Improving the Anti-Toxin Abilities of the CMG2-Fc Fusion Protein with the Aid of Computational Design

CMG2-Fc is a fusion protein composed of the extracellular domain of capillary morphogenesis protein 2 (CMG2) and the Fc region of human immunoglobulin G; CMG2-Fc neutralizes anthrax toxin and offers protection against Bacillus anthracis challenge. To enhance the efficacy of CMG2-Fc against anthrax toxin, we attempted to engineer a CMG2-Fc with an improved affinity for PA. Using the automatic design algorithm FoldX and visual inspection, we devised two CMG2-Fc variants that introduce mutations in the CMG2 binding interface and improve the computationally assessed binding affinity for PA. An experimental affinity assay revealed that the two variants showed increased binding affinity, and in vitro and in vivo toxin neutralization testing indicated that one of these mutants (CMG2-Fc(E117Q)) has superior activity against anthrax toxin and was suitable for further development as a therapeutic agent for anthrax infections. This study shows that the computational design of the PA binding interface of CMG2 to obtain CMG2-Fc variants with improving anti-toxin abilities is viable. Our results demonstrate that computational design can be further applied to generate other CMG2-Fc mutants with greatly improved therapeutic efficacy.     

The Association of Homocysteine with Metabolic Syndrome in a Community-Dwelling Population: Homocysteine Might Be Concomitant with Metabolic Syndrome

Objective

Elevated plasma total homocysteine (tHcy) and metabolic syndrome (MetS) are both associated with cardiovascular disease, but the association between tHcy and MetS is not well characterized. The aim of this study was to determine the relationship between tHcy and MetS.

Methods

To further estimate the time-dependent association of tHcy and MetS, we analyzed the tHcy level and MetS in 1499 subjects from a 4.8-year longitudinal study in Beijing, People’s Republic of China.

Results

In multiple linear regression analysis, baseline tHcy levels associated with age, BMI, SBP, DBP, LDL-C and Cr independently over 4.8-years follow-up; age, BMI, SBP, DBP and Cr were found to be associated with tHcy levels independently at the end of follow-up. Logistic regression analysis showed that there was no association between the baseline tHcy level and MetS over the 4.8-year follow-up (odds ratio (OR), 1.32; 95% confidence interval (CI), 0.79–2.19; P = 0.282); rather, there was an association only with hypertension as a MetS component (OR, 1.53; 95% CI, 1.06–2.21; P = 0.024). tHcy levels were associated with MetS at both cross-sectional baseline (OR, 1.38; 95% CI, 1.02–1.88; P = 0.038) and cross-sectional follow-up (OR, 1.60; 95% CI, 1.02–2.50; P = 0.041). The tHcy levels of MetS subjects were higher than those of non-MetS subjects at both cross-sectional baseline (19.35±7.92 µmol/L vs. 17.45±6.70 µmol/L, respectively; P = 0.001) and cross-sectional follow-up (18.95±7.15 µmol/L vs. 17.11±5.98 µmol/L, respectively; P = 0.02).

Conclusion

The tHcy level was not predictive of the incidence of MetS; however, it may be a risk factor for hypertension as a MetS component. Furthermore, tHcy levels were associated with MetS at cross-sectional baseline and follow-up, which suggests that a higher level of tHcy might be concomitant with MetS.     

SAPK/JNK regulates cdc2/cyclin B kinase through phosphorylation and inhibition of cdc25c

Cells undergo M phase arrest in response to stresses like UV irradiation or DNA damage. Stress-activated protein kinase (SAPK, also known as c-Jun N-terminal kinase, JNK) is activated by such stress stimuli. We addressed the potential effects of SAPK activation on cell cycle regulatory proteins. Activation of SAPK strongly correlated with inhibition of cdc2/cyclin B kinase, an important regulator of G2/M phase. SAPK directly phosphorylated the cdc2 regulator, cdc25c, in vitro on serine 168 (S168). This residue was highly phosphorylated in vivo in response to stress stimuli. cdc25c phosphorylated on S168 in cells lacks phosphatase activity, and expression of a S168A mutant of cdc25c reversed the inhibition of cdc2/cyclin B kinase activity by cell stress. Antibodies directed against phosphorylated S168 detect increased phosphorylation of S168 after cell stress. We conclude that SAPK regulates cdc2/cyclin B kinase following stress events by a novel mechanism involving inhibitory phosphorylation of the cdc2-activating phosphatase cdc25c on S168.     

Adaptation of gut microbiome and host metabolic systems to lignocellulosic degradation in bamboo rats

Bamboo rats (Rhizomys pruinosus) are among the few mammals that lives on a bamboo-based diet which is mainly composed of lignocellulose. However, the mechanisms of adaptation of their gut microbiome and metabolic systems in the degradation of lignocellulose are largely unknown. Here, we conducted a multi-omics analysis on bamboo rats to investigate the interaction between their gut microbiomes and metabolic systems in the pre- and post-weaning periods, and observed significant relationships between dietary types, gut microbiome, serum metabolome and host gene expression. For comparison, published gut microbial data from the famous bamboo-eating giant panda (Ailuropoda melanoleuca) were also used for analysis. We found that the adaptation of the gut microbiome of the bamboo rat to a lignocellulose diet is related to a member switch in the order Bacteroidales from family Bacteroidaceae to family Muribaculaceae, while for the famous bamboo-eating giant panda, several aerobes and facultative anaerobes increase after weaning. The conversion of bacteria with an increased relative abundance in bamboo rats after weaning enriched diverse carbohydrate-active enzymes (CAZymes) associated with lignocellulose degradation and functionally enhanced the biosynthesis of amino acids and B vitamins. Meanwhile, the circulating concentration of short-chain fatty acids (SCFAs) derived metabolites and the metabolic capacity of linoleic acid in the host were significantly elevated. Our findings suggest that fatty acid metabolism, including linoleic acid and SCFAs, are the main energy sources for bamboo rats in response to the low-nutrient bamboo diet.Subject terms: Metagenomics, Bacterial evolution     

Combined analysis of genome-wide-linked susceptibility loci to Kawasaki disease in Han Chinese

Kawasaki disease (KD) is a dominant cause of acquired heart disease in children due to frequent complicating coronary artery lesions (CALs). Genome-wide association study and linkage analysis have recently identified 6 susceptibility loci at genome-wide significance of P < 5.0 × 10^?8 in subjects of Japanese, Taiwanese and European. In present study, we analysed the variants of 6 single nucleotide polymorphisms (SNPs) in the genetic loci to investigate their potential effect on KD susceptibility and outcomes in Han Chinese population. As a result, the risk alleles of rs1801274 and rs2254546 were observed significant effect on KD with higher frequencies in 358 patients than those in 815 controls. The significant role of rs1801274, rs2857151 and rs2254546 in KD was found in the multi-variable logistic regression analysis of the SNPs. Two 2-locus and one 3-locus combinations of the SNPs showed significant effect on KD with stronger association with KD relative to comparable single SNP or 2-locus combinations. Significant susceptibility to CALs was found in KD patients with high-risk genotypes at both rs1801274 and rs2857151. The meta-analyses first revealed significant risk for CALs in KD patients carrying risk allele of rs11340705, and the association of rs28493229 with KD was not observed in the Han Chinese. In conclusion, the findings demonstrated that 5 of the 6 genetic loci influence the risk for KD and 3 of them may be involved in secondary CALs formation in Han Chinese. The additive effects of 3 multi-locus combinations on KD/CALs imply that some loci may participate together in certain unknown gene networks related to KD/CALs. Further function studies of the genetic loci are helpful for better understanding the pathophysiology of KD.     

利用PFU原生动物群落多样性快速监测北京通惠河水质

本文报道了北京通惠河原生动物群落多样性特征与永质的相互关系,其中ＰＦＵ原生动物１天的群集速度（功能参数）较好地反映了５个采样站（１个设在较干净水体温榆河作为对照站）水质的变化趋势,各站原生物群落的结构参数（种类组成,个体丰度,生物量和多样性指数）的差异同样与水质关系密切,通过与８０年代中期作者利用ＰＦＵ原生动物监测通惠河污染现状和自净效能的研究资料进行了比较,表明该河流水质目前得到明显的改善。     

沙障对流动沙丘区地表节肢动物分布及多样性的影响

沙障作为人工固沙植被恢复与保护绿洲和道路安全的一种手段,可以有效阻止沙丘移动,改变流动沙丘地表生态水文过程,进而影响节肢动物多样性及营养结构及功能变化。鉴于此,选取临泽县中部沙带为研究区,利用陷阱法系统调查了生物、物理沙障和流动沙丘区沙丘不同部位地表节肢动物分布及多样性。研究结果表明,沙障营建强烈影响了流动沙丘区地表节肢动物组成,生物和物理沙障区地表节肢动物的活动密度、物种丰富度及多样性均显著高于流动沙丘区,捕食性和其它食性节肢动物较植食性节肢动物对沙障营建的响应更为敏感。沙障还改变了沙丘地表节肢动物的分布规律,生物和物理沙障区地表节肢动物的活动密度从沙丘底部到顶部依次增加,而流动沙丘区地表节肢动物主要分布在沙丘中下部。谢氏宽漠王和尖尾东鳖甲在流动沙丘区主要分布沙丘的中下部,沙障营建导致沙丘顶部谢氏宽漠王和尖尾东鳖甲的活动密度大幅提高。此外,研究还发现生物和物理沙障对地表节肢动物的影响不同,生物沙障区地表节肢动物活动密度显著高于物理沙障区,捕食性和东鳖甲属等其它食性节肢动物的活动密度在生物沙障区均高于物理沙障区。总之,沙障营建及类型差异强烈影响地表节肢动物的分布及多样性,它通过资源上行效应和土壤物理环境的改变提高了地表节肢动物的多样性及营养结构,从而影响了土壤生态系统结构及功能变化过程。     

木麻黄根乙酸乙酯部位化学成分研究

为阐明木麻黄（Casuarina equisetifolia）的化学成分,采用有机溶剂提取、萃取及多种分离技术,从其根中分离得到14个化合物。经光波谱分析,其结构分别鉴定为5-hydroxy-3-methoxyphenyl-6-O-syringoyl-β-d-glucopyranoside （1）,高良姜苷A （2）,heterophylloside C （3）、6''-O-vanilloylisotachioside （4）、3,4,5-trimethoxyphenyl-6-O-syringoyl-β-d-glucopyranoside （5）、香草醛（6）、丁香醛（7）、3,4-二羟基苯甲醛（8）、齐墩果酸（9）、桦木酸（10）、胡萝卜苷（11）、胡萝卜苷亚油酸酯（12）、（±）-lyoniresinol 2-O-a-rhamnoside （13）和（-）-9-acetyl-isolariciresinol 9''-O-a-l-rhamnopyranoside （14）,其中化合物14为新的木脂素。     

Design, synthesis, and biological activity of potent and selective inhibitors of mast cell tryptase

A new series of novel mast cell tryptase inhibitors is reported, which features the use of an indole structure as the hydrophobic substituent on a m-benzylaminepiperidine template. The best members of this series display good in vitro activity and excellent selectivity against other serine proteases.