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In Silico Design and Evaluation of a Novel Therapeutic Agent Against the Spike Protein as a Novel Treatment Strategy for COVID-19 Treatment. 针对尖峰蛋白的新型治疗剂的硅学设计与评估,作为治疗 COVID-19 的新型治疗策略。
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-01-01 DOI: 10.2174/1872208317666230523105759
Soroush Sarmadi, Mohammad Reza Rahbar, Hamideh Najafi, Onyeka S Chukwudozie, Mohammad Hossein Morowvat

Background: Coronavirus disease 2019 (COVID-19) is a viral respiratory disease that is associated with severe damage to other human organs. It causes by a novel coronavirus, and it is spreading all over the world. To date, there is some approved vaccine or therapeutic agent which could be effective against this disease. But their effectiveness against mutated strains is not studied completely. The spike glycoprotein on the surface of the coronaviruses gives the virus the ability to bind to host cell receptors and enter cells. Inhibition of attachment of these spikes can lead to virus neutralization by inhibiting viral entrance.

Aims: In this study, we tried to use the virus entrance strategy against itself by utilizing virus receptor (ACE-2) in order to design an engineered protein consisting of a human Fc antibody fragment and a part of ACE-2, which reacts with virus RBD, and we also evaluated this interaction by computational methods and in silico methods. Subsequently, we have designed a new protein structure to bind with this site and inhibit the virus from attaching to its cell receptor, mechanically or chemically.

Methods: Various in silico software, bioinformatics, and patent databases were used to retrieve the requested gene and protein sequences. The physicochemical properties and possibility of allergenicity were also examined. Three-dimensional structure prediction and molecular docking were also performed to develop the most suitable therapeutic protein.

Results: The designed protein consisted of a total of 256 amino acids with a molecular weight of 28984.62 and 5.92 as a theoretical isoelectric point. Instability and aliphatic index and grand average of hydropathicity are 49.99, 69.57 and -0.594, respectively.

Conclusions: In silico studies can provide a good opportunity to study viral proteins and new drugs or compounds since they do not need direct exposure to infectious agents or equipped laboratories. The suggested therapeutic agent should be further characterized in vitro and in vivo.

背景:冠状病毒病 2019(COVID-19)是一种病毒性呼吸道疾病,会对人体其他器官造成严重损害。它由一种新型冠状病毒引起,目前正在全球蔓延。迄今为止,有一些已获批准的疫苗或治疗剂可以有效预防这种疾病。但它们对变异毒株的有效性还没有完全研究清楚。冠状病毒表面的尖头糖蛋白使病毒能够与宿主细胞受体结合并进入细胞。目的:在这项研究中,我们试图利用病毒受体(ACE-2)来对抗病毒的进入策略,从而设计出一种由人类 Fc 抗体片段和 ACE-2 的一部分组成的工程蛋白,它能与病毒 RBD 发生反应,我们还通过计算方法和硅学方法对这种相互作用进行了评估。随后,我们设计了一种新的蛋白质结构,与该位点结合,通过机械或化学方法抑制病毒附着到细胞受体上:方法:我们使用了各种硅学软件、生物信息学和专利数据库来检索所需的基因和蛋白质序列。此外,还研究了理化性质和过敏性的可能性。此外,还进行了三维结构预测和分子对接,以开发最合适的治疗蛋白:所设计的蛋白质由 256 个氨基酸组成,分子量为 28984.62,理论等电点为 5.92。不稳定性和脂肪指数及水合平均值分别为 49.99、69.57 和 -0.594:硅学研究为研究病毒蛋白质和新药物或化合物提供了一个良好的机会,因为它们不需要直接接触传染性病原体,也不需要配备实验室。建议的治疗药物应在体外和体内进一步表征。
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引用次数: 0
Botanical Extracts and Compounds of Castanea Plants and Methods of Use: US20190125818A1 - The United States Patent Evaluation. 栗属植物的植物提取物和化合物及其使用方法:US20190125818A1-美国专利评估
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-01-01 DOI: 10.2174/1872208317666230420105000
Tatiane Batista Dos Santos, Denilson Dos Santos Gomes, Agenor Gomes Dos Santos Neto, Lívia Maria do Amorim Costa Gaspar, Daniela Droppa-Almeida

Background: Bacterial infections are increasingly difficult to combat, which makes them a threat to public health on a global level. Staphylococcus aureus is considered one of the main causes of infections in hospitals, as it has a variety of virulence factors, as well as is able to produce bacterial biofilms, which, consequently, bring numerous damages to public health as a result of increased resistance to conventional antibiotics and a longer hospital stay. Therefore, the use of compounds extracted from medicinal plants is a potential pharmaceutically acceptable target, as they do not have toxicity and the potential to disrupt biofilms produced by Staphylococcus aureus already evidenced, thus revealing their relevance to our study.

Objective: The objective of this work was to perform a critical analysis of a patent with natural extracts against bacterial biofilms found in the United States Patent and Trademark Office (USPTO) database, to map the possible bioactive compounds that may serve as potential future antimicrobial drugs.

Methods: A technological survey was carried out to verify existing patents using natural extracts with anti-biofilm potential. For this, it was searched with the keywords: Botanical extracts AND biofilms; which were performed in the United States Patent and Trademark Office (USPTO) database. Thus, the selected patent used a non-aqueous extract partitioned and vacuum-contracted, subsequently lyophilized for assays with antimicrobial potential. Because of this, a patent was analyzed regarding its chemistry, and biological activity, followed by a critical analysis of the technology proposed in the invention.

Results: When using the keywords Botanical extracts AND biofilms in the USPTO, it was possible to find twenty-two inventions; however, only four patents in the USPTO were in agreement with the proposal of the natural extract having antimicrobial activity and an anti-biofilm potential, of which two belonged to the same applicant with similar proposals. The key point of this invention was to enable the compounds of the Castanea sativa plant and its methods of obtaining the extract to present a significant antimicrobial action associated or not with antibiotics, promoting the development of new therapies against bacterial infections capable of disrupting biofilms. The invention developed a methodology for extracting Castanea sativa, in which pentacyclic triterpene compounds were found mostly in its leaves. Whereas for the extraction, the crude methanol extracts called extracts 224 from the ground leaves were made by maceration, filtered, combined, concentrated under pressure in rotary evaporators, and lyophilized. After that, they were resuspended in water and partitioned in succession with hexane, ethyl acetate, and butanol. The most active refined partition was the 224C extract with the solvent ethyl ace

细菌感染越来越难以对抗,这使它们对全球公共健康构成威胁。金黄色葡萄球菌被认为是医院感染的主要原因之一,因为它具有多种毒力因子,并且能够产生细菌生物膜,因此,由于对传统抗生素的耐药性增加和住院时间延长,细菌生物膜会给公众健康带来许多损害。因此,使用从药用植物中提取的化合物是一个潜在的药学可接受的靶点,因为它们没有毒性,并且有可能破坏金黄色葡萄球菌产生的生物膜,这已经证明了它们与我们的研究的相关性。这项工作的目的是对美国专利商标局(USPTO)数据库中发现的一项具有天然提取物的专利进行批判性分析,以绘制可能成为未来潜在抗菌药物的生物活性化合物。进行了一项技术调查,以验证使用具有抗生物膜潜力的天然提取物的现有专利。为此,搜索关键词为:植物提取物和生物膜;其在美国专利商标局(USPTO)数据库中执行。因此,所选专利使用非水提取物进行分配和真空收缩,随后冻干,用于具有抗菌潜力的测定。因此,对专利的化学和生物活性进行了分析,然后对本发明中提出的技术进行了批判性分析。当在USPTO中使用关键词植物提取物和生物膜时,可以发现二十二项发明;然而,USPTO中只有四项专利与具有抗菌活性和抗生物膜潜力的天然提取物的提案一致,其中两项属于具有类似提案的同一申请人。本发明的关键点是使栗属植物的化合物及其获得提取物的方法能够表现出与抗生素相关或不与抗生素相关的显著抗微生物作用,从而促进针对能够破坏生物膜的细菌感染的新疗法的开发。本发明开发了一种提取锥栗的方法,其中五环三萜化合物主要存在于其叶片中。而对于提取,通过浸渍、过滤、合并、在旋转蒸发器中在压力下浓缩并冷冻干燥从磨碎的叶子中提取的粗甲醇提取物(称为提取物224)。之后,将它们重悬于水中,并依次用己烷、乙酸乙酯和丁醇分配。最活跃的精制分配是用溶剂乙酸乙酯提取的224C提取物,使用硅胶柱色谱对其进行进一步分级。产生了最精细的提取物224C-F2,能够直接作用于细菌(主要是金黄色葡萄球菌)的群体感应,阻断RNAIII的翻译,包括一系列外毒素。关于对金黄色葡萄球菌的抗菌能力,其最低抑菌浓度(MIC)和最低杀菌浓度(MBC)分别为1.56µg/mL-1和>100μg/mL-1。鉴于所分析的专利,有可能验证替代品对减少细菌生物膜影响的重要性,细菌生物膜会对整个行业和健康造成损害。由此,所分析的本发明有一个很有前途的方案,具有抗菌潜力,重点是细菌生物膜的巨大影响。因此,具有抗生物膜潜力的天然提取物有助于最大限度地减少这些具有不同毒力机制的耐多药微生物对健康造成的经济损失。
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引用次数: 0
Innovation and Patenting Activities During COVID-19 and Advancement of Biochemical and Molecular Diagnosis in the Post- COVID-19 Era. 新冠肺炎期间的创新和专利活动以及后新冠肺炎时代生化和分子诊断的进展。
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-01-01 DOI: 10.2174/0118722083262217230921042127
Suman Kumar Ray, Sukhes Mukherjee

The COVID-19 pandemic is to escalate globally and acquire new mutations quickly, so accurate diagnostic technologies play a vital role in controlling and understanding the epidemiology of the disease. A plethora of technologies acquires diagnosis of individuals and informs clinical management of COVID. Some important biochemical parameters for COVID diagnosis are the elevation of liver enzymes, creatinine, and nonspecific inflammatory markers such as C-reactive protein (CRP) and Interleukin 6 (IL-6). The main progression predictors are lymphopenia, elevated D-dimer, and hyperferritinemia, although it is also necessary to consider LDH, CPK, and troponin in the marker panel of diagnosis. Owing to the greater sensitivity and accuracy, molecular technologies such as conventional polymerase chain reaction (PCR), reverse transcription (RT)-PCR, nested PCR, loop-mediated isothermal amplification (LAMP), and xMAP technology have been extensively used for COVID diagnosis for some time now. To make so many diagnostics accessible to general people, many techniques may be exploited, including point of care (POC), also called bedside testing, which is developing as a portable promising tool in pathogen identification. Some other lateral flow assay (LFA)-centered techniques like SHERLOCK, CRISPR-Cas12a (AIOD-CRISPR), and FNCAS9 editor limited uniform detection assay (FELUDA), etc. have shown auspicious results in the rapid detection of pathogens. More recently, low-cost sequencing and advancements in big data management have resulted in a slow but steady rise of next-generation sequencing (NGS)-based approaches for diagnosis that have potential relevance for clinical purposes and may pave the way toward a better future. Due to the COVID-19 pandemic, various institutions provided free, specialized websites and tools to promote research and access to critically needed advanced solutions by alleviating research and analysis of data within a substantial body of scientific and patent literature regarding biochemical and molecular diagnosis published since January 2020. This circumstance is unquestionably unique and difficult for anyone using patent information to find pertinent disclosures at a specific date in a trustworthy manner.

新冠肺炎大流行将在全球范围内升级并迅速获得新的突变,因此准确的诊断技术在控制和了解该疾病的流行病学方面发挥着至关重要的作用。过多的技术可以获得对个人的诊断,并为新冠肺炎的临床管理提供信息。诊断新冠肺炎的一些重要生化参数是肝酶、肌酸酐和非特异性炎症标志物(如C反应蛋白(CRP)和白细胞介素6(IL-6))的升高。主要的进展预测因素是淋巴细胞减少症、D-二聚体升高和高铁蛋白血症,尽管在诊断标志物组中也有必要考虑LDH、CPK和肌钙蛋白。由于其更高的灵敏度和准确性,一段时间以来,常规聚合酶链式反应(PCR)、逆转录(RT)-PCR、嵌套PCR、环介导等温扩增(LAMP)和xMAP技术等分子技术已被广泛用于新冠肺炎诊断。为了让普通人能够获得如此多的诊断,可以利用许多技术,包括护理点(POC),也称为床边检测,它正在发展成为一种便携式的病原体识别工具。其他一些以侧流分析(LFA)为中心的技术,如SHERLOCK、CRISPR-Cas12a(AIOD-CRISPR)和FNCAS9编辑有限的均匀检测分析(FELUDA)等,在病原体的快速检测中显示出良好的结果。最近,低成本测序和大数据管理的进步导致了基于下一代测序(NGS)的诊断方法缓慢但稳定地兴起,这些方法对临床目的具有潜在的相关性,并可能为更美好的未来铺平道路。由于新冠肺炎大流行,各机构提供了免费的专业网站和工具,通过减少自2020年1月以来发表的大量生物化学和分子诊断科学和专利文献中的数据研究和分析,促进研究和获取急需的先进解决方案。毫无疑问,这种情况是独一无二的,任何使用专利信息的人都很难在特定日期以可信的方式找到相关的披露信息。
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引用次数: 0
Development of Moloney Murine Leukemia Virus Reverse Transcriptase Fused with Archaeal DNA-binding Protein Sis7a. 与古DNA结合蛋白Sis7a融合的莫隆尼鼠白血病病毒逆转录酶的开发。
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-01-01 DOI: 10.2174/1872208317666230403104302
Goldyna M Simanjuntak, Azzania Fibriani, Amalia A Fananda, Nicholas Yamahoki

Introduction: Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV RT) is a common enzyme used to convert RNA sequences into cDNA. However, it still has its shortcomings, especially in terms of processivity and thermostability. According to a previous patent, the fusion of polymerase enzyme to an archaeal DNA-binding protein has been proven to enhance its performance. Furthermore, recent studies have also stated that the fusion of a polymerase enzyme to an archaeal DNA-binding protein is predicted to improve its thermostability and processivity.

Aim: As an early stage of enzyme development, this study aimed to design, express, and purify enzymatically active MMLV RT fused with archaeal DNA-binding protein.

Methods: RT fusion proteins were designed and evaluated using in silico methods. The RT fusion enzyme was then expressed in Escherichia coli BL21(DE3) and purified. Its reverse transcriptional activity was proved using reverse transcription quantitative polymerase chain reaction (RT-qPCR).

Results: This study showed that MMLV RT fusion with Sis7a protein at its C-terminal end using commercial linker (GGVDMI) produced the best in silico evaluation results. The RT fusion was successfully expressed and purified. It was also known that the optimal condition for expression of the RT fusion was using 0.5 mM IPTG with post-induction incubation at room temperature (± 26°C) for 16 hours. In addition, the activity assay proved that the RT fusion has the reverse transcriptional activity.

Conclusion: This study shows that the designed MMLV RT Sis7a fusion can be expressed and purified, is enzymatically active, and has the potential to be developed as an improved RT enzyme. Further study is still needed to prove its thermostability and processivity, and further characterize, and plan production scale-up of the MMLV RT Sis7a fusion for commercial use.

简介莫隆尼鼠白血病病毒逆转录酶(MMLV RT)是一种将 RNA 序列转化为 cDNA 的常用酶。然而,它仍有不足之处,尤其是在加工性和热稳定性方面。根据以前的一项专利,聚合酶与古DNA结合蛋白的融合已被证明可提高其性能。目的:作为酶开发的早期阶段,本研究旨在设计、表达和纯化与古DNA结合蛋白融合的具有酶活性的MMLV RT:方法:采用硅学方法设计和评估了 RT 融合蛋白。然后在大肠杆菌 BL21(DE3)中表达并纯化了 RT 融合酶。利用反转录定量聚合酶链反应(RT-qPCR)证明了其反转录活性:研究结果表明,使用商业连接体(GGVDMI)在 MMLV RT 的 C 端与 Sis7a 蛋白融合,产生了最佳的硅学评估结果。该 RT 融合体被成功表达和纯化。研究还发现,表达 RT 融合体的最佳条件是使用 0.5 mM IPTG,诱导后在室温(± 26°C)下培养 16 小时。此外,活性测定也证明了 RT 融合体具有反转录活性:本研究表明,所设计的 MMLV RT Sis7a 融合体可以表达和纯化,具有酶活性,有望开发成一种改良的 RT 酶。仍需进一步研究,以证明其恒温性和加工性,并进一步确定 MMLV RT Sis7a 融合体的特性,计划将其放大生产,用于商业用途。
{"title":"Development of Moloney Murine Leukemia Virus Reverse Transcriptase Fused with Archaeal DNA-binding Protein Sis7a.","authors":"Goldyna M Simanjuntak, Azzania Fibriani, Amalia A Fananda, Nicholas Yamahoki","doi":"10.2174/1872208317666230403104302","DOIUrl":"10.2174/1872208317666230403104302","url":null,"abstract":"<p><strong>Introduction: </strong>Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV RT) is a common enzyme used to convert RNA sequences into cDNA. However, it still has its shortcomings, especially in terms of processivity and thermostability. According to a previous patent, the fusion of polymerase enzyme to an archaeal DNA-binding protein has been proven to enhance its performance. Furthermore, recent studies have also stated that the fusion of a polymerase enzyme to an archaeal DNA-binding protein is predicted to improve its thermostability and processivity.</p><p><strong>Aim: </strong>As an early stage of enzyme development, this study aimed to design, express, and purify enzymatically active MMLV RT fused with archaeal DNA-binding protein.</p><p><strong>Methods: </strong>RT fusion proteins were designed and evaluated using <i>in silico</i> methods. The RT fusion enzyme was then expressed in <i>Escherichia coli</i> BL21(DE3) and purified. Its reverse transcriptional activity was proved using reverse transcription quantitative polymerase chain reaction (RT-qPCR).</p><p><strong>Results: </strong>This study showed that MMLV RT fusion with Sis7a protein at its C-terminal end using commercial linker (GGVDMI) produced the best <i>in silico</i> evaluation results. The RT fusion was successfully expressed and purified. It was also known that the optimal condition for expression of the RT fusion was using 0.5 mM IPTG with post-induction incubation at room temperature (± 26°C) for 16 hours. In addition, the activity assay proved that the RT fusion has the reverse transcriptional activity.</p><p><strong>Conclusion: </strong>This study shows that the designed MMLV RT Sis7a fusion can be expressed and purified, is enzymatically active, and has the potential to be developed as an improved RT enzyme. Further study is still needed to prove its thermostability and processivity, and further characterize, and plan production scale-up of the MMLV RT Sis7a fusion for commercial use.</p>","PeriodicalId":21064,"journal":{"name":"Recent patents on biotechnology","volume":" ","pages":"71-83"},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9254174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Direct Organogenesis of Citrus Cultivars from Shoot Tip Nodal Segments. 柑橘栽培品种的芽尖节段直接器官发生。
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-01-01 DOI: 10.2174/1872208317666230417084141
Zekeria Yusuf, Mulugeta Desta, Wassu Mohammed

Background: Citrus cultivar improvement via conventional breeding strategies is impeded by factors related to its reproductive biology. The orange is a hybrid between pomelo (Citrus maxima) and mandarin (Citrus reticulata). Among various orange cultivars, Valencia oranges have a bit of bitter tang mixed in with their sweetness, as Navel oranges are, the most widely cultivated citrus species, quite sweeter, and also don't contain any seeds. Tangelo mandarin orange cultivar is a hybrid of C. reticulata × C. maxima or × C. paradisi.

Objective: The present study was undertaken to optimize the hormonal composition of the media with regard to plant growth regulators for in vitro propagation of sweet orange cultivars from nodal segment explants.

Methods: The nodal segment explants were collected from three citrus cultivars, Washington Navel, Valencia and Tangelo. Murashige and Skoog (MS) medium supplemented with sucrose and different concentrations of growth regulators was used for shoot proliferation and root induction, and the optimum medium composition was assessed. The patent for Citrus Tissue Culture was obtained from the Office of Research Affairs, Haramaya University.

Results: The results indicate that the highest shoot response was recorded for Washington's navel with maximum shoot proliferation rate (99.75%), shoot number per explant (1.76), shoot length (10.70 cm), leaf number per explants (3.54) after three weeks of culture. In all experiments, no growth was observed for the basal MS medium. Phytohormone combinations of IAA (1.2 mg/L) and kinetin (2.0 mg/L) were found to be the best for shoot proliferation. Among the cultivars, there were significant differences for the highest rooting rate (81.255), root number (2.22), and root length (2.95 cm) variables for Washington Navel. The lowest rooting rate (48.45%), root number (1.47) and root length (2.26 cm) were observed for Valencia. The highest rooting rate (84.90%), root number per microshoot (2.22) and root length (3.05 cm) was on MS medium supplemented with 1.5 mg/L NAA.

Conclusion: A comparison of different concentrations of IAA and NAA on root induction of microshoots from nodal segments of citrus cultivars demonstrated NAA was a more effective hormone than IAA.

背景:通过传统育种策略改良柑橘栽培品种受到其生殖生物学相关因素的阻碍。橙子是柚子(Citrus maxima)和柑橘(Citrus reticulata)的杂交种。在各种橙子栽培品种中,瓦伦西亚橙子的甜味中夹杂着一点苦味,而脐橙是栽培最广泛的柑橘品种,甜度较高,而且不含任何种子。柚子柑栽培品种是 C. reticulata × C. maxima 或 × C. paradisi 的杂交种:本研究旨在优化从节段外植体体外繁殖甜橙栽培品种的培养基中植物生长调节剂的激素组成:方法:从华盛顿脐橙、瓦伦西亚和丹桂三种柑橘栽培品种中采集节段外植体。使用添加了蔗糖和不同浓度生长调节剂的 Murashige 和 Skoog(MS)培养基进行芽增殖和根诱导,并评估了最佳培养基成分。柑橘组织培养的专利权由原谷大学研究事务办公室授予:结果表明,华盛顿脐橙的芽反应最高,培养三周后,芽增殖率(99.75%)、每个外植体的芽数(1.76)、芽长(10.70 厘米)和每个外植体的叶片数(3.54)均达到最高值。在所有实验中,基础 MS 培养基均未观察到生长。IAA(1.2 毫克/升)和松香素(2.0 毫克/升)的植物激素组合对嫩芽增殖效果最好。在各栽培品种中,华盛顿脐橙的最高生根率(81.255)、根数(2.22)和根长(2.95 厘米)变量存在显著差异。瓦伦西亚的生根率(48.45%)、根数(1.47)和根长(2.26 厘米)最低。在添加了 1.5 毫克/升 NAA 的 MS 培养基上,华盛顿脐橙的生根率(84.90%)、每小芽生根数(2.22)和根长(3.05 厘米)最高:比较了不同浓度的 IAA 和 NAA 对柑橘栽培品种节段小芽生根诱导的影响,结果表明 NAA 是比 IAA 更有效的激素。
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引用次数: 0
Comparison of Five Escherichia coli Strains to Achieve the Maximum Yield of a Recombinant Immunotoxin Consisting of an Antibody against VEGF Conjugated with MAP30 Toxin in a Benchtop Bioreactor. 五株大肠杆菌在BenchtopBioreactor中获得由与MAP30毒素结合的VEGF抗体组成的重组免疫毒素的最大产量的比较
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-01-01 DOI: 10.2174/1872208317666230316111554
Mina Zarei, Mohammad Hossein Morowvat

Background: Cancer is among the leading causes of death worldwide, imposing high costs on the health systems of all societies. Extensive biological studies are required to discover appropriate therapies. Escherichia coli has long been regarded as one of the main biotechnological bio-factories to produce recombinant protein-based therapeutics. In the present study, five strains of E. coli were compared to achieve the maximum production of a previously designed recombinant immunotoxin-carrying MAP30 toxin against VEGF-overexpressed cancer cells in a benchtop bioreactor.

Methods: The recombinant immunotoxin coding gene sequence was extracted from the NCBI database. The host used to produce the recombinant immunotoxin were five E. coli strains of BL21 (DE3), DH5α, SHuffle®T7, XL1-Blue, and Rosetta-gamiTM (DE3). CaCl2 method was used for bacterial transformation. Bacterial growth measurements were performed using optical density measurements at 600 nm. The immunotoxin production was measured using SDS-PAGE analysis. The best-producing strain was cultivated in a 10-L benchtop stirred tank bioreactor. Recent patents on this field were also studied.

Results: The results demonstrated that the BL21 (DE3) strain had the highest expression of recombinant protein in comparison to other strains. Moreover, the cell growth of E. coli BL21 (DE3) and SHuffle®T7 strains before transformation in the LB medium, were significantly higher in comparison to other strains. Additionally, the transformation of Rosettagami was associated with decreased cell proliferation. The transformation of the XL1-Blue strain did not effect cell growth. Analysis of the growth kinetics demonstrated appropriate proliferation of the transformed BL21 (DE3) cells in the laboratory benchtop bioreactor.

Conclusions: Based on the results of this study, the BL21 (DE3) strain could be used as a suitable host for the production of the recombinant immunotoxin against VEGF in stirred tank bioreactor, which can be employed for the treatment of tumors. Yet, its precise mechanism must be explored in extensive studies.

癌症是世界范围内的主要死亡原因之一,给所有社会的卫生系统带来了高昂的成本。需要广泛的生物学研究来发现合适的治疗方法。大肠杆菌一直被认为是生产重组蛋白治疗药物的主要生物技术生物工厂之一。在本研究中,研究人员比较了5株大肠杆菌,以在台式生物反应器中最大限度地生产先前设计的携带免疫毒素的重组MAP30毒素,以对抗vegf过表达的癌细胞。从encbi数据库中提取重组免疫毒素编码基因序列。用于产生重组免疫毒素的宿主为BL21 (DE3)、DH5α、SHuffle®T7、XL1-Blue和Rosetta-gamiTM (DE3) 5种大肠杆菌。用cacl2法进行细菌转化。使用600 nm光密度测量进行细菌生长测量。采用SDS-PAGE分析测定免疫毒素的产生。在10- l台式搅拌槽生物反应器中培养出产量最高的菌株。结果表明,与其他菌株相比,BL21 (DE3)菌株的重组蛋白表达量最高。此外,大肠杆菌大肠杆菌21 (DE3)和SHuffle®T7在LB培养基中转化前的细胞生长显著高于其他菌株。此外,Rosetta-gami的转化与细胞增殖减少有关。XL1-Blue菌株的转化对细胞生长没有影响。生长动力学分析表明转化后的BL21 (DE3)细胞在实验室台式生物反应器中有适当的增殖。基于本研究结果,BL21 (DE3)菌株可作为在搅拌槽生物反应器中生产抗VEGF重组免疫毒素的合适宿主,用于肿瘤的治疗。然而,它的确切机制还有待于广泛的研究。基于本研究结果,BL21 (DE3)菌株可作为搅拌式生物反应器中生产抗VEGF重组免疫毒素的合适宿主,用于肿瘤的治疗。然而,它的确切机制还有待于广泛的研究。没有一个
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引用次数: 0
Fundamental Uses of Peptides as a New Model in Both Treatment and Diagnosis. 多肽作为治疗和诊断新模式的基本用途
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-01-01 DOI: 10.2174/1872208317666230512143508
Hend Okasha

An amino acid short chain is known as a peptide. Peptide bonds are the connections that hold the amino acids of a peptide together in a particular order. Characteristically, the shorter length of peptides helps to identify them from proteins. Different ways are used to classify peptides, including chain length, source of peptides, or their biological functions. The fact that peptides serve several purposes suggests that there is a foundation for improvement in peptide production and structure to enhance action. In addition, many patents on peptides for therapeutic and diagnostic approaches have been obtained. This review aims to give an overview of peptides used recently in treatment and diagnosis.

氨基酸短链被称为肽。肽键是将肽的氨基酸按特定顺序连接在一起的连接。从特征上讲,较短长度的肽有助于从蛋白质中识别它们。使用不同的方法对肽进行分类,包括链长、肽来源或其生物功能。肽具有多种用途这一事实表明,肽的生产和结构的改进有助于增强作用。这篇综述旨在概述近年来在治疗和诊断中使用的肽。
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引用次数: 0
Human DNA Mutations and their Impact on Genetic Disorders. 人类DNA突变及其对遗传疾病的影响。
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-01-01 DOI: 10.2174/0118722083255081231020055309
Safia Samir

DNA is a remarkably precise medium for copying and storing biological information. It serves as a design for cellular machinery that permits cells, organs, and even whole organisms to work. The fidelity of DNA replication results from the action of hundreds of genes involved in proofreading and damage repair. All human cells can acquire genetic changes in their DNA all over life. Genetic mutations are changes to the DNA sequence that happen during cell division when the cells make copies of themselves. Mutations in the DNA can cause genetic illnesses such as cancer, or they could help humans better adapt to their environment over time. The endogenous reactive metabolites, therapeutic medicines, and an excess of environmental mutagens, such as UV rays all continuously damage DNA, compromising its integrity. One or more chromosomal alterations and point mutations at a single site (monogenic mutation) including deletions, duplications, and inversions illustrate such DNA mutations. Genetic conditions can occur when an altered gene is inherited from parents, which increases the risk of developing that particular condition, or some gene alterations can happen randomly. Moreover, symptoms of genetic conditions depend on which gene has a mutation. There are many different diseases and conditions caused by mutations. Some of the most common genetic conditions are Alzheimer's disease, some cancers, cystic fibrosis, Down syndrome, and sickle cell disease. Interestingly, scientists find that DNA mutations are more common than formerly thought. This review outlines the main DNA mutations that occur along the human genome and their influence on human health. The subject of patents pertaining to DNA mutations and genetic disorders has been brought up.

DNA是一种非常精确的复制和存储生物信息的介质。它是细胞机械的设计,允许细胞、器官甚至整个生物体工作。DNA复制的保真度源于数百个参与校对和损伤修复的基因的作用。所有的人类细胞都可以在一生中获得DNA的基因变化。基因突变是指在细胞分裂过程中,当细胞复制自己时,DNA序列发生的变化。DNA突变可能会导致遗传疾病,如癌症,或者随着时间的推移,它们可以帮助人类更好地适应环境。内源性反应性代谢产物、治疗药物和过量的环境诱变剂,如紫外线,都会持续损害DNA,损害其完整性。单个位点的一个或多个染色体改变和点突变(单基因突变),包括缺失、重复和反转,说明了这种DNA突变。当基因从父母那里遗传时,可能会发生遗传疾病,这会增加患上这种特殊疾病的风险,或者一些基因改变可能随机发生。此外,遗传疾病的症状取决于哪个基因发生了突变。突变导致了许多不同的疾病和状况。一些最常见的遗传疾病是阿尔茨海默病、一些癌症、囊性纤维化、唐氏综合症和镰状细胞病。有趣的是,科学家们发现DNA突变比以前想象的更常见。这篇综述概述了人类基因组中发生的主要DNA突变及其对人类健康的影响。
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引用次数: 0
Innovative Nanomaterials for Targeting Hypoxia to Improve Treatment for Triple-negative Breast Cancer. 靶向缺氧的创新纳米材料改善三阴性乳腺癌的治疗。
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-01-01 DOI: 10.2174/0118722083270521231027074157
Suman Kumar Ray, Sukhes Mukherjee

Triple-negative breast cancer (TNBC) is an aggressive breast cancer with a high rate of metastases, a short overall survival time, and a poor response to targeted therapy. Improving tumor hypoxia by lowering the oxygen consumption rate of breast tumor cells is a powerful strategy. A viable way to address this issue is to improve therapeutic efficacy by improving the effectiveness of radiation and overcoming drug resistance in TNBC treatment by controlling hypoxia in the tumor microenvironment. The failure of radiation and chemotherapy in TNBC is frequently caused by hypoxia. In TNBC therapy, novel nanomaterials are used for oxygen delivery or generation to affect the tumor microenvironment to improve the effects of ionizing radiation using nanoplatforms. One of the growing fields is novel nano-based drug delivery devices for hypoxic regions and hypoxia- inducible factor-1 (HIF1) targeted therapeutics. Biocompatible nanoparticles may be used in the treatment of TNBC patients in the clinic. Because of the rising market and competition, intellectual property rights (IPR), patents, and tactics may be critically considered. To better comprehend the current state of IPR and patents in cancer nanotechnology, this overview examines recent advances and sophisticated protection measures in this area.

三阴性乳腺癌(TNBC)是一种转移率高、总生存时间短、对靶向治疗反应差的侵袭性乳腺癌。通过降低乳腺肿瘤细胞的耗氧量来改善肿瘤缺氧是一种有效的策略。解决这一问题的可行途径是通过控制肿瘤微环境缺氧,提高放疗效果,克服TNBC治疗中的耐药,从而提高治疗效果。TNBC放化疗失败的原因多为缺氧。在TNBC治疗中,新型纳米材料被用于氧的输送或产生,以影响肿瘤微环境,从而利用纳米平台改善电离辐射的效果。其中一个日益增长的领域是用于缺氧区域和缺氧诱导因子-1 (HIF1)靶向治疗的新型纳米药物递送装置。生物相容性纳米颗粒可用于临床治疗TNBC患者。由于日益激烈的市场和竞争,知识产权(IPR)、专利和策略可能被批判性地考虑。为了更好地了解癌症纳米技术知识产权和专利的现状,本文概述了该领域的最新进展和复杂的保护措施。
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引用次数: 0
An Outline of the Immunogenic Potential of Progressing SARSCoV- 2 Vaccine Technologies among Children and Adolescents. 进展中的SARS-CoV-2疫苗技术在儿童和青少年中的免疫原性潜力概述
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2024-01-01 DOI: 10.2174/1872208317666230612141930
Hytham Ghanem, Shehab Ghanem, Ehsan AlMutawa

Background: SARS-CoV-2, a highly dynamic beta-coronavirus, can afflict all age groups. Notably, over 16100 mortalities have been recorded among children as yet. In this regard, many vaccine projects are operational to assess immuno-potency among young cohorts. A bulk of reports have evidenced the efficacy of these immunization technologies in the elderly population, though the impact is yet to be determined among children.

Objectives: This review is envisioned to outline the current efficacy of contributing vaccine technologies and examine the dose-dependent impact of immunization regimens in lowering the risks of SARS-CoV-2 infections among children and adolescents. Furthermore, the current review exclusively estimated the vaccine impact at current doses.

Methods: A total of 52 research papers extracted from PubMed, Pubmed Central, Science Direct, Research Gate, Google Scholar and Semantic Scholar were screened along with an emphasis on patents. Inclusion criteria involved all published reports directly or indirectly linked to the contributing vaccine candidates that are operational among the young cohort. Unrelated research papers were excluded from the study. Key search terminologies included information on vaccine identifiers, such as name, type and clinical trial ID, and successively restricted to children and adolscents age groups.

Results: Several vaccine designs, such as mRNA-based vaccinations, viral vector vaccines, DNA vaccines, inactivated vaccines, recombinant vaccines, and protein-based immunizations, are being examined at various stages of clinical trials to gauge the effects on children and adolescents. With reference to the published reports, the mRNA 1273 (1610 GMT; 6-10 yrs, 1401 GMT; 12-15 yrs), BNT162b2 (1407 GMT; 6 months- <2 yrs, 1535 GMT; 2-4 yrs, 4583 GMT; 5-11 yrs, 1239.5 GMT; 12-15 yrs) and Ad5 nCoV (1037.5 GMT; 6-17 yrs) offered relatively high neutralization titers with sharp seroconversion rates compared to MVC-COV1901 (648.5 GMT; 12-17 yrs) and ZyCoV-D (133.49 GMT; 12-17 yrs), which produced modest immune responses.

Conclusion: Currently, the WHO is analyzing emerging evidence to issue an emergency use list of vaccines for vaccinating children and adolescents.

SARS-CoV-2是一种高度动态的乙型冠状病毒,可影响所有年龄组。值得注意的是,截至目前,儿童死亡人数已超过16100人。在这方面,许多疫苗项目正在运作,以评估年轻人群体的免疫效力。大量报告证明了这些免疫技术对老年人的有效性,但对儿童的影响尚待确定。本综述旨在概述现有疫苗技术的有效性,并研究免疫方案在降低儿童和青少年感染SARS-CoV-2风险方面的剂量依赖性影响。此外,目前的审查只估计了当前剂量下疫苗的影响。从PubMed、PubMed Central、ScienceDirect、research Gate、谷歌Scholar、Semantic Scholar等网站共筛选了52篇研究论文。纳入标准涉及所有与在年轻队列中起作用的候选疫苗直接或间接相关的已发表报告。不相关的研究论文被排除在研究之外。关键搜索术语包括疫苗标识符信息,如名称、类型和临床试验ID,并先后限制在儿童和青少年年龄组。几种疫苗设计,如基于mrna的疫苗接种、病毒载体疫苗、DNA疫苗、灭活疫苗、重组疫苗和基于蛋白质的免疫接种,正在临床试验的不同阶段进行审查,以评估对儿童和青少年的影响。参考已发表的报告,mRNA 1273(格林威治标准时间1610;6-10年,1401 GMT;12-15年),BNT162b2(格林尼治标准时间1407;6个月- <2年,1535 GMT;2-4年,4583 GMT;5-11年,1239.5 GMT;12-15岁)和Ad5 nCoV(格林威治标准时间1037.5;6-17岁)提供相对较高的中和滴度,与MVC-COV1901 (648.5 GMT;12-17岁)和ZyCoV-D (133.49 GMT;目前,世卫组织正在分析新出现的证据,以发布一份用于为儿童和青少年接种疫苗的紧急使用清单。
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引用次数: 0
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Recent patents on biotechnology
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