Genetic targeting of Card19 is linked to disrupted NINJ1 expression,impaired cell lysis,and increased susceptibility to Yersinia infection

Cell death plays a critical role in inflammatory responses. During pyroptosis, inflammatory caspases cleave Gasdermin D (GSDMD) to release an N-terminal fragment that generates plasma membrane pores that mediate cell lysis and IL-1 cytokine release. Terminal cell lysis and IL-1β release following caspase activation can be uncoupled in certain cell types or in response to particular stimuli, a state termed hyperactivation. However, the factors and mechanisms that regulate terminal cell lysis downstream of GSDMD cleavage remain poorly understood. In the course of studies to define regulation of pyroptosis during Yersinia infection, we identified a line of Card19-deficient mice (Card19^lxcn) whose macrophages were protected from cell lysis and showed reduced apoptosis and pyroptosis, yet had wild-type levels of caspase activation, IL-1 secretion, and GSDMD cleavage. Unexpectedly, CARD19, a mitochondrial CARD-containing protein, was not directly responsible for this, as an independently-generated CRISPR/Cas9 Card19 knockout mouse line (Card19^Null) showed no defect in macrophage cell lysis. Notably, Card19 is located on chromosome 13, immediately adjacent to Ninj1, which was recently found to regulate cell lysis downstream of GSDMD activation. RNA-seq and western blotting revealed that Card19^lxcn BMDMs have significantly reduced NINJ1 expression, and reconstitution of Ninj1 in Card19^lxcn immortalized BMDMs restored their ability to undergo cell lysis in response to caspase-dependent cell death stimuli. Card19^lxcn mice exhibited increased susceptibility to Yersinia infection, whereas independently-generated Card19^Null mice did not, demonstrating that cell lysis itself plays a key role in protection against bacterial infection, and that the increased infection susceptibility of Card19^lxcn mice is attributable to loss of NINJ1. Our findings identify genetic targeting of Card19 being responsible for off-target effects on the adjacent gene Ninj1, disrupting the ability of macrophages to undergo plasma membrane rupture downstream of gasdermin cleavage and impacting host survival and bacterial control during Yersinia infection.     

Mass spectrometry-based proteomic approaches to study pathogenic bacteria-host interactions

Elucidation of molecular mechanisms underlying hostpathogen interactions is important for control and treatment of infectious diseases worldwide. Within the last decade, mass spectrometry (MS)-based proteomics has become a powerful and effective approach to better understand complex and dynamic host-pathogen interactions at the protein level. Herein we will review the recent progress in proteomic analyses towards bacterial infection of their mammalian host with a particular focus on enteric pathogens. Large-scale studies of dynamic proteomic alterations during infection will be discussed from the perspective of both pathogenic bacteria and host cells.     

土壤原生动物对川滇高山栎恢复时间的响应及生长季动态

青藏高原东缘生态环境脆弱, 森林频繁遭到砍伐, 生物多样性受到严重威胁, 森林砍伐后的生态恢复成为研究热点。原生动物在生态恢复中作为指示生物起着重要作用。本文就未砍伐、砍伐后不同恢复时期(20年、10年和1年)生的川滇高山栎(Quercus aquifolioides)林的土壤理化性质和原生动物在生长季的变化进行比较研究, 以探讨生长季不同月份、不同恢复期的原生动物数量变化规律, 分析土壤理化性质与其相关性。结果表明: (1)恢复10年和1年的林地的鞭毛虫数量(193个/g干土, 164个/g干土)显著高于原始林地(22个/g干土), 肉足虫在恢复1年的林地中数量最多(600个/g干土), 纤毛虫数量则随次生演替进程逐渐增多。(2)在生长季不同月份原生动物的数量呈先增加后减少的趋势。鞭毛虫和肉足虫的峰值分别出现在7月和8月, 而纤毛虫的数量在7、8、9月明显高于6月。(3)原生动物数量与土壤理化性质密切相关。鞭毛虫数量与pH值呈显著正相关(P = 0.019), 纤毛虫数量与铵态氮(P = 0.002)和碳氮比呈显著正相关(P = 0.022), 肉足虫数量与硝态氮(P = 0.008)和碳氮比(P = 0.016)呈显著负相关。结果显示, 三种原生动物数量在生长季不同月份表现出较大的波动性, 其数量变化受土壤理化性质等多种因素调控。纤毛虫数量对川滇高山栎林砍伐恢复有正响应, 而鞭毛虫、肉足虫数量有负响应。     

TNF‐α induces up‐regulation of MicroRNA‐27a via the P38 signalling pathway,which inhibits intervertebral disc degeneration by targeting FSTL1

The mechanism of intervertebral disc degeneration is still unclear, and there are no effective therapeutic strategies for treating this condition. miRNAs are naturally occurring macromolecules in the human body and have many biological functions. Therefore, we hope to elucidate whether miRNAs are associated with intervertebral disc degeneration and the underlying mechanisms involved. In our study, differentially expressed miRNAs were predicted by the GEO database and then confirmed by qPCR and in situ hybridization. Apoptosis of nucleus pulposus cells was detected by flow cytometry and Bcl2, Bax and caspase 3. Deposition of extracellular matrix was assessed by Alcian blue staining, and the expression of COX2 and MMP13 was detected by immunofluorescence, Western blot and qPCR. Moreover, qPCR was used to detect the expression of miR27a and its precursors. The results showed that miR27a was rarely expressed in healthy intervertebral discs but showed increased expression in degenerated intervertebral discs. Ectopic miR27a expression inhibited apoptosis, suppressed the inflammatory response and attenuated the catabolism of the extracellular matrix by targeting FSTL1. Furthermore, it seems that the expression of miR27a was up‐regulated by TNF‐α via the P38 signalling pathway. So we conclude that TNF‐α and FSTL1 engage in a positive feedback loop to promote intervertebral disc degeneration. At the same time, miR27a is up‐regulated by TNF‐α via the P38 signalling pathway, which ameliorates inflammation, apoptosis and matrix degradation by targeting FSTL1. Thus, this negative feedback mechanism might contribute to the maintenance of a low degeneration load and would be beneficial to maintain a persistent chronic disc degeneration.     

The relative importance of architecture and resource competition in allocation to pollen and ovule number within inflorescences of Hosta ventricosa varies with the resource pools

Background and Aims

Allocation of resources to floral traits often declines distally within inflorescences in flowering plants. Architecture and resource competition have been proposed as underlying mechanisms. The aim of the present study is to assess the relative importance of resource competition and architectural effects in pollen and ovule production on racemes of Hosta ventricosa, an apomictic perennial herb.

Methods

Combinations of two defoliation treatments (intact and defoliated) and two fruit-set treatments (no-fruit and fruit) were created, and the roles of architecture and resource competition at each resource level were assessed.

Key Results

Pollen and ovule number per flower increased after defoliation, but pollen to ovule ratio per flower did not change. Pollen, ovules and the pollen to ovule ratio per flower declined distally on racemes at each resource level. In the intact treatment, fruit development of early flowers did not affect either pollen or ovule number of late flowers. In the defoliated treatment, fruit development of early flowers reduced both pollen and ovule numbers of late flowers due to over-compensation caused by defoliation. Late flowers on defoliated fruit racemes produced less pollen than intact fruit racemes but the same number of ovules; therefore, the reduction in pollen number was not caused by over-compensation. In addition, the fruit-set rate of early flowers during flowering was higher in intact racemes than in defoliated racemes.

Conclusions

In flowering plants, the relative importance of architecture and resource competition in allocation to pollen and ovules may vary with the resource pools or the overall resource availability of maternal plants.     

岷江上游半干旱河谷区3种林型土壤氮素的比较

比较研究了岷江上游半干旱河谷区辐射松人工林、油松人工林与邻近灌丛0-20cm、20-40cm、40-60cm土层土壤氮素和氮循环过程相关酶的特征,包括土壤有机碳、全氮、碳氮比、硝态氮、铵态氮、无机氮、微生物量氮含量及蛋白酶、脲酶、硝酸盐还原酶活性。结果表明,辐射松林和油松林各土层土壤有机碳含量、碳氮比和硝酸盐还原酶活性无显著差异,油松林土壤无机氮含量和脲酶活性显著高于辐射松林土壤,而辐射松林土壤微生物量氮含量是3种林型土壤中最高的,灌丛0-20cm土层土壤有机碳、全氮含量最高。此外,有机碳、全氮含量、脲酶活性随土层深度增加而降低;而硝酸盐还原酶活性却随土层深度的增加而增强;同时,各土层间蛋白酶活性差异较小。因此,植被类型对土壤氮素转化有一定影响,而从目前的土壤氮素状况来看,油松林土壤中植物可直接吸收利用的氮素高于辐射松林和灌丛;辐射松林土壤微生物固持的氮素含量最高。区域3种植被类型土壤氮素状况还受到半干旱气候因素的强烈影响。     

生姜水浸液对生姜幼苗根际土壤酶活性、微生物群落结构及土壤养分的影响

本文以生姜为材料,研究生姜不同部位（根茎、茎和叶）、不同浓度（10、20、40和80 g L-1）的水浸液对生姜幼苗根际土的微生物数量、土壤酶活性及土壤养分含量的影响,并通过HPLC定量分析了生姜各部位水浸液中所含酚酸类（香草酸、丁香酸、对羟基苯甲酸、香豆酸和阿魏酸）、香豆素类（伞花内脂和7-甲氧基香豆素）化合物的含量。结果表明：三种生姜水浸液对所测六种土壤酶活性均产生了不同程度的影响,其中影响最大的是酸性磷酸酶和蔗糖酶,在10 g L-1 时就达到了显著性差异水平,并且所有酶均有随着水浸液浓度的增加而呈增大的趋势;相同部位的水浸液随着浓度的增加,细菌和真菌的数量呈增加趋势,而放线菌的数量呈减少趋势;三种生姜水浸液均随着浓度的增加降低了土壤中有机质的含量,加剧了土壤中硝态氮含量的积累,根茎水浸液对土壤有效磷、速效钾和铵态氮均显示出低浓度提高其含量而高浓度降低其含量的趋势,而茎和叶水浸液则随着浓度的增加均降低了其含量。生姜水浸液中主要化感成分包括：根茎水浸液主要是丁香酸和伞花内脂;茎水浸液主要是阿魏酸,且其含量最高为73.4 ug/g;叶水浸液除了阿魏酸,其他六种物质均被检测出,但含量较高的主要有丁香酸、伞花内脂和香豆酸。     

花椒叶浸提液对土壤微生物数量和土壤酶活性的影响

通过用花椒叶浸提液浇灌盆栽花椒幼苗,研究浸提液对土壤酶和土壤微生物的影响.结果表明, 花椒叶浸提液使根际土中细菌、真菌和放线菌数量以及微生物总数均有不同程度的减少,根际土中真菌和放线菌的数量变化呈降-升-降-升的趋势.20、60和80 g·L^-1浓度的叶浸提液使非根际土中细菌的数量显著增加21.59%、107.55%和8.62%,而40 g·L-1浓度的叶浸提液则使非根际土中细菌数量显著降低22.56%.叶浸提液使根际土蛋白酶、蔗糖酶和酸性磷酸酶活性明显低于非根际土相应的酶活性,而过氧化氢酶和多酚氧化酶活性则显著升高.土壤的蛋白酶活性与蔗糖酶活性呈显著正相关,与土壤放线菌数量呈显著负相关;多酚氧化酶活性与蔗糖酶活性呈显著负相关,与细菌、真菌、放线菌以及微生物总数呈显著正相关;放线菌只与蛋白酶、多酚氧化酶、蔗糖酶3种酶活性及真菌呈显著相关,与过氧化氢酶、酸性磷酸酶以及细菌和微生物总数的相关性均不显著.     

Pull-push neuromodulation of LTP and LTD enables bidirectional experience-induced synaptic scaling in visual cortex

Neuromodulatory input, acting on G protein-coupled receptors, is essential for the induction of experience-dependent cortical plasticity. Here we report that G-coupled receptors in layer II/III of visual cortex control the polarity of synaptic plasticity through a pull-push regulation of LTP and LTD. In slices, receptors coupled to Gs promote LTP while suppressing LTD; conversely, receptors coupled to Gq11 promote LTD and suppress LTP. In vivo, the selective stimulation of Gs- or Gq11-coupled receptors brings the cortex into LTP-only or LTD-only states, which allows the potentiation or depression of targeted synapses with visual stimulation. The pull-push regulation of LTP/LTD occurs via direct control of the synaptic plasticity machinery and it is independent of changes in NMDAR activation or neuronal excitability. We propose these simple rules governing the pull-push control of LTP/LTD form a general metaplasticity mechanism that may contribute to neuromodulation of plasticity in other cortical circuits.