two models for the thumb. Simulation results indicate that models with thinner walls (2 mm) achieve bending angles exceeding 80° at 120 KPa, whereas 3 mm models remain below 50°. Moreover, increasing the number of chambers enhances deformation, with each added chamber contributing approximately 41 % more flexibility. For the thumb models, we successfully mapped the motion ranges and accurately mimicked the base joint, enabling natural opposition and dexterity. Furthermore, the paper also integrates Artificial Neural Networks (ANNs) to model forward kinematics, improving the estimation of bending angles and end-tip positions, which enhances the overall adaptability and control of the system.

本文提出了一种适用于仿生手的柔性仿生手指和柔性拇指结构设计。采用PneuNets框架设计了一种拟人气动柔性手指系统，提高了系统的灵活性和可操作性。该研究调查了几何变化（壁厚、室数和间距）对手指变形的影响，表明减少壁厚和增加室可大大提高灵活性。一个关键的创新是柔软的拇指设计，准确地复制了复杂的腕掌骨（CMC）关节的运动，使自然对立和灵巧。为四个手指开发了八个模型，为拇指开发了两个模型。仿真结果表明，在120 KPa下，壁较薄（2 mm）的模型的弯曲角超过80°，而3 mm的模型的弯曲角低于50°。此外，增加腔室的数量可以增强变形，每增加一个腔室，可使弹性提高约41%。对于拇指模型，我们成功地绘制了运动范围，并准确地模拟了基础关节，实现了自然的对立和灵巧性。此外，本文还将人工神经网络（ann）集成到机器人的正运动学建模中，改进了机器人弯曲角度和端尖位置的估计，增强了系统的整体适应性和控制能力。

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引用次数: 0

Cell-free protein synthesis platforms for accelerating drug discovery 加速药物发现的无细胞蛋白质合成平台

Biotechnology Notes

Pub Date : 2025-01-01 DOI: 10.1016/j.biotno.2025.02.001

Filippo Caschera

Cell-free protein synthesis is a platform for streamlined production of macromolecules. Recently, several proteins with pharmaceutical relevance were synthesised and characterised. Off-the-shelf reagents and parallelised experimentation have enabled the exploration of many different conditions for in vitro protein synthesis and engineering. Herein is described how machine learning algorithms were applied for protein yield maximisation as well as for protein engineering and de novo design. Cell-free protein synthesis provides the biotechnological platform to unlock the power and benefit of AI/ML for drug discovery and improve human health.

无细胞蛋白质合成是大分子流线型生产的平台。最近，几种与药物相关的蛋白质被合成和表征。现成的试剂和并行实验使得探索体外蛋白质合成和工程的许多不同条件成为可能。本文描述了机器学习算法如何应用于蛋白质产量最大化以及蛋白质工程和从头设计。无细胞蛋白合成提供了生物技术平台，以释放AI/ML的力量和益处，用于药物发现和改善人类健康。

引用次数: 0

Mevalonate secretion is not mediated by a singular non-essential transporter in Saccharomyces cerevisiae 麦角酵母中的甲羟戊酸分泌不是由单一的非必要转运体介导的

Biotechnology Notes

Pub Date : 2024-01-01 DOI: 10.1016/j.biotno.2024.10.001

Scott A. Wegner , José L. Avalos

Isoprenoids are highly valued targets for microbial chemical production, allowing the creation of fragrances, biofuels, and pharmaceuticals from renewable carbon feedstocks. To increase isoprenoid production, previous efforts have manipulated pyruvate dehydrogenase (PDH) bypass pathway flux to increase cytosolic acetyl-coA; however, this results in mevalonate secretion and does not necessarily translate into higher isoprenoid production. Identification and disruption of the transporter mediating mevalonate secretion would allow us to determine whether increasing PDH bypass activity in the absence of secretion improves conversion of mevalonate into downstream isoprenoids. Attempted identification of the mevalonate transporter was accomplished using a pooled CRISPR library targeting all nonessential transporters and two different screening methods. Using a high throughput screen, based on growth of a mevalonate auxotrophic Escherichia coli strain, it was found that ZRT3 disruption largely abolished accumulation of extracellular mevalonate. However, disruption of ZRT3 was found to lower overall mevalonate pathway activity, rather than prevent secretion, indicating a previously unreported interaction between zinc availability and the mevalonate pathway. In a second screen, significant differences in PDR5/15 and QDR1/2 library representation were found between wild-type and mevalonate secreting Saccharomyces cerevisiae strains. However, no single deletion (or selected pair of double deletions) abolishes mevalonate secretion, indicating that this process appears to be mediated through multiple redundant transporters.

异戊烯类化合物是微生物化学生产的重要目标，可利用可再生碳原料生产香料、生物燃料和药物。为了提高异戊烯类化合物的产量，以前的研究曾通过操纵丙酮酸脱氢酶（PDH）旁路通量来增加细胞膜乙酰-coA；但这会导致甲羟戊酸的分泌，并不一定会转化为更高的异戊烯类化合物产量。鉴定和破坏介导甲羟戊酸分泌的转运体将使我们能够确定，在没有甲羟戊酸分泌的情况下，增加 PDH 旁路活性是否会改善甲羟戊酸向下游异丙烯酸的转化。我们尝试使用针对所有非必要转运体的 CRISPR 文库和两种不同的筛选方法来鉴定甲羟戊酸转运体。通过高通量筛选，在甲羟戊酸辅助营养大肠杆菌菌株生长的基础上发现，ZRT3 的破坏在很大程度上抑制了细胞外甲羟戊酸的积累。然而，研究发现破坏 ZRT3 会降低甲羟戊酸通路的整体活性，而不是阻止分泌，这表明锌的可用性与甲羟戊酸通路之间存在一种以前未报道过的相互作用。在第二个筛选中，发现野生型和分泌甲羟戊酸的酿酒酵母菌株之间的 PDR5/15 和 QDR1/2 文库代表性存在显著差异。然而，没有任何单个缺失（或选定的一对双缺失）会取消甲羟戊酸的分泌，这表明这一过程似乎是通过多个冗余转运体介导的。

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引用次数: 0

The potential of copper oxide nanoparticles in nanomedicine: A comprehensive review 氧化铜纳米粒子在纳米医学中的潜力：综述

Biotechnology Notes

Pub Date : 2024-01-01 DOI: 10.1016/j.biotno.2024.06.001

Mahalakshmi Devaraji, Punniyakoti V. Thanikachalam, Karthikeyan Elumalai

Nanotechnology is a modern scientific discipline that uses nanoparticles of metals like copper, silver, gold, platinum, and zinc for various applications. Copper oxide nanoparticles (CuONPs) are effective in biomedical settings, such as killing bacteria, speeding up reactions, stopping cancer cells, and coating surfaces. These inorganic nanostructures have a longer shelf life than their organic counterparts and are chemically inert and thermally stable. However, commercial synthesis of NPs often involves harmful byproducts and hazardous chemicals. Green synthesis for CuONPs offers numerous benefits, including being clean, harmless, economical, and environmentally friendly. Using naturally occurring organisms like bacteria, yeast, fungi, algae, and plants can make CuONPs more environmentally friendly. CuONPs are expected to be used in nanomedicine due to their potent antimicrobial properties and disinfecting agents for infectious diseases. This comprehensive review looks to evaluate research articles published in the last ten years that investigate the antioxidant, anticancer, antibacterial, wound healing, dental application and catalytic properties of copper nanoparticles generated using biological processes. Utilising the scientific approach of large-scale data analytics. However, their toxic effects on vertebrates and invertebrates raise concerns about their use for diagnostic and therapeutic purposes. Therefore, biocompatibility and non-toxicity are crucial for selecting nanoparticles for clinical research.

纳米技术是一门将铜、银、金、铂和锌等金属纳米粒子用于各种应用的现代科学学科。氧化铜纳米粒子（CuONPs）在生物医学领域非常有效，如杀灭细菌、加速反应、阻止癌细胞和表面涂层。这些无机纳米结构比有机纳米结构的保质期更长，而且具有化学惰性和热稳定性。然而，NPs 的商业合成通常涉及有害副产品和危险化学品。CuONPs 的绿色合成具有清洁、无害、经济和环保等诸多优点。利用细菌、酵母、真菌、藻类和植物等天然生物可使 CuONPs 更加环保。由于 CuONPs 具有强大的抗菌特性，可作为传染病的消毒剂，因此有望用于纳米医学。本综述旨在评估过去十年中发表的研究文章，这些文章研究了利用生物过程生成的纳米铜粒子的抗氧化、抗癌、抗菌、伤口愈合、牙科应用和催化特性。利用大规模数据分析的科学方法。然而，纳米铜粒子对脊椎动物和无脊椎动物的毒性作用引起了人们对其诊断和治疗用途的担忧。因此，生物相容性和无毒性是选择用于临床研究的纳米粒子的关键。

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引用次数: 0

aXonica: A support package for MRI based Neuroimaging aXonica：基于核磁共振成像的神经成像支持软件包

Biotechnology Notes

Pub Date : 2024-01-01 DOI: 10.1016/j.biotno.2024.08.001

Bilal Wajid , Momina Jamil , Fahim Gohar Awan , Faria Anwar , Ali Anwar

Magnetic Resonance Imaging (MRI) assists in studying the nervous system. MRI scans undergo significant processing before presenting the final images to medical practitioners. These processes are executed with ease due to excellent software pipelines. However, establishing software workstations is non-trivial and requires researchers in life sciences to be comfortable in downloading, installing, and scripting software that is non-user-friendly and may lack basic GUI. As researchers struggle with these skills, there is a dire need to develop software packages that can automatically install software pipelines speeding up building software workstations and laboratories. Previous solutions include NeuroDebian, BIDS Apps, Flywheel, QMENTA, Boutiques, Brainlife and Neurodesk. Overall, all these solutions complement each other. NeuroDebian covers neuroscience and has a wider scope, providing only 51 tools for MRI. Whereas, BIDS Apps is committed to the BIDS format, covering only 45 software related to MRI. Boutiques is more flexible, facilitating its pipelines to be easily installed as separate containers, validated, published, and executed. Whereas, both Flywheel and Qmenta are propriety, leaving four for users looking for ‘free for use’ tools, i.e., NeuroDebian, Brainlife, Neurodesk, and BIDS Apps. This paper presents an extensive survey of 317 tools published in MRI-based neuroimaging in the last ten years, along with ‘aXonica,’ an MRI-based neuroimaging support package that is unbiased towards any formatting standards and provides 130 applications, more than that of NeuroDebian (51), BIDS App (45), Flywheel (70), and Neurodesk (85). Using a technology stack that employs GUI as the front-end and shell scripted back-end, aXonica provides (i) 130 tools that span the entire MRI-based neuroimaging analysis, and allow the user to (ii) select the software of their choice, (iii) automatically resolve individual dependencies and (iv) installs them. Hence, aXonica can serve as an important resource for researchers and teachers working in the field of MRI-based Neuroimaging to (a) develop software workstations, and/or (b) install newer tools in their existing workstations.

磁共振成像（MRI）有助于研究神经系统。核磁共振成像扫描需要经过大量处理，才能将最终图像呈现给医疗从业人员。有了出色的软件流水线，这些处理过程才得以轻松执行。然而，建立软件工作站并非易事，需要生命科学研究人员能够自如地下载、安装和编写软件脚本，而这些软件对用户并不友好，可能缺乏基本的图形用户界面。由于研究人员难以掌握这些技能，因此迫切需要开发能够自动安装软件管道的软件包，以加快软件工作站和实验室的建设。以往的解决方案包括 NeuroDebian、BIDS Apps、Flywheel、QMENTA、Boutiques、Brainlife 和 Neurodesk。总体而言，所有这些解决方案相辅相成。NeuroDebian 涵盖神经科学，范围更广，只提供 51 种核磁共振成像工具。而 BIDS Apps 致力于 BIDS 格式，只涵盖 45 种与核磁共振成像相关的软件。Boutiques 更为灵活，便于其管道作为独立容器轻松安装、验证、发布和执行。而 Flywheel 和 Qmenta 都是专有软件，只有 NeuroDebian、Brainlife、Neurodesk 和 BIDS Apps 这四款工具可供用户 "免费使用"。本文对过去十年中发表的 317 种基于 MRI 的神经成像工具进行了广泛调查，并介绍了 "aXonica"，这是一种基于 MRI 的神经成像支持软件包，不偏向任何格式标准，可提供 130 种应用，比 NeuroDebian（51 种）、BIDS App（45 种）、Flywheel（70 种）和 Neurodesk（85 种）多。aXonica 采用图形用户界面作为前端、shell 脚本作为后端的技术堆栈，提供：(i) 130 种工具，涵盖整个基于核磁共振成像的神经成像分析，并允许用户 (ii) 选择自己喜欢的软件，(iii) 自动解决个别依赖关系，(iv) 安装它们。因此，aXonica 可作为从事基于 MRI 的神经成像领域工作的研究人员和教师的重要资源，用于 (a) 开发软件工作站，和/或 (b) 在现有工作站中安装更新的工具。

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引用次数: 0

Antibiotic susceptibility and virulence factors of bacterial species among cancer patients 癌症患者对细菌种类的抗生素敏感性和毒力因素

Biotechnology Notes

Pub Date : 2024-01-01 DOI: 10.1016/j.biotno.2024.02.002

Gamal M. El-Sherbiny , Eman E. Farghal , Mohamed K. Lila , Yousseria M. Shetaia , S.S. Mohamed , Marwa MF. Elswify

Antibiotic resistance is one of the most significant challenges of the 20-s century, and the misuse of antibiotics is a driver of antimicrobial resistance. This study aimed to assess the prevalence of multidrug resistance, and detection of its produce virulence factors, including extended-spectrum β-lactamases (ESβLs), biofilm, and siderophores produced by bacterial species isolated from cancer patients. One hundred and seventy-five Gram-negative bacterial isolates were isolated from different samples collected from cancer patients admitted to the National Cancer Institute (NCI), Cairo, Egypt, and processed by standard microbiological methods. One hundred and forty-three bacterial isolates were recovered from adult patients, and 32 were recovered from children. Escherichia coli showed the highest frequency (36%), followed by Klebsiella pneumonia (30.85%), Acinetobacter baummannii (14.28%), and Pseudomonas sp. (9.14%). Antibiotic profiles revealed that bacterial isolates are highly resistant to the most commonly available antibiotics. Amikacin and gentamicin were the most effective antibiotics against isolated Gram-negative bacteria. Moreover, the vast majority of bacterial stains produce virulence factors, including EsβLs, biofilm, and siderophores. E. coli isolates produced ESβLs with rates of 25.28%, Klebsiella pneumonia (11.0%), and Pseudomonas sp. (25.0%). Among these collected bacterial isolates, 132 (75.4%) have the ability to form a biofilm to different degrees. Also, the majority of the bacteria isolates generated siderophores, with 133 (75.94%). This study revealed that a significant distribution of multidrug-resistant pathogenic bacteria may increase the burden on healthcare to prevent infections in cancer patients.

抗生素耐药性是 20 世纪最重大的挑战之一，而滥用抗生素是抗生素耐药性的驱动因素。本研究旨在评估多重耐药性的流行情况，并检测其产生的毒力因子，包括从癌症患者体内分离出的细菌物种所产生的广谱β-内酰胺酶（ESβLs）、生物膜和嗜硒酸盐。从埃及开罗国家癌症研究所（NCI）收治的癌症患者的不同样本中分离出 175 株革兰氏阴性细菌，并采用标准微生物学方法进行处理。其中 143 株细菌分离物来自成年患者，32 株来自儿童。大肠埃希菌出现频率最高（36%），其次是肺炎克雷伯菌（30.85%）、鲍曼不动杆菌（14.28%）和假单胞菌（9.14%）。抗生素谱显示，细菌分离物对最常用的抗生素高度耐药。阿米卡星和庆大霉素是对分离出的革兰氏阴性细菌最有效的抗生素。此外，绝大多数细菌菌斑都会产生毒力因子，包括 EsβLs、生物膜和嗜苷酸盐。大肠杆菌（25.28%）、肺炎克雷伯菌（11.0%）和假单胞菌（25.0%）产生 ESβL。在收集到的细菌分离物中，有 132 种（75.4%）具有不同程度的形成生物膜的能力。此外，大多数细菌分离物都能产生嗜苷酸盐，共有 133 种（75.94%）。这项研究表明，具有多重耐药性的病原菌的大量分布可能会增加医护人员预防癌症患者感染的负担。

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引用次数: 0

A comparative exploration of mRNA capping enzymes mRNA 封口酶的比较研究

Biotechnology Notes

Pub Date : 2024-01-01 DOI: 10.1016/j.biotno.2024.11.005

Yiming Wang , Xiaoxue Wang , Wenchao Li , Xinjie Chen , Yuan Lu

With the wide application of messenger RNA (mRNA) technology in medicine and vaccine fields, higher requirements are put forward for mRNA expression efficiency in vivo. Since the 5′ cap structure can spatially protect mRNA from exonuclease degradation and enhance the initiation of translation reactions, in vitro mRNA caps are a promising option to improve the efficiency of mRNA expression in vivo. In order to obtain more efficient mRNA capping enzymes, seven mRNA capping enzymes from different viral sources were explored in this study. Eukaryotic and prokaryotic cells were used for the heterologous expression of the cap enzymes, and Escherichia coli was identified as the most suitable host cell for heterologous expression. In addition, in order to improve the solubility of the capping enzyme, four kinds of soluble labels were screened, among which maltose-binding protein had the best effect and the widest applicability. The mRNA was then transfected into the human cells, and the highest transfection efficiency was achieved using the bluetongue virus capping enzyme. Its effect was 38 % higher than that of the previously widely used vaccinia virus capping enzyme. This work will promote the development of mRNA technology and expand its application space.

随着信使核糖核酸（mRNA）技术在医药和疫苗领域的广泛应用，对 mRNA 在体内的表达效率提出了更高的要求。由于5′帽结构可以在空间上保护mRNA不被外切酶降解，并增强翻译反应的启动，因此体外mRNA帽是提高体内mRNA表达效率的一个很有前景的选择。为了获得更高效的 mRNA 加帽酶，本研究探索了来自不同病毒源的七种 mRNA 加帽酶。真核细胞和原核细胞被用来异源表达封顶酶，大肠杆菌被确定为最适合异源表达的宿主细胞。此外，为了提高封端酶的溶解度，筛选了四种可溶性标签，其中麦芽糖结合蛋白的效果最好，适用范围最广。然后将 mRNA 转染到人体细胞中，蓝舌病毒封端酶的转染效率最高。其效果比以前广泛使用的疫苗病毒封顶酶高出 38%。这项工作将推动 mRNA 技术的发展，拓展其应用空间。

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