TLR-2/TLR-4 TREM-1 Signaling Pathway Is Dispensable in Inflammatory Myeloid Cells during Sterile Kidney Injury

Inflammatory macrophages are abundant in kidney disease, stimulating repair, or driving chronic inflammation and fibrosis. Damage associated molecules (DAMPs), released from injured cells engage pattern recognition receptors (PRRs) on macrophages, contributing to activation. Understanding mechanisms of macrophage activation during kidney injury may lead to strategies to alleviate chronic disease. We identified Triggering-Receptor-in-Myeloid-cells (TREM)-1, a regulator of TLR signaling, as highly upregulated in kidney inflammatory macrophages and tested the roles of these receptors in macrophage activation and kidney disease. Kidney DAMPs activated macrophages in vitro, independently of TREM-1, but partially dependent on TLR-2/−4, MyD88. In two models of progressive interstitial kidney disease, TREM-1 blockade had no impact on disease or macrophage activation in vivo, but TLR-2/−4, or MyD88 deficiency was anti-inflammatory and anti-fibrotic. When MyD88 was mutated only in the myeloid lineage, however, there was no bearing on macrophage activation or disease progression. Instead, TLR-2/−4 or MyD88 deficiency reduced activation of mesenchyme lineage cells resulting in reduced inflammation and fibrosis, indicating that these pathways play dominant roles in activation of myofibroblasts but not macrophages. To conclude, TREM-1, TLR2/4 and MyD88 signaling pathways are redundant in myeloid cell activation in kidney injury, but the latter appear to regulate activation of mesenchymal cells.     

Role of serum Metadherin mRNA expression in the diagnosis and prediction of survival in patients with colorectal cancer

Molecular Biology Reports - Early diagnosis and treatment of colorectal cancer (CRC) are important for improving patients’ survival. Metadherin is an oncogene that plays a pivotal role in...     

Cell lines, Md108 and Md66, from the hemocytes of Malacosoma disstria (Lepidoptera) display aspects of plasma-free innate non-self activities

The innate non-self response systems of the deciduous tree pest, the forest tent caterpillar, Malacosoma disstria has been documented by us in terms of in vitro and in vivo reactions towards the Gram-positive nonpathogenic bacterium, Bacillus subtilis and Gram-negative pathogenic microbe, Xenorhabdus nematophila and their respective surface antigens, lipopoteichoic acids (LTA) and lipopolysaccharides (LPS). These studies, often conducted in whole and diluted hemolymph, preclude examination of plasma-free cellular (hemocyte) responses. Plasma-free hemocytes as primary cultures are difficult to obtain. The floating cell line Md66 and attached cell line Md108 from M. disstria hemocytes were examined as a model for plasma-free M. disstria hemocyte non-self responses. Herein, it was established that although both lines differed from each other and from the primary hemocyte cultures of M. disstria in growth parameters, cell composition and sizes both cell lines displayed granular cell-like (GL) cells and plasmatocyte-like (PL) cells according to morphological criteria and to some extent antigenic similarities based on labeling with anti-Chrysodeixis includens hemocyte monoclonal antibodies. Hemocyte-specific neuroglian-like protein was detected on cells of both cell lines and in the primary hemocyte cultures albeit with staining patterns differing according to culture and cell types, confluency levels and cell–cell adhesion. Both cell lines bound B. subtilis and X. nematophila, the reaction extent varying with the cell line and its cell types. LPS damaged both cell types in the two cell lines whereas LTA enhanced the adhesion of Md66 GL cells to flask surfaces followed by PL cell adhesion. PL cells of both lines, like the primary cultures, phagocytosed FITC-labeled B. subtilis; only Md108 GL cells phagocytosed B. subtilis. In either case phagocytosis was always less in frequency and intensity than the primary cultures. Proteins released from the cell lines differed in pattern and magnitude but contained bacterial binding proteins that enhanced differential bacterial adhesion to both cell types in both cell lines: the GL cells both cultures, and those of granular cells in primary cultures, were more involved than the primary plasmatocytes and PL cells. Only Md66 cells possessed lysozyme and both cell types of both lines contained phenoloxidase. Neither enzyme type was released during early phase reaction with the bacteria. LPS inhibited phenoloxidase activity. The similarities and differences between the lines and primary cultures make Md66 and Md108 useful for the systematic examination of plasma-free cellular non-self reactions.     

WISE 2005: chronic bed rest impairs microcirculatory endothelium in women

Sedentary behavior has deleterious effects on the cardiovascular system, including reduced endothelial functions. A 2-mo bed rest study in healthy women [women international space simulation for exploration (WISE) 2005 program] presented a unique opportunity to analyze the specific effects of prolonged inactivity without other vascular risk factors on the endothelium. We investigated endothelial properties before and after 56 days of bed rest in 8 subjects who performed no exercise (control group: No-EX) and in 8 subjects who regularly performed treadmill exercise in a lower body negative pressure chamber as well as resistance exercise (countermeasure group, EX). A functional evaluation of the microcirculation in the skin was assessed with laser Doppler. We studied endothelium-dependent and -independent vasodilation using iontophoresis of acetylcholine and sodium nitroprusside, respectively. We also measured circulating endothelial cells (CECs), an index of endothelial damage. In the No-EX group, endothelium-dependent vasodilation was significantly reduced (35.4 +/- 4.8% vs. 24.1 +/- 3.8%, P < 0.05) by bed rest with a significant increase in the number of CECs (3.6 +/- 1.4 vs. 10.6 +/- 2.7 ml(-1), P < 0.05). In the EX group, endothelium-dependent vasodilation and number of CECs were preserved. Our study shows that in humans prolonged bed rest causes impairment of endothelium-dependent function at the microcirculatory level, along with an increase in circulating endothelial cells. Microcirculatory endothelial dysfunction might participate in cardiovascular deconditioning, as well as in several bed rest-induced pathologies. We therefore conclude that the endothelium should be a target for countermeasures during periods of prolonged deconditioning.     

黑线毛足鼠年龄和种群密度与肝毛细线虫感染率的关系

2004—2005年,在内蒙古锡林郭勒盟阿巴嘎旗和东乌珠穆沁旗研究了肝毛细线虫病对黑线毛足鼠种群感染情况,分析肝毛细线虫对黑线毛足鼠的感染率与鼠体年龄以及种群密度的关系。结果表明,黑线毛足鼠达到一定的年龄(或体质量)才可感染肝毛细线虫病,最低感染个体体质量为14.6g。肝毛细线虫对低龄鼠的感染检出率比较低,而对成体鼠感染检出率较高,其感染率和感染度均随着个体年龄的增长而增高(感染率与年龄:r=0.97,P<0.05;感染度与年龄:r=0.93,P<0.05)。此外,黑线毛足鼠体质量与肝毛细线虫感染率和感染度也存在显著的相关关系(感染率与体质量:r=0.99,P<0.05;感染度与体质量:r=0.95,P<0.05),黑线毛足鼠的种群密度则对肝毛细线虫的感染率(r=0.27,P>0.05)和平均感染度(r=0.41,P>0.05)没有明显的影响,其感染率可能与地区不同有关。     

用19个生化基因位点研究普通级Z:ZCLA长爪沙鼠的遗传多样性

目的调查Z:ZCLA长爪沙鼠原种群普通级长爪沙鼠的遗传多样性。方法用本实验室自行建立的长爪沙鼠19个生化基因位点的乙酸纤维素膜电泳技术并结合基本群体遗传学指标研究了普通级Z:ZCLA长爪沙鼠100个家系的遗传多态性。结果Z:ZCLA长爪沙鼠在Es-1、Car-2、Hbb、Gpi-1、Cs-1、Ce-2I、dh-l、Mod-1呈单态,在Es-2、Es-3、Es-4、Es-6、Es-8、Es-9、Es-10、pd-1、gm-1、Trf、Akp-1个位点上呈现多态性,等位基因从2~4个不等,平均等位基因数3.0,平均杂合度0.512,平均多态信息量0.455。结论提示目前本群遗传多样性水平处于中度多态。     

荒漠草原人工柠条林对冠下草本植物群落的保育作用

以宁夏盐池县荒漠草原人工柠条（Caragana intermedia）林为研究对象,分别选取柠条林冠下东侧（SE）,冠下西侧（SW）及带间（Gap）为研究样地,从群落水平探讨柠条对冠下草本植物群落结构、物种多样性及功能群分布的影响。结果表明：（1）在3种微生境中均鉴定出12种植物,SW以蒙古冰草为优势种,SE以蒙古冰草和中亚白草为优势种,Gap则以蒙古冰草和牛枝子为优势种。（2）与Gap相比,SW和SE植物群落的平均高度分别增加了41.06%和81.75%,地上生物量分别增加了40.88%和38.73%。SW和SE中,禾本科植物地上生物量分别占地上总生物量的67.10%和58.40%,显著高于Gap （P<0.05）。（3）柠条冠层效应使得草本植物的物种丰富度指数增加,但Shannon-Winner指数、Simpson指数和Pielou指数显著差异（P>0.05）,变化范围分别为1.620-1.756、0.701-0.730和0.775-0.878。（4）冗余分析表明：土壤温度、空气相对湿度及土壤有机碳是影响草本植物物种多样性及生物量的主要因子,解释量分别为42.70%,11.70%和8.80%。研究表明,柠条对冠下草本植物群落尤其是禾本科植物具有一定的保育作用,该效应的产生主要是由于柠条冠下微气候及土壤环境因子的改善为草本植物的生长发育提供了有利条件。柠条对草本植物的保育作用对荒漠草原生态环境的保护与恢复具有重要意义。     

丹参内生拮抗细菌DS-R5对丹参根际和根表土壤细菌群落结构的影响CSCD

目的 探究生防细菌DS-R5施入丹参植株后根际和根表土壤细菌群落组成及多样性变化。方法 向丹参植株根部施入生防细菌DS-R5,以未施用细菌为对照组,分别采集根际和根表土壤样品提取总DNA,扩增样品总DNA的V3-V4区,采用Illumina MiSeq测序平台对PCR扩增产物进行双端测序分析,利用生物信息学分析解析丹参植株根际土壤和根表土壤细菌群落结构组成及多样性。结果 菌株DS-R5处理后增加了根际土壤细菌群落的多样性和丰度,降低了根表土壤细菌群落的多样性和丰度;高通量测序得到的根际和根表土壤的有效序列数量和OTU数量相比对照组均有所下降,根际土壤处理样品中微生物种类最丰富,根表土壤处理样品中微生物种类最少,根际土壤处理样品与根际土壤对照物种种类更接近;在门水平上,根际土壤处理样品相比对照变形菌门丰度下降,酸杆菌门丰度升高,根表土壤处理样品相比对照变形菌门和酸杆菌门丰度均升高,放线菌门丰度降低;在属水平上,根际土壤处理样品中鞘氨醇单胞菌属、芽胞杆菌属、慢生根瘤菌属相比根际土壤对照占比均有升高,根表土壤处理样品相比对照黄杆菌属和伯克菌属丰度下降,而土壤中的优势菌属根瘤菌属和芽胞杆菌属丰度升高。结论 丹参植株施用生防细菌DS-R5后,改变了根际土壤和根表土壤中微生物群落结构和多样性。     

Genome sequencing and comparative analysis of three Chlamydia pecorum strains associated with different pathogenic outcomes

Background

Chlamydia pecorum is the causative agent of a number of acute diseases, but most often causes persistent, subclinical infection in ruminants, swine and birds. In this study, the genome sequences of three C. pecorum strains isolated from the faeces of a sheep with inapparent enteric infection (strain W73), from the synovial fluid of a sheep with polyarthritis (strain P787) and from a cervical swab taken from a cow with metritis (strain PV3056/3) were determined using Illumina/Solexa and Roche 454 genome sequencing.

Results

Gene order and synteny was almost identical between C. pecorum strains and C. psittaci. Differences between C. pecorum and other chlamydiae occurred at a number of loci, including the plasticity zone, which contained a MAC/perforin domain protein, two copies of a >3400 amino acid putative cytotoxin gene and four (PV3056/3) or five (P787 and W73) genes encoding phospholipase D. Chlamydia pecorum contains an almost intact tryptophan biosynthesis operon encoding trpABCDFR and has the ability to sequester kynurenine from its host, however it lacks the genes folA, folKP and folB required for folate metabolism found in other chlamydiae. A total of 15 polymorphic membrane proteins were identified, belonging to six pmp families. Strains possess an intact type III secretion system composed of 18 structural genes and accessory proteins, however a number of putative inc effector proteins widely distributed in chlamydiae are absent from C. pecorum. Two genes encoding the hypothetical protein ORF663 and IncA contain variable numbers of repeat sequences that could be associated with persistence of infection.

Conclusions

Genome sequencing of three C. pecorum strains, originating from animals with different disease manifestations, has identified differences in ORF663 and pseudogene content between strains and has identified genes and metabolic traits that may influence intracellular survival, pathogenicity and evasion of the host immune system.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-23) contains supplementary material, which is available to authorized users.     

Association of Genetic Loci with Sleep Apnea in European Americans and African-Americans: The Candidate Gene Association Resource (CARe)

Although obstructive sleep apnea (OSA) is known to have a strong familial basis, no genetic polymorphisms influencing apnea risk have been identified in cross-cohort analyses. We utilized the National Heart, Lung, and Blood Institute (NHLBI) Candidate Gene Association Resource (CARe) to identify sleep apnea susceptibility loci. Using a panel of 46,449 polymorphisms from roughly 2,100 candidate genes on a customized Illumina iSelect chip, we tested for association with the apnea hypopnea index (AHI) as well as moderate to severe OSA (AHI≥15) in 3,551 participants of the Cleveland Family Study and two cohorts participating in the Sleep Heart Health Study.Among 647 African-Americans, rs11126184 in the pleckstrin (PLEK) gene was associated with OSA while rs7030789 in the lysophosphatidic acid receptor 1 (LPAR1) gene was associated with AHI using a chip-wide significance threshold of p-value<2×10⁻⁶. Among 2,904 individuals of European ancestry, rs1409986 in the prostaglandin E2 receptor (PTGER3) gene was significantly associated with OSA. Consistency of effects between rs7030789 and rs1409986 in LPAR1 and PTGER3 and apnea phenotypes were observed in independent clinic-based cohorts.Novel genetic loci for apnea phenotypes were identified through the use of customized gene chips and meta-analyses of cohort data with replication in clinic-based samples. The identified SNPs all lie in genes associated with inflammation suggesting inflammation may play a role in OSA pathogenesis.