Stratified Communities of Active Archaea in Shallow Sediments of the Pearl River Estuary, Southern China

Marine subsurface sediments represent a novel archaeal biosphere with unknown physiology. To get to know the composition and ecological roles of the archaeal communities within the sediments of the Pearl River Estuary, Southern China, the diversity and vertical distribution of active archaea in a sediment core were characterized by 16S rRNA phylogenetic analysis of clone libraries derived from RNA. In this study, the archaeal diversity above, within, and beneath the sulfate-methane transition zone (SMTZ) in the Pearl River Estuary sediment core was described. The majority of the clones obtained from the metabolically active fraction of the archaeal community were most closely related to miscellaneous crenarchaeotal group and terrestrial miscellaneous euryarchaeotal group. Notably, although the Pearl River Estuary sediment belong to high methane and high organic carbon environment, sequences affiliated with methanotrophic and methanogenic archaea were detected as minor group in 16S rRNA clone libraries. No obvious evidence suggested that these unknown archaeal phylotypes related directly to anaerobic oxidation of methane in SMTZ. This is the first phylogenetic analysis of the metabolically active fraction of the archaeal community in the coastal sediment environments.     

Separation of supercoiled from open circular forms of plasmid DNA,and biological activity detection

To establish a cost-effective purification process for the large-scale production of plasmid DNA for gene therapy and DNA vaccination, a single anion-exchange chromatography (AEC) step was employed to purify supercoiled plasmid DNA (sc pDNA) from other isoforms and Escherichia coli impurities present in a clarified lysate. Two different size and conformation plasmids were used as model targets, and showed similar elution behavior in this chromatographic operation, in which sc pDNA was effectively separated from open circle plasmid DNA (oc pDNA) in a salt gradient. The process delivered high-purity pDNA of homogeneity of 95 ± 1.1% and almost undetectable levels of endotoxins, genomic DNA, RNA and protein, at a yield of 65 ± 8%. Furthermore, the transfection efficiency (29 ± 0.4%) was significantly higher than that (20 ± 0.1%) of a pDNA control. The present study confirms the possibility of using a single AEC step to purify sc pDNA from other isoforms and host contaminants present in a clarified E. coli lysate.     

Synthesis and characterization of selective dopamine D₂ receptor ligands using aripiprazole as the lead compound

A series of compounds structurally related to aripiprazole (1), an atypical antipsychotic and antidepressant used clinically for the treatment of schizophrenia, bipolar disorder, and depression, have been prepared and evaluated for affinity at D(?-like) dopamine receptors. These compounds also share structural elements with the classical D(?-like) dopamine receptor antagonists, haloperidol, N-methylspiperone, domperidone and benperidol. Two new compounds, 7-(4-(4-(2-methoxyphenyl)piperazin-1-yl)butoxy)-3,4-dihydroquinolin-2(1H)-one oxalate (6) and 7-(4-(4-(2-(2-fluoroethoxy)phenyl)piperazin-1-yl)butoxy)-3,4-dihydroquinolin-2(1H)-one oxalate (7) were found to (a) bind to the D? receptor subtype with high affinity (K(i) values < 0.3 nM), (b) exhibit >50-fold D? versus D? receptor binding selectivity and (c) be partial agonists at both the D? and D? receptor subtype.     

miR‐129‐2‐3p directly targets SYK gene and associates with the risk of ischaemic stroke in a Chinese population

Spleen tyrosine kinase (SYK) gene has been identified as novel susceptibility locus for ischaemic stroke (IS) previously. However, regulation of SYK gene remains unknown in IS. In this study, we aimed to identify miRNAs that might be involved in the development of IS by targeting SYK gene. miRNAs were firstly screened by bioinformatics predicting tool. The expression levels of SYK gene were detected by qRT‐PCR and western blotting, respectively, after miRNA transfection. Luciferase reporter assay was applied to investigate the direct binding between miRNAs and target gene. miRNA levels were detected by miRNA TaqMan assays in the blood cells of 270 IS patients and 270 control volunteers. Results suggest that SYK gene might be a direct target of miR‐129‐2‐3p. The blood level of miR‐129‐2‐3p was significantly lower in IS patients (P < 0.05), and negatively associated with the risk of IS (adjusted OR: 0.88; 95% CI: 0.80‐0.98; P = 0.021) by multivariable logistic regression analysis. The blood levels of SYK gene were significantly higher in IS patients, and miR‐129‐2‐3p expression was negatively correlated with mean platelet volume. In summary, our study suggests that miR‐129‐2‐3p might be involved in the pathogenesis of IS through interrupting SYK expression and the platelet function, and further investigation is needed to explore the underlying mechanism.     

Two Spx Regulators Modulate Stress Tolerance and Virulence in Streptococcus suis Serotype 2

Streptococcus suis serotype 2 is an important zoonotic pathogen causing severe infections in pigs and humans. The pathogenesis of S. suis 2 infections, however, is still poorly understood. Spx proteins are a group of global regulators involved in stress tolerance and virulence. In this study, we characterized two orthologs of the Spx regulator, SpxA1 and SpxA2 in S. suis 2. Two mutant strains (ΔspxA1 and ΔspxA2) lacking the spx genes were constructed. The ΔspxA1 and ΔspxA2 mutants displayed different phenotypes. ΔspxA1 exhibited impaired growth in the presence of hydrogen peroxide, while ΔspxA2 exhibited impaired growth in the presence of SDS and NaCl. Both mutants were defective in medium lacking newborn bovine serum. Using a murine infection model, we demonstrated that the abilities of the mutant strains to colonize the tissues were significantly reduced compared to that of the wild-type strain. The mutant strains also showed a decreased level of survival in pig blood. Microarray analysis revealed a global regulatory role for SpxA1 and SpxA2. Furthermore, we demonstrated for the first time that Spx is involved in triggering the host inflammatory response. Collectively, our data suggest that SpxA1 and SpxA2 are global regulators that are implicated in stress tolerance and virulence in S. suis 2.     

橡胶树橡胶生物合成调控相关基因表达丰度比较

蛋白质是构成生命系统的基本元件之一,是大部分生物学功能的执行者。蛋白质丰度与其生物学功能息息相关,其丰度受基因表达过程中各环节严格精密的调控。其中,蛋白质丰度与其相应mRNA丰度存在较强的相关性,蛋白质丰度差异的40%可由mRNA丰度来解释。茉莉酸信号途径调节巴西橡胶树中的天然橡胶生物合成,但相关基因彼此间的表达丰度差异尚待阐明。该文比较了S/2D d3割胶制度下,15个橡胶生物合成调控相关基因COI1、JAZ1、JAZ2、JAZ3、MYC1、MYC2、MYC3、MYC4、MYC5、GAPDH、HMGR1、SRPP、REF、HRT1、HRT2以及2个常用内参基因18S、ACTIN1在10个橡胶树种质胶乳中的表达丰度差异;将ACTIN1的表达丰度设定为1,以此为标准计算出样品中其他基因的表达丰度。结果表明:相同个体中不同基因的转录丰度差异明显,不同个体中相同基因集的丰度大小排序存在一定差异;同一基因在不同个体中的转录丰度差异明显,这16个基因的最大丰度分别是最低丰度的9.43、6.04、10.02、12.29、18.82、9.22、38.46、112.83、121.36、15.34、19.09、13.54、10.05、19.80、24.83、11.82倍,他们的变异系数分别为73.05%、55.19%、69.09%、67.37%、66.59%、53.87%、83.25%、122.02%、166.34%、59.89%、70.59%、75.67%、74.20%、68.34%、84.23%、78.59%;总的来说,在群体水平上,16个基因的转录丰度从高到低依次为18S SRPPHMGR1REFMYC2/HRT1COI1MYC1/MYC4GAPDH/JAZ1/MYC5JAZ2HRT2/MYC3/JAZ3,他们的群体平均丰度依次为ACTIN1的28 382.26、43.64、11.39、7.16、5.47、5.10、1.07、0.75、0.74、0.45、0.42、0.33、0.12、0.06、0.06、0.04倍。值得注意的是,无论在个体水平还是群体水平上,18S的丰度毫无疑问是最大的,在mRNA中,SRPP的丰度最大,JAZ1大于JAZ2和JAZ3,MYC2大于MYC1、MYC3、MYC4、MYC5,HRT1大于HRT2。综上结果表明,结构基因和功能基因的丰度高于调控基因。在基因相对表达分析中,常对目的基因和内参基因作均一化处理,从而掩盖了不同基因间的真实丰度差异,因此,在基因表达分析中,既要关注基因的相对表达量,也要关注基因间的丰度差异,这有助于更全面地理解基因的功能。     

Group A Streptococcus Secreted Esterase Hydrolyzes Platelet-Activating Factor to Impede Neutrophil Recruitment and Facilitate Innate Immune Evasion

The innate immune system is the first line of host defense against invading organisms. Thus, pathogens have developed virulence mechanisms to evade the innate immune system. Here, we report a novel means for inhibition of neutrophil recruitment by Group A Streptococcus (GAS). Deletion of the secreted esterase gene (designated sse) in M1T1 GAS strains with (MGAS5005) and without (MGAS2221) a null covS mutation enhances neutrophil ingress to infection sites in the skin of mice. In trans expression of SsE in MGAS2221 reduces neutrophil recruitment and enhances skin invasion. The sse deletion mutant of MGAS5005 (Δsse ^MGAS5005) is more efficiently cleared from skin than the parent strain. SsE hydrolyzes the sn-2 ester bond of platelet-activating factor (PAF), converting biologically active PAF into inactive lyso-PAF. K_M and k _cat of SsE for hydrolysis of 2-thio-PAF were similar to those of the human plasma PAF acetylhydrolase. Treatment of PAF with SsE abolishes the capacity of PAF to induce activation and chemotaxis of human neutrophils. More importantly, PAF receptor-deficient mice significantly reduce neutrophil infiltration to the site of Δsse ^MGAS5005 infection. These findings identify the first secreted PAF acetylhydrolase of bacterial pathogens and support a novel GAS evasion mechanism that reduces phagocyte recruitment to sites of infection by inactivating PAF, providing a new paradigm for bacterial evasion of neutrophil responses.     

Effect of beating on recycled properties of unbleached eucalyptus cellulose fiber

The effects of beating on recycled properties of eucalyptus cellulose fiber were studied by analyzing the changes of morphological parameters (fiber length and the fines content), physical properties (tensile strength, breaking length and the stretch), WRV, crystal structure of cellulose and pore structure of cellulose fiber. The results showed that beating caused the fine content increase. Tensile strength, breaking length and the stretch increased with the increasing beating time. WRV of the first cycle beaten eucalyptus pulp was increased by 32.1%, compared to the first cycle unbeaten pulp. WRV increased with the increase in beating degree. However, crystallinity of cellulose increased, and then decreased with an increase in beating degree. FTIR spectra showed that there were no drastic changes in the functional groups of the eucalyptus pulp cellulose during beating. Fiber pore size was gradually diverted into macropore with the increase in beating degree, resulting in the mean pore volume increased.