Integrated Metabolomics and Transcriptomics Analyses Reveal Metabolic Landscape in Neuronal Cells during JEV Infection

Virologica Sinica - Japanese encephalitis virus (JEV) is a leading cause of viral encephalitis in endemic regions of Asia. The neurotropism of JEV and its high-efficiency replication in neurons are...     

Autophagy Enhances Bacterial Clearance during P. aeruginosa Lung Infection

Pseudomonas aeruginosa is an opportunistic bacterial pathogen which is the leading cause of morbidity and mortality among cystic fibrosis patients. Although P. aeruginosa is primarily considered an extacellular pathogen, recent reports have demonstrated that throughout the course of infection the bacterium acquires the ability to enter and reside within host cells. Normally intracellular pathogens are cleared through a process called autophagy which sequesters and degrades portions of the cytosol, including invading bacteria. However the role of autophagy in host defense against P. aeruginosa in vivo remains unknown. Understanding the role of autophagy during P. aeruginosa infection is of particular importance as mutations leading to cystic fibrosis have recently been shown to cause a blockade in the autophagy pathway, which could increase susceptibility to infection. Here we demonstrate that P. aeruginosa induces autophagy in mast cells, which have been recognized as sentinels in the host defense against bacterial infection. We further demonstrate that inhibition of autophagy through pharmacological means or protein knockdown inhibits clearance of intracellular P. aeruginosa in vitro, while pharmacologic induction of autophagy significantly increased bacterial clearance. Finally we find that pharmacological manipulation of autophagy in vivo effectively regulates bacterial clearance of P. aeruginosa from the lung. Together our results demonstrate that autophagy is required for an effective immune response against P. aeruginosa infection in vivo, and suggest that pharmacological interventions targeting the autophagy pathway could have considerable therapeutic potential in the treatment of P. aeruginosa lung infection.     

Defining Epidermal Basal Cell States during Skin Homeostasis and Wound Healing Using Single-Cell Transcriptomics

1. Download : Download high-res image (254KB)
2. Download : Download full-size image

     

Bacterial Metabolism of Mevalonic Acid

Soluble cell-free extracts of actinomycete S4 grown on media containing mevalonate catalyze acetoacetate formation from mevalonate, mevaldate, and β-hydroxy-β-methylglutaryl-coenzyme A (CoA). Conversion of mevalonate to acetoacetate involves formation of free β-hydroxy-β-methylglutaryl-CoA, but not free mevaldate. The reaction favors mevalonate oxidation, and nicotinamide adenine dinucleotide, rather than nicotinamide adenine dinucleotide phosphate, acts as oxidant.     

Bacterial Metabolism of 2,6-Xylenol

Strain DM1, a Mycobacterium sp. that utilizes 2,6-xylenol, 2,3,6-trimethylphenol, and o-cresol as sources of carbon and energy, was isolated. Intact cells of Mycobacterium strain DM1 grown with 2,6-xylenol cooxidized 2,4,6-trimethylphenol to 2,4,6-trimethylresorcinol. 4-Chloro-3,5-dimethylphenol prevents 2,6-xylenol from being totally degraded; it was quantitatively converted to 2,6-dimethylhydroquinone by resting cells. 2,6-Dimethylhydroquinone, citraconate, and an unidentified metabolite were detected as products of 2,6-xylenol oxidation in cells that were partially inactivated by EDTA. Under oxygen limitation, 2,6-dimethylhy-droquinone, citraconate, and an unidentified metabolite were released during 2,6-xylenol turnover by resting cells. Cell extracts of 2,6-xylenol-grown cells contained a 2,6-dimethylhydroquinone-converting enzyme. When supplemented with NADH, cell extracts catalyzed the reduction of 2,6-dimethyl-3-hydroxyquinone to 2,6-dimethyl-3-hydroxyhydroquinone. Since a citraconase was also demonstrated in cell extracts, a new metabolic pathway with 2,6-dimethyl-3-hydroxyhydroquinone as the ring fission substrate is proposed.     

Heterogeneity in Bacterial Specialized Metabolism

Specialized metabolites (SMs) like typical antibiotics, signaling molecules or other bioactive compounds of bacterial origin (sometimes also used in human therapy) are often complex natural products that are costly for the cell to make. Several bacterial taxa are known to produce multiple SM classes in parallel and therefore a division of labor within a clonal population of bacteria might be beneficial. In this review, examples of SM of gram-negative and gram-positive bacterial taxa that are produced by different cell types are presented, and the possibility that such a heterogeneity is more widespread in SM biosynthesis is discussed. In addition, tools to study SM production at the single cell level are presented.     

High-Resolution Underwater Mapping Using Side-Scan Sonar

The goal of this study is to generate high-resolution sea floor maps using a Side-Scan Sonar(SSS). This is achieved by explicitly taking into account the SSS operation as follows. First, the raw sensor data is corrected by means of a physics-based SSS model. Second, the data is projected to the sea-floor. The errors involved in this projection are thoroughfully analysed. Third, a probabilistic SSS model is defined and used to estimate the probability of each sea-floor region to be observed. This probabilistic information is then used to weight the contribution of each SSS measurement to the map. Because of these models, arbitrary map resolutions can be achieved, even beyond the sensor resolution. Finally, a geometric map building method is presented and combined with the probabilistic approach. The resulting map is composed of two layers. The echo intensity layer holds the most likely echo intensities at each point in the sea-floor. The probabilistic layer contains information about how confident can the user or the higher control layers be about the echo intensity layer data. Experimental results have been conducted in a large subsea region.     

子宫内膜与细菌L型感染

本文报道41例宫腔刮出物的病原微生物培养,其中子宫异常出血和早孕胎盘组织各15例,月经紊乱6例和不孕症5例。19例细菌培养为阳性(46.35),19例中之7例为细菌兼有 L 型感染,8例为单纯 L 型感染,3例为解脲脲原体和1例 G~ 厌氧球菌感染。L 型菌落呈典型“油煎蛋”样或颗粒样。子宫内膜和胎盘组织切片的细菌学检查发现大量 L 型巨形体、圆球体和长丝体。作者认为在妇产科范围内,细菌 L 型和解脲脲原体感染并非少见,可能是子宫异常出血、月经紊乱、不孕症和流产的重要原因之一。L 型的形成与抗生素治疗及体内因素的诱导有关。在治疗 L 型感染过程中,药物的选择很重要。     

High-Resolution Melt Analysis for Rapid Comparison of Bacterial Community Compositions

In the study of bacterial community composition, 16S rRNA gene amplicon sequencing is today among the preferred methods of analysis. The cost of nucleotide sequence analysis, including requisite computational and bioinformatic steps, however, takes up a large part of many research budgets. High-resolution melt (HRM) analysis is the study of the melt behavior of specific PCR products. Here we describe a novel high-throughput approach in which we used HRM analysis targeting the 16S rRNA gene to rapidly screen multiple complex samples for differences in bacterial community composition. We hypothesized that HRM analysis of amplified 16S rRNA genes from a soil ecosystem could be used as a screening tool to identify changes in bacterial community structure. This hypothesis was tested using a soil microcosm setup exposed to a total of six treatments representing different combinations of pesticide and fertilization treatments. The HRM analysis identified a shift in the bacterial community composition in two of the treatments, both including the soil fumigant Basamid GR. These results were confirmed with both denaturing gradient gel electrophoresis (DGGE) analysis and 454-based 16S rRNA gene amplicon sequencing. HRM analysis was shown to be a fast, high-throughput technique that can serve as an effective alternative to gel-based screening methods to monitor microbial community composition.     

轮状病毒感染患者外周血单个核细胞的转录组学分析

轮状病毒(Rotavirus,RV)是引起急性肠胃炎的主要病原体,分析RV感染患者的人外周血单个核细胞(Peripheral blood mononuclear cell,PBMC)中差异表达基因(Differentially expressed genes,DEGs)有利于探讨人PBMC在清除RV中的作用。为此,本研究采集2019年2月-2019年6月长春儿童医院中RV感染患者和健康儿童血液,分离PBMC,通过转录组测序(RNA sequencing,RNA-seq)技术比较RV感染患者与健康儿童之间的RNA表达图谱,借助基因本体论(Gene Ontology,GO)数据库功能富集分析、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)、Reactome通路富集分析DEGs,使用实时荧光定量PCR(Real-time quantitative PCR,qPCR)技术进行验证。结果显示,与健康对照组相比,RV感染轻症患者PBMC中有1619个DEGs;重症患者PBMC中有2816个DEGs,主要与干扰素(Interferon,IFN)反应、中性粒细胞、溶酶体、核小体、染色质等相关。qPCR验证轻症患者干扰素刺激基因(IFN-stimulated genes,ISGs)15表达上调,白介素(Interleukin,IL)1β表达下调;重症患者IL15、ISG15表达上调,IL1β表达下调,与转录组结果相一致。本研究提示,RV感染可能激活人I型和II型IFNs反应抵御病毒感染,但也会抑制溶酶体相关基因,对细胞自噬过程产生影响。