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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
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
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
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
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
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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引用次数: 0
Screening, characterization, and production of Bacillus cereus (S55) bioflocculant isolated from soil for application in wastewater treatment 筛选、鉴定和生产从土壤中分离出来的蜡样芽孢杆菌(S55)生物絮凝剂,用于废水处理
Pub Date : 2024-01-01 DOI: 10.1016/j.biotno.2024.11.003
Karthikeyan Harinisri , Ragothaman Prathiviraj , Balasubramanian Thamarai Selvi
This research evaluated a bacterial strain, Bacillus cereus (S55), isolated from the soil for its bioflocculant production capabilities. The strain was identified through morphological and 16s rRNA gene sequencing. The optimization of culture conditions, using One-factor-at-a-time method, significantly enhanced bioflocculant production with glucose and urea. FTIR analysis showed the presence of hydroxyl, amine and carboxylate functional groups, with polysaccharides as predominant components. Scanning electron microscopy and X-ray diffraction confirmed the crystalline nature of the bioflocculant. The strain studied showed potential in treating household wastewater and was effective at removing dyes, suggesting alternatives for wastewater management.
这项研究评估了从土壤中分离出来的蜡样芽孢杆菌(S55)的生物絮凝剂生产能力。通过形态学和 16s rRNA 基因测序鉴定了该菌株。采用 "一次一因素 "法优化培养条件,显著提高了葡萄糖和尿素的生物絮凝剂产量。傅立叶变换红外光谱分析显示了羟基、胺和羧基官能团的存在,多糖是主要成分。扫描电子显微镜和 X 射线衍射证实了生物絮凝剂的结晶性质。所研究的菌株显示出处理家庭废水的潜力,并能有效去除染料,为废水管理提供了替代方案。
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引用次数: 0
Organ-on-chip technology: Opportunities and challenges 片上器官技术:机遇与挑战
Pub Date : 2024-01-01 DOI: 10.1016/j.biotno.2024.01.001
Santosh Kumar Srivastava , Guo Wei Foo , Nikhil Aggarwal , Matthew Wook Chang

Organ-on-chip (OOC) technology is an innovative approach that reproduces human organ structures and functions on microfluidic platforms, offering detailed insights into intricate physiological processes. This technology provides unique advantages over conventional in vitro and in vivo models and thus has the potential to become the new standard for biomedical research and drug screening. In this mini-review, we compare OOCs with conventional models, highlighting their differences, and present several applications of OOCs in biomedical research. Additionally, we highlight advancements in OOC technology, particularly in developing multiorgan systems, and discuss the challenges and future directions of this field.

芯片上器官(OOC)技术是一种在微流控平台上再现人体器官结构和功能的创新方法,可让人们详细了解复杂的生理过程。与传统的体外和体内模型相比,该技术具有独特的优势,因此有可能成为生物医学研究和药物筛选的新标准。在这篇微型综述中,我们比较了 OOC 与传统模型,强调了它们之间的差异,并介绍了 OOC 在生物医学研究中的几种应用。此外,我们还强调了 OOC 技术的进步,尤其是在开发多器官系统方面,并讨论了这一领域面临的挑战和未来的发展方向。
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引用次数: 0
Understanding virus retention mechanisms on protein a chromatography based on using different wash buffers – Evaluating the possibility for a generic wash buffer toolbox to improve virus clearance capacity 通过使用不同的清洗缓冲液,了解病毒在蛋白 a 层析上的滞留机制 - 评估通用清洗缓冲液工具箱的可能性,以提高病毒清除能力
Pub Date : 2024-01-01 DOI: 10.1016/j.biotno.2024.03.001
Sandra Krause , Florian Capito , Verena Oeinck , Hendrik Flato , Holger Hoffmann , Ozan Ötes , Annette Berg

During manufacturing of mammalian-cell derived monoclonal antibodies (mAbs) virus clearance capacity of the downstream process has to be demonstrated. The protein A chromatography step typically achieves less than 4 log10 and is not considered as a major contributing step. Having been successfully applied to host cell protein removal before, we used different wash buffers for three mAbs with two model viruses (Minute virus of mice and Murine leukemia virus) in series as well as separately to further understand major contributing interactions for virus retention and potentially design a generic toolbox of stringent wash buffers to be applied to various mAbs. Results indicate a major relevance of hydrophobic interaction for Murine leukemia virus (xMuLV) and mAb A, based on improved clearance for buffers additionally containing increased levels of hydrophobic compounds. This effect was less pronounced for Minute virus of mice (MVM), whereby hydrogen-bonds were expected to play a stronger role for this model virus. Additionally, electrostatic interactions presumably are more relevant for MVM retention compared to xMuLV under the conditions evaluated. A generic mAb and virus-independent stringent wash buffer toolbox could not be identified. However, based on our results a customized mAb and virus wash buffer design with improved virus clearance is possible, with here demonstrated log reduction increase by 1.3 log10 for MVM and 2.2 log10 for xMuLV for the protein A step compared to equilibration buffer alone.

在生产源自哺乳动物细胞的单克隆抗体(mAbs)时,必须证明下游工艺的病毒清除能力。蛋白 A 层析步骤通常能达到小于 4 log10 的清除率,因此不被认为是主要的清除步骤。在成功应用于宿主细胞蛋白清除之前,我们对三种 mAbs 和两种模型病毒(小鼠细小病毒和鼠白血病病毒)分别使用了不同的水洗缓冲液,以进一步了解导致病毒滞留的主要相互作用,并有可能设计出适用于各种 mAbs 的通用严格水洗缓冲液工具箱。结果表明,疏水相互作用对鼠白血病病毒(xMuLV)和 mAb A 有重大意义,因为含有更多疏水化合物的缓冲液的清除率提高了。这种效应在小鼠细小病毒(MVM)中并不明显,因为氢键在这种模式病毒中的作用预计会更大。此外,在评估条件下,与 xMuLV 相比,静电相互作用对 MVM 的保留作用可能更大。我们无法找到通用的 mAb 和病毒无关的严格洗涤缓冲液工具箱。不过,根据我们的研究结果,定制的 mAb 和病毒清洗缓冲液设计可以提高病毒清除率,与单独的平衡缓冲液相比,MVM 和 xMuLV 在蛋白 A 步骤中的对数值分别降低了 1.3 log10 和 2.2 log10。
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引用次数: 0
Incorporating omics-based tools into endophytic fungal research 将基于omics的工具纳入内生真菌研究
Pub Date : 2024-01-01 DOI: 10.1016/j.biotno.2023.12.006
Vinita Verma , Alok Srivastava , Sanjay Kumar Garg , Vijay Pal Singh , Pankaj Kumar Arora

Fungal endophytes are valuable sources of bioactive compounds with diverse applications. The exploration of these compounds not only contributes to our understanding of ecological interactions but also holds promise for the development of novel products with agricultural, medicinal, and industrial significance. Continued exploration of fungal endophyte diversity and understanding the ecological roles of bioactive compounds present opportunities for new discoveries and applications. Omics techniques, which include genomics, transcriptomics, proteomics, and metabolomics, contribute to the discovery of novel bioactive compounds produced by fungal endophytes with their potential applications. The omics techniques play a critical role in unraveling the complex interactions between fungal endophytes and their host plants, providing valuable insights into the molecular mechanisms and potential applications of these relationships. This review provides an overview of how omics techniques contribute to the study of fungal endophytes.

真菌内生菌是具有多种用途的生物活性化合物的宝贵来源。对这些化合物的探索不仅有助于我们了解生态相互作用,而且有望开发出具有农业、医药和工业意义的新型产品。继续探索真菌内生菌的多样性和了解生物活性化合物的生态作用为新发现和新应用提供了机会。包括基因组学、转录物组学、蛋白质组学和代谢组学在内的 Omics 技术有助于发现真菌内生菌产生的新型生物活性化合物及其潜在应用。omics 技术在揭示真菌内生菌与其寄主植物之间复杂的相互作用方面发挥着至关重要的作用,为了解这些关系的分子机制和潜在应用提供了宝贵的见解。本综述概述了全元素技术如何促进真菌内生菌的研究。
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引用次数: 0
The role of brain-derived neurotrophic factor and its recombinant pro-isoforms in depressive disorder: Open questions 脑源性神经营养因子及其重组原异构体在抑郁障碍中的作用:悬而未决的问题
Pub Date : 2024-01-01 DOI: 10.1016/j.biotno.2024.09.001
Éllen F. Rodrigues , Ana L. Fachin , Mozart Marins , Felipe Britto Letieri , Rodrigo G. Stabeli , Renê O. Beleboni
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引用次数: 0
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