革兰氏阴性细菌外膜主要成分的跨膜转运机制研究进展

革兰氏阴性细菌的外膜由脂多糖、磷脂、外膜蛋白和脂蛋白等成分组成,是细菌抵御外界有害物质的首要物理屏障,与细菌致病性和耐药性密切相关.外膜各组分依赖特定的系统进行跨膜转运,包括脂多糖转运系统(lipopolysaccharide transport, Lpt)、脂质不对称维持系统(maintenance of lipid asymmetry, Mla)、β-桶状装配机器(β-barrel assembly machinery,Bam)以及脂蛋白定位系统(localization of lipoprotein,Lol).这些系统能够保证细菌外膜的完整与稳定,被视为维持细菌生命活动的"命门".因此,本文系统地综述革兰氏阴性细菌外膜主要成分的跨膜转运系统结构与功能,并对其未来研究方向进行展望,为新型靶向抗菌类药物研发提供新的思路.     

病原菌调节宿主细胞泛素化途径的研究进展

泛素化是真核生物特有的蛋白质翻译后修饰,广泛地参与宿主细胞各种信号通路和生理过程.病原菌常通过分泌毒性效应蛋白,对泛素和泛素结合酶进行独特的共价修饰,或者利用泛素连接酶和去泛素化酶的酶学活性,调节宿主泛素化过程,从而干扰宿主细胞的信号转导,促进细菌的感染和生存.本文概述了病原菌效应蛋白调节宿主泛素化途径的主要研究进展和最新发现.     

Molecular identification and phylogenetic analysis of fungal pathogens isolated from diseased fish in Xinjiang,China

The outbreaks of fungal diseases in cultured fish have been severe in recent years, which is harmful to the healthy and sustainable development of fish farming. In this study, an investigation was conducted for significant fungal infections of 12 species of fish in four regions in Xinjiang, China, to understand the distribution of local fish fungal pathogens. Twenty-six fungal strains with pathogenicity were isolated, and the challenge experiment showed that eight strains from Changji area had high infection rate to fish eggs. Based on internal transcribed spacer sequence data and molecular analysis, the 26 strains were classified into nine different species of six fungal genera. Phylogenetic analysis showed that all strains were divided into two clades, namely Cluster 1 (contains only the genus Mucor) and Cluster 2 (consists of five small branches), and the distribution of strains from the same region was scattered in two clusters. There is no strict host selectivity for these fungi to infect fish. Mucor sp. are the main fungal pathogen of fish in these four regions, whereas Hypophthalmichthys molitrix and Carassius auratus are two types of fish that were susceptible to pathogen. In addition, the environmental adaptability experiments showed that eight highly pathogenic strains have different adaptability to the environment, and their optimum temperature and pH were 25°C and 7.0, respectively, whereas the concentration of NaCl was negatively correlated with the growth of strains. Therefore, these results indicated that the coinfection of multiple fungal pathogens in a culture region should be considered in the future study.     

盐酸米诺环素软膏辅助龈下刮治术及根面平整术对慢性牙周炎患者龈下牙周致病菌和龈沟液炎性因子的影响

目的:探讨盐酸米诺环素软膏辅助龈下刮治术及根面平整术(FM-SRP)对慢性牙周炎(CP)患者龈下牙周致病菌和龈沟液炎性因子的影响。方法:选择2015年10月到2019年10月期间我院收治的82例CP患者,根据随机数字表法分为对照组(n=41)和观察组(n=41),对照组给予FM-SRP,观察组在对照组基础上联合盐酸米诺环素软膏辅助治疗,比较两组疗效、牙周指标、龈下牙周致病菌和龈沟液炎性因子情况,统计两组不良反应情况。结果:与对照组总有效率70.73%(29/41)相比,观察组治疗后的总有效率90.24%(37/41)更高(P<0.05)。治疗后,两组龈沟出血指数(SBI)、附着水平(AL)、菌斑指数(PLI)、牙周袋深度(PD)均下降,且观察组低于对照组(P<0.05)。治疗后,两组转化生长因子-β(TGF-β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)均下降,且观察组低于对照组(P<0.05)。对比两组不良反应无差异(P>0.05)。治疗后,两组伴防线杆菌、牙龈卟啉单胞菌比例均下降,且观察组低于对照组(P<0.05)。结论:盐酸米诺环素软膏辅助FM-SRP治疗CP患者,可有效消除致病菌,缓解炎性反应,恢复牙周生态平衡,且不增加不良反应发生率,疗效确切。     

Gastrointestinal stress as innate defence against microbial attack

The human gastrointestinal (GI) tract has been bestowed with the most difficult task of protecting the underlying biological compartments from the resident commensal flora and the potential pathogens in transit through the GI tract. It has a unique environment in which several defence tactics are at play while maintaining homeostasis and health. The GI tract shows myriad number of environmental extremes, which includes pH variations, anaerobic conditions, nutrient limitations, elevated osmolarity etc., which puts a check to colonization and growth of nonfriendly microbial strains. The GI tract acts as a highly selective barrier/platform for ingested food and is the primary playground for balance between the resident and uninvited organisms. This review focuses on antimicrobial defense mechanisms of different sections of human GI tract. In addition, the protective mechanisms used by microbes to combat the human GI defence systems are also discussed. The ability to survive this innate defence mechanism determines the capability of probiotic or pathogen strains to confer health benefits or induce clinical events respectively.     

Carbon utilization and growth-inhibition of citrus-colonizing Phyllosticta species

The genus Phyllosticta includes both endophytic and phytopathogenic species that occur on a broad range of plant hosts, including Citrus. Some pathogenic species cause severe disease, such as Phyllosticta citricarpa, the causal agent of Citrus Black Spot (CBS). In contrast, other species, such as Phyllosticta capitalensis, have an endophytic lifestyle in numerous plant hosts. Carbon utilization capabilities are hypothesized to influence both host range and lifestyle, and are in part determined by the set of Carbohydrate Active Enzyme (CAZyme) encoding genes of a species. In this study, carbon utilization capabilities of five Phyllosticta species were determined, as well as the CAZyme repertoire (CAZome) encoded in their genomes. Little variation was found among species in terms of carbon utilization capabilities and CAZome. However, one of the tested carbon sources, sugar beet pulp (SBP), inhibited growth of the plant pathogens, also when combined with another carbon source, while endophytic species remained unaffected.     

Evidence for a synergistic effect of post‐translational modifications and genomic composition of eEF‐1α on the adaptation of Phytophthora infestans

Genetic variation plays a fundamental role in pathogen''s adaptation to environmental stresses. Pathogens with low genetic variation tend to survive and proliferate more poorly due to their lack of genotypic/phenotypic polymorphisms in responding to fluctuating environments. Evolutionary theory hypothesizes that the adaptive disadvantage of genes with low genomic variation can be compensated for structural diversity of proteins through post‐translation modification (PTM) but this theory is rarely tested experimentally and its implication to sustainable disease management is hardly discussed. In this study, we analyzed nucleotide characteristics of eukaryotic translation elongation factor‐1α (eEF‐lα) gene from 165 Phytophthora infestans isolates and the physical and chemical properties of its derived proteins. We found a low sequence variation of eEF‐lα protein, possibly attributable to purifying selection and a lack of intra‐genic recombination rather than reduced mutation. In the only two isoforms detected by the study, the major one accounted for >95% of the pathogen collection and displayed a significantly higher fitness than the minor one. High lysine representation enhances the opportunity of the eEF‐1α protein to be methylated and the absence of disulfide bonds is consistent with the structural prediction showing that many disordered regions are existed in the protein. Methylation, structural disordering, and possibly other PTMs ensure the ability of the protein to modify its functions during biological, cellular and biochemical processes, and compensate for its adaptive disadvantage caused by sequence conservation. Our results indicate that PTMs may function synergistically with nucleotide codes to regulate the adaptive landscape of eEF‐1α, possibly as well as other housekeeping genes, in P. infestans. Compensatory evolution between pre‐ and post‐translational phase in eEF‐1α could enable pathogens quickly adapting to disease management strategies while efficiently maintaining critical roles of the protein playing in biological, cellular, and biochemical activities. Implications of these results to sustainable plant disease management are discussed.     

Hidden Behind Autophagy: The Unconventional Roles of ATG Proteins

Macroautophagy (hereafter referred to as autophagy) is an evolutionarily conserved intracellular catabolic transport route that generally allows the lysosomal degradation of cytoplasmic components, including bulk cytosol, protein aggregates, damaged or superfluous organelles and invading microbes. Target structures are sequestered by double‐membrane vesicles called autophagosomes, which are formed through the concerted action of the autophagy (ATG)‐related proteins. Until recently it was assumed that ATG proteins were exclusively involved in autophagy. A growing number of studies, however, have attributed functions to some of them that are distinct from their classical role in autophagosome biogenesis. Autophagy‐independent roles of the ATG proteins include the maintenance of cellular homeostasis and resistance to pathogens. For example, they assist and enhance the turnover of dead cells and microbes upon their phagocytic engulfment, and inhibit murine norovirus replication. Moreover, bone resorption by osteoclasts, innate immune regulation triggered by cytoplasmic DNA and the ER‐associated degradation regulation all have in common the requirement of a subset of ATG proteins. Microorganisms such as coronaviruses, Chlamydia trachomatis or Brucella abortus have even evolved ways to manipulate autophagy‐independent functions of ATG proteins in order to ensure the completion of their intracellular life cycle. Taken together these novel mechanisms add to the repertoire of functions and extend the number of cellular processes involving the ATG proteins.     

Genome sequence of the free-living aerobic spirochete Turneriella parva type strain (HT), and emendation of the species Turneriella parva

Turneriella parva Levett et al. 2005 is the only species of the genus Turneriella which was established as a result of the reclassification of Leptospira parva Hovind-Hougen et al. 1982. Together with Leptonema and Leptospira, Turneriella constitutes the family Leptospiraceae, within the order Spirochaetales. Here we describe the features of this free-living aerobic spirochete together with the complete genome sequence and annotation. This is the first complete genome sequence of a member of the genus Turneriella and the 13^th member of the family Leptospiraceae for which a complete or draft genome sequence is now available. The 4,409,302 bp long genome with its 4,169 protein-coding and 45 RNA genes is part of the Genomic Encyclopedia of Bacteria and Archaea project.     

Avrilella dinanensis gen. nov., sp. nov., a novel bacterium of the family Flavobacteriaceae isolated from human blood

Polyphasic taxonomic analysis was performed on a novel bacterium, designated UR159^T, isolated in 2016 from human blood of a septic patient hospitalized in France. Preliminary 16S rRNA gene sequence-based phylogenetic analysis indicated that strain UR159^T belonged to the family Flavobacteriaceae, forming a distinct phyletic line distantly related (<94% sequence similarity) to known species of the family. Further phenotypic, chemotaxonomic and genomic analyses were performed. Cells were non-motile, oxidase-negative, catalase-positive Gram-negative rods. It was strictly aerobic yielding yellow-pigmented colonies, and was metabolically rather inert. Major fatty acids were iso-branched fatty acids, predominantly iso-C_15:0 (55.5%) and iso-C_17:1ω9c (8.8%). Whole genome sequencing revealed a 2.3-Mbp genome encoding a total of 2262 putative genes with a genomic DNA G + C content at 37.6 mol%. The average nucleotide identity (ANI) and in silico DNA-DNA hybridization (isDDH) values between strain UR159^T and the most closely related members of the Flavobacteriaceae family were <75% and <39%, respectively, much below the established cut-offs for ANI (<95–96%) and isDDH (<70%) for species and genus delineation. Average Amino Acid Identity (AAI) percentages were also estimated and were lower than 65% (cut-off proposed for genus delineation for uncultivated prokaryotes) in all cases, except for F. marinum that was just at the limit (65.1%). Based on these findings, we propose it as a new genus and species, Avrilella dinanensis gen. nov., sp. nov. (type strain UR159^T = CIP 111616^T = DSM 105483^T).