Mutual Exclusivity of Hyaluronan and Hyaluronidase in Invasive Group A Streptococcus

A recent analysis of group A Streptococcus (GAS) invasive infections in Australia has shown a predominance of M4 GAS, a serotype recently reported to lack the antiphagocytic hyaluronic acid (HA) capsule. Here, we use molecular genetics and bioinformatics techniques to characterize 17 clinical M4 isolates associated with invasive disease in children during this recent epidemiology. All M4 isolates lacked HA capsule, and whole genome sequence analysis of two isolates revealed the complete absence of the hasABC capsule biosynthesis operon. Conversely, M4 isolates possess a functional HA-degrading hyaluronate lyase (HylA) enzyme that is rendered nonfunctional in other GAS through a point mutation. Transformation with a plasmid expressing hasABC restored partial encapsulation in wild-type (WT) M4 GAS, and full encapsulation in an isogenic M4 mutant lacking HylA. However, partial encapsulation reduced binding to human complement regulatory protein C4BP, did not enhance survival in whole human blood, and did not increase virulence of WT M4 GAS in a mouse model of systemic infection. Bioinformatics analysis found no hasABC homologs in closely related species, suggesting that this operon was a recent acquisition. These data showcase a mutually exclusive interaction of HA capsule and active HylA among strains of this leading human pathogen.     

CO2浓度升高和不同氮源对紫茎泽兰生长及光合特性的影响

大气CO2浓度升高和植物入侵是全世界面临的两大重要问题。CO2浓度升高促进植物的光合作用,但在某些植物中,这种促进作用出现在短期高浓度CO2下,而在长期高浓度CO2处理下消失（称为CO2驯化）,被认为源于高浓度CO2对光呼吸和NO-3同化的抑制。通过比较研究不同形式氮源（全氮、硝态氮）和短期（8 days）、长期（40 days）CO2浓度升高处理对入侵植物紫茎泽兰生理特征的影响,结果表明在全氮供应下,短期和长期CO2浓度升高均促进了紫茎泽兰的光合;氨态氮缺失情况下,长期CO2浓度升高促进紫茎泽兰的光合,而短期CO2浓度升高对紫茎泽兰的光合没有促进作用;缺NH＋4下,短期高浓度CO2提高了叶片叶绿素含量,长期CO2升高又使其回复到正常CO2下的较低水平。这些结果表明紫茎泽兰并不会对长期的CO2升高产生驯化,即长期CO2升高会促进紫茎泽兰的光合作用,而且这一促进作用不受土壤中缺NH＋4的影响。鉴于培养介质中缺NH＋4会导致一些植物产生“CO2驯化”,未来CO2浓度升高情况下,在缺NH＋4的土壤中,紫茎泽兰的竞争力可能会更强。     

Platelet‐rich plasma inhibits osteoblast apoptosis and actin cytoskeleton disruption induced by gingipains through upregulating integrin β1

The aim of this study was to explore the effects of platelet‐rich plasma on gingipain‐caused changes in cell morphology and apoptosis of osteoblasts. Mouse osteoblasts MC3T3‐E1 cells were treated with gingipain extracts from Porphyromonas gingivalis in the presence or absence of platelet‐rich plasma. Apoptosis was detected with terminal deoxynucleotidyl transferase‐mediated dUTP nick‐end labeling staining. F‐actin was determined by phalloidin‐fluorescent staining and observed under confocal microscopy. Western blot analysis was used to detect integrin β1, F‐actin, and G‐actin protein expressions. A knocking down approach was used to determine the role of integrin β1. The platelet‐rich plasma protected osteoblasts from gingipain‐induced apoptosis in a dose‐dependent manner, accompanied by upregulation of integrin β1. Platelet‐rich plasma reversed the loss of F‐actin integrity and decrease of F‐actin/G‐actin ratio in osteoblasts in the presence of gingipains. By contrast, the effects of platelet‐rich plasma were abrogated by knockdown of integrin β1. The platelet‐rich plasma failed to reduce cell apoptosis and reorganize the cytoskeleton after knockdown of integrin β1. In conclusion, platelet‐rich plasma inhibits gingipain‐induced osteoblast apoptosis and actin cytoskeleton disruption by upregulating integrin β1 expression.     

外来入侵植物与人工林下层植被结构的关联性

为有效防控外来入侵植物,在广西陆川县人工林区进行植被嵌套样方调查,分析群落物种组成与植被结构和环境因子之间的相关性。Mantel检验表明样方物种组成与样方植被结构的相关性(r=0.208, P=0.002)高于其与环境因子相关性(r=0.084, P=0.051)。MRPP分析表明,有入侵植物阔叶丰花草(Spermacoce alata)、假臭草(Praxelis clematidea)和胜红蓟(Ageratum conyzoides)样方的植被结构与无这3种植物样方的有极显著不同(P 0.01),而有无薇甘菊(Mikainamicrantha)样方的植被结构相似。t检验表明林冠盖度(3.0 m)与这4种植物入侵均无显著相关性(P0.073)。通过增加垂直高度1.5 m以下的植被密度,可以减少阔叶丰花草、假臭草和胜红蓟的入侵,但对抑制薇甘菊入侵无效。因此,植被结构作为过滤器,可能主要作用于植物的种子产生时间、萌发周期和生长习性;以种子方式繁殖的入侵植物能否定植相对更依赖干扰、暴露的裸地;兼具无性和有性繁殖且能攀援和匍匐生长使得薇甘菊能够在不同植被结构的群落中相对随机地定植。     

侵袭性真菌深部感染的早期实验室诊断

目的探讨假丝酵母菌甘露聚糖抗原和假丝酵母菌IgG/IgM抗体、曲霉半乳甘露聚糖抗原和烟曲霉IgG抗体在侵袭性真菌病早期临床诊断中的应用价值。方法收集已确诊侵袭性假丝酵母菌病患者18例,侵袭性烟曲霉病患者6例,单纯细菌感染患者20例,浅部真菌感染患者20例,健康体检者(正常对照组)20例,通过酶联免疫吸附法(ELISA)检测患者血清甘露聚糖和假丝酵母菌IgG/IgM抗体以及曲霉半乳甘露聚糖抗原和烟曲霉IgG抗体浓度,计算各指标的灵敏度、特异度、阳性预测值、阴性预测值和受试者工作特征(ROC)曲线下面积。结果甘露聚糖抗原和假丝酵母菌IgG/IgM抗体联合测定的敏感度为66.7%,特异度为83.3%,阴性预测值为100.0%,阳性预测值为85.7%,ROC曲线下面积为0.992(95%CI:0.974~1.000);半乳甘露聚糖抗原和烟曲霉IgG抗体联合测定的敏感度为66.7%,特异度为95.0%,阴性预测值为98.2%,阳性预测值为100.0%,ROC曲线下面积为0.978(95%CI:0.934~1.000)。结论甘露聚糖抗原和假丝酵母菌IgG/IgM抗体、半乳甘露聚糖抗原和烟曲霉IgG抗体联合检测对深部真菌感染的早期诊断具有重要意义。     

Fungal co-infection in COVID-19 patients: Should we be concerned?

Critically ill COVID-19 patients have higher pro-inflammatory (IL-1, IL-2, IL-6, tumor necrosis alpha) and anti-inflammatory (IL-4, IL-10) cytokine levels, less CD₄ interferon-gamma expression, and fewer CD₄ and CD₈ cells. This severe clinical situation increases the risk of serious fungal infections, such as invasive pulmonary aspergillosis, invasive candidiasis or Pneumocystis jirovecii pneumonia. However, few studies have investigated fungal coinfections in this population. We describe an update on published reports on fungal coinfections and our personal experience in three Spanish hospitals. We can conclude that despite the serious disease caused by SARS-CoV-2 in many patients, the scarcity of invasive mycoses is probably due to the few bronchoscopies and necropsies performed in these patients because of the high risk in aerosol generation. However, the presence of fungal markers in clinically relevant specimens, with the exception of bronchopulmonary colonization by Candida, should make it advisable to early implement antifungal therapy.     

Views on antioxidant mouthwashes as adjunct in periodontal therapy

Clinical decision is often difficult with chlorhexidine mouthwash. The use of antioxidant mouthwashes for the treatment of periodontal disease is in practise. Therefore, it is of interest to collect gleaned information on Antioxidant mouthwashes as periodontal therapy from known literature. Improvement in treatment using antioxidant mouthwashes is reported in several studies. The mouthwash with antioxidants has similar anti-gingivitis, antiplaque and antimicrobial effects as that of chlorhexidine mouthwash.     

Measuring the subgingival microbiota in periodontitis patients: Comparison of the surface layer and the underlying layers

Periodontitis is a major cause of tooth loss in adults that initially results from dental plaque. Subgingival plaque pathogenesis is affected by both community composition and plaque structures, although limited data are available concerning the latter. To bridge this knowledge gap, subgingival plaques were obtained using filter paper (the fourth layer) and curette (the first-third layers) sequentially and the phylogenetic differences between the first–third layers and the fourth layer were characterized by sequencing the V3–V4 regions of 16S rRNA. A total of 11 phyla, 148 genera, and 308 species were obtained by bioinformatic analysis, and no significant differences between the operational taxonomic unit numbers were observed for these groups. In both groups, the most abundant species were Porphyromonas gingivalis and Fusobacterium nucleatum. Actinomyces naeslundii, Streptococcus intermedius, and Prevotella intermedia possessed relatively high proportions in the first–third layers; while in the fourth layer, both traditional pathogens (Treponema denticola and Campylobacter rectus) and novel pathobionts (Eubacterium saphenum, Filifactor alocis, Treponema sp. HOT238) were prominent. Network analysis showed that either of them exhibited a scale-free property and was constructed by two negatively correlated components (the pathogen component and the nonpathogen component), while the synergy in the nonpathogen component was lower in the first–third layers than that in the fourth layer. After merging these two parts into a whole plaque group, the negative/positive correlation ratio increased. With potential connections, the first–third layers and the fourth layer showed characteristic key nodes in bacterial networks.     

Pressure Cycling Technology Assisted Mass Spectrometric Quantification of Gingival Tissue Reveals Proteome Dynamics during the Initiation and Progression of Inflammatory Periodontal Disease

Understanding the progression of periodontal tissue destruction is at the forefront of periodontal research. The authors aimed to capture the dynamics of gingival tissue proteome during the initiation and progression of experimental (ligature‐induced) periodontitis in mice. Pressure cycling technology (PCT), a recently developed platform that uses ultra‐high pressure to disrupt tissues, is utilized to achieve efficient and reproducible protein extraction from ultra‐small amounts of gingival tissues in combination with liquid chromatography‐tandem mass spectrometry (MS). The MS data are processed using Progenesis QI and the regulated proteins are subjected to METACORE, STRING, and WebGestalt for functional enrichment analysis. A total of 1614 proteins with ≥2 peptides are quantified with an estimated protein false discovery rate of 0.06%. Unsupervised clustering analysis shows that the gingival tissue protein abundance is mainly dependent on the periodontitis progression stage. Gene ontology enrichment analysis reveals an overrepresentation in innate immune regulation (e.g., neutrophil‐mediated immunity and antimicrobial peptides), signal transduction (e.g., integrin signaling), and homeostasis processes (e.g., platelet activation and aggregation). In conclusion, a PCT‐assisted label‐free quantitative proteomics workflow that allowed cataloging the deepest gingival tissue proteome on a rapid timescale and provided novel mechanistic insights into host perturbation during periodontitis progression is applied.     

对新型冠状病毒肺炎继发真菌感染的思考

新型冠状病毒肺炎(coronavirus disease 2019,COVID-19)疫情十分严重,给民众的健康带来了巨大威胁。COVID-19患者可能继发侵袭性真菌感染,会严重威胁患者的生命,因此,在诊治策略上应该给予重视。除了加强高危患者中病原真菌的常规检查外,还应加大力度支持和扶持病原真菌先进检测技术的研发;此外,还应重点支持针对医疗单位、公共场所和家庭等常温环境以及体表和器物表面灭活病毒、细菌、真菌等病原体的新型技术和方法的研发。最终为国家战胜COVID-19和继发感染疫情提供新措施和新策略。