Neoechinulin A as a Promising SARS-CoV-2 Mpro Inhibitor: In Vitro and In Silico Study Showing the Ability of Simulations in Discerning Active from Inactive Enzyme Inhibitors

The COVID-19 pandemic and its continuing emerging variants emphasize the need to discover appropriate treatment, where vaccines alone have failed to show complete protection against the new variants of the virus. Therefore, treatment of the infected cases is critical. This paper discusses the bio-guided isolation of three indole diketopiperazine alkaloids, neoechinulin A (1), echinulin (2), and eurocristatine (3), from the Red Sea-derived Aspergillus fumigatus MR2012. Neoechinulin A (1) exhibited a potent inhibitory effect against SARS-CoV-2 M^pro with IC₅₀ value of 0.47 μM, which is comparable to the reference standard GC376. Despite the structural similarity between the three compounds, only 1 showed a promising effect. The mechanism of inhibition is discussed in light of a series of extensive molecular docking, classical and steered molecular dynamics simulation experiments. This paper sheds light on indole diketopiperazine alkaloids as a potential structural motif against SARS-CoV-2 M^pro. Additionally, it highlights the potential of different molecular docking and molecular dynamics simulation approaches in the discrimination between active and inactive structurally related M^pro inhibitors.     

First molecular detection of canine herpesvirus 1 (CaHV-1) in the Eastern Brazilian Amazon

BackgroundCanine herpesvirus type 1 (CaHV-1) infects dogs and is associated with neonatal deaths and reproductive, ocular, neurological, and respiratory problems. In Brazil, reports of CaHV-1 have been restricted to the southeast and south regions, particularly in municipalities in the state of Rio Grande do Sul.ObjectivesTo assess the presence and variability of CaHV-1 in canine populations in the state of Pará, North Brazil.MethodsBiological samples from 159 dogs from 4 municipalities in the State of Pará were evaluated using polymerase chain reaction and phylogenetic analyses, with the target being the viral enzyme, thymidine kinase.ResultsCaHV-1 was detected in 13 dogs (8.2%), with 2 animals being from the municipality of Santa Bárbara do Pará, 8 from Algodoal Island, 2 from Salinópolis, and one from Capanema. The study sequences revealed 100% identity among themselves and 64% to 100% identity with the other nucleotide sequences from Australia, Brazil, United Kingdom, and United States, including 100% identity with the 2002 isolate from Australia. The 1996 isolate from France was grouped in a branch that was different from the sequence of this study.ConclusionsThis study presents the first molecular detection of CaHV-1 in dogs from the Amazon region in northern Brazil. The nucleotide identity between the strains and cytosine insertion in the sequences isolated in this study suggests at least 2 strains of CaHV-1 circulating in Brazil (Pará and BTU-1).     

苹果杂交后代植株果实着色性状的遗传分析和预测

苹果着色问题是影响我国许多产区苹果质量的重要因素,本研究利用苹果着色相关基因MdMYB1（GenBank登录号：DQ886414）设计引物进行PCR扩增,开发出一个酶切位点为HaeⅢ的CAPS标记Mb2,该标记可用于区分杂交亲本‘富士’和‘嘎拉’苹果及分析其杂交后代植株。进一步利用Mb2标记对该杂交组合的后代植株进行遗传分析,结果表明,可将77个后代植株分为2组,一组42个后代植株含有来自‘嘎拉’亲本的非红色性状等位基因以及来自‘富士’亲本的非红色性状等位基因或者红色性状等位基因,其果实颜色预测为非红色与红色或者非红色与桔红色;另一组35个后代植株含有来自‘嘎拉’亲本的红色性状等位基因以及来自‘富士’亲本的非红色性状等位基因或者红色性状等位基因,其果实颜色预测为桔红色与红色或者只有红色。本研究结果将为选育优质苹果新品种提供依据和方法参考。     

猪CD8B表达谱、基因多态性及其与血液免疫性状的关联分析

白细胞分化抗原8(cluster of differentiation 8,CD8)是主要表达在CD8+T细胞表面的共受体和信号转导分子.CD8B基因编码CD8蛋白的β链,在免疫应答中起着潜在的调节作用.为分析CD8B基因在猪(Sus scrofa)不同组织中的表达特征,并探究CD8B基因多态性对血液免疫性状的遗传效应,本研究利用qRT-PCR测定CD8B基因在大白猪7个组织中的表达量,并用PCR产物测序与限制性片段长度多态性(restriction fragment length polymorphism,RFLP)方法对382头大白猪、84头长白猪和90头松辽黑猪CD8B基因编码区进行SNPs检测,以及分析CD8B基因突变对大白猪外周血T淋巴细胞亚群和血常规指标的影响.结果表明,CD8B基因在脾脏中的mRNA表达量最高,然后依次是肺脏、胃、肝脏、肾脏、心脏和肌肉.CD8B基因外显子5处检测到1个错义突变(c.602G＞A),在大白猪、长白猪和松辽黑猪群中均存在3种基因型(AA、AG和GG型),且3个猪种都是GG为优势基因型.统计分析显示,CD8B基因多态性与大白猪血液中的CD4+CD8-、CD4 CD8+、CD4 +/CD8+、血红蛋白含量(hemoglobin,HGB)和红细胞压积(hematocrit,HCT)指标显著相关(P＜0.05).最小二乘分析表明,3种CD8B基因型个体具有不同的血液参数,其中GG型的CD4+CD8-指标显著高于AG型,AG型的CD4-CD8+指标显著高于GG和AA型,GG和AA型的CD4+/CD8+、HGB和HCT指标显著高于AG型(P＜0.05).因此,CD8B基因可能参与血液生理指标的调控,可作为影响猪免疫性状的重要功能候选基因.本研究为进一步揭示CD8B基因的生物学功能提供依据,也为猪的标记辅助抗病育种提供了基础资料.     

绵羊KAP1.1基因多态性与毛纤维直径的相关性分析

以768只中国美利奴羊(新疆军垦型)为材料,采用测序和PCR技术分析KAP1.1(B2A)基因单核苷酸多态性及其与羊毛纤维直径的相关性。结果表明:中国美利奴羊(新疆军垦型)KAP1.1基因存在30个碱基的插入/缺失和6种基因型,分别为(341 bp)AA基因型、(311 bp)BB基因型、(281 bp)CC基因型、(341 bp/311 bp)AB基因型、(341 bp/281 bp)AC基因型和(311 bp/281 bp)BC基因型。AA、BC、AC、AB和CC基因型间的毛卷曲度、细度离散、毛长、污毛重、净毛率和密度差异不显著(P>0.05),而BB基因型羊只的平均纤维直径显著小于AA、BC、AC、AB和CC基因型羊只的平均纤维直径(P<0.05)。以上结果表明,KAP1.1基因可作为绵羊毛纤维直径的主要候选基因之一,BB基因型可作为分子标记用于预测绵羊毛纤维直径。     

无核葡萄种质亲缘关系的SRAP研究

为充分了解及更合理地利用无核葡萄的种质资源,辅助无核葡萄育种,本研究采用SRAP分子标记技术,对23 个国内外主流无核葡萄品种的种质亲缘关系进行分析。结果表明：从45 对SRAP引物中筛选出38 对多态性引物,共扩增出236 条谱带,其中171 条为多态性谱带,多态性谱带百分率为72.4%。遗传相似分析显示,供试材料间遗传相似系数的变化范围为0.58~0.90,遗传距离在1.0313~4.9643 范围内变化。在遗传相似系数为0.61 处,可将供试材料分为2 个大类群,在相似系数为0.66 处,Ⅰ类群和Ⅱ类群均可分为2个亚群。说明23个品种总体来讲亲缘关系较远,特别欧美杂种具有较高的遗传多样性,部分欧亚种品种间亲缘关系较近,杂交育种时应注意亲本选择。     

棉花杂交种SSR核心引物的筛选与评价

 为筛选到一套针对棉花杂交种真实性和纯度进行鉴定的核心引物,本实验以79个棉花杂交种为材料,采用分子标记技术,对已公布的2300对SSR引物通过PCR扩增,经初筛和复选,最终获得扩增条带清晰、重复性好且分辨能力强的56对核心引物。利用这56对SSR引物对生产上推广的37个棉花杂交种进行多态性验证,共检测到95个多态性位点,变幅为1～3个;161个基因型,变幅为2～3个。随机抽出10对引物,通过组合方式可以将37个品种完全区分开。对37个品种的SSR分子标记结果进行UPGMA聚类,在相似系数为0.54时, 20个长江流域杂交种中有19个被划分到第一类,17个黄河流域品种中有11个被划分到第二类,较好地反映出不同棉区因育种目标的不同所产生的差异。表明该套引物有很好的代表性、实用性和有效性。     

白三叶草一种叶斑病病原菌的分离鉴定

明确在山东省聊城市白三叶草上发现的一种叶斑病的病原菌,以期为该病害的防控提供参考.2019年采集山东省聊城市东昌府区具有叶斑病的白三叶叶片,利用组织分离法分离病原菌,并依据柯赫式法则验证致病性.在形态学观察的基础上,对病原菌进行分子生物学鉴定,构建系统发育树,分析其遗传多样性.从发病叶片中分离得到病原菌(标记为SY1)...     

医药分子农业的受体系统评述

利用转基因植物以农业规模生产口服疫苗、植物抗体、药用蛋白的途径称为医药分子农业。随着医药分子农业的发展,可作为受体的植物的种类正逐步扩大。但对于一个特定的目标蛋白,选用何种受体植物更有利于外源蛋白的高效稳定表达和经济快捷的提纯,仍是需要首要考虑和解决的问题。本文结合最近的研究进展,阐述各类植物受体的特点及应用情况,以期为科学地确定医药分子农业的适宜植物受体系统提供参考。