Genetic diversity of Escherichia coli isolated from commercial swine farms revealed by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and repetitive extragenic palindrome PCR (REP-PCR)

The objective of this study was to use enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and repetitive extragenic palindrome PCR (REP-PCR) for the analysis of genetic diversity among Escherichia coli strains isolated from commercial swine farms in Sichuan province of China. Thirty four strains of E. coli were selected by selective medium and conventional biochemical test from fresh stool samples of swines in five farms in Sichuan province. The isolates were identified by 160 kinds of E. coli O serums. The results show that 30 strains were determined among 34 E. coli isolates, 12 kinds of O serogroups were obtained on the basis of the agglutination test. The predominant types are O23, O113 and O120, representing 35.4%. Furthermore, the genotypes and phylogenetic relationship of all isolates were analysed by Enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and repetitive extragenic palindrome PCR (REP-PCR), 34 E. coli isolates were clustered to 19 ERIC-PCR genotypes and 13 REP-PCR genotypes. The isolates from the same farm or sharing the same serotyping showed different genotype. And the isolates which could not be serotyped were genotyped by ERIC-PCR and REP-PCR. The analysis of genetic type and original source revealed that isolates from different farms had different genetic types. The subtypes of E. coli are also different within a single farm. Genetic variability with E. coli strains isolated from swine farms in China has been demonstrated. The presence of ERIC-PCR and REP-PCR sequences in the genome of E. coli was confirmed. ERIC-PCR and REP-PCR techniques are more rapid methods for molecular typing of E. coli strain. They are also useful methods for diversity survey of E. coli and the two methods analyzes genetic diversity of E. coli isolated in Sichuan of China.Key words: Escherichia coli, serotype, enterobacterial repetitive intergenic consensus PCR (ERIC-PCR), repetitive extragenic palindrome PCR (REP-PCR)

The N-Terminal DH-PH Domain of Trio Induces Cell Spreading and Migration by Regulating Lamellipodia Dynamics in a Rac1-Dependent Fashion

The guanine-nucleotide exchange factor Trio encodes two DH-PH domains that catalyze nucleotide exchange on Rac1, RhoG and RhoA. The N-terminal DH-PH domain is known to activate Rac1 and RhoG, whereas the C-terminal DH-PH domain can activate RhoA. The current study shows that the N-terminal DH-PH domain, upon expression in HeLa cells, activates Rac1 and RhoG independently from each other. In addition, we show that the flanking SH3 domain binds to the proline-rich region of the C-terminus of Rac1, but not of RhoG. However, this SH3 domain is not required for Rac1 or RhoG GDP-GTP exchange. Rescue experiments in Trio-shRNA-expressing cells showed that the N-terminal DH-PH domain of Trio, but not the C-terminal DH-PH domain, restored fibronectin-mediated cell spreading and migration defects that are observed in Trio-silenced cells. Kymograph analysis revealed that the N-terminal DH-PH domain, independent of its SH3 domain, controls the dynamics of lamellipodia. Using siRNA against Rac1 or RhoG, we found that Trio-D1-induced lamellipodia formation required Rac1 but not RhoG expression. Together, we conclude that the GEF Trio is responsible for lamellipodia formation through its N-terminal DH-PH domain in a Rac1-dependent manner during fibronectin-mediated spreading and migration

Targeting RNS/caveolin-1/MMP signaling cascades to protect against cerebral ischemia-reperfusion injuries: potential application for drug discovery

Reactive nitrogen species (RNS) play important roles in mediating cerebral ischemia-reperfusion injury. RNS activate multiple signaling pathways and participate in different cellular events in cerebral ischemia-reperfusion injury. Recent studies have indicated that caveolin-1 and matrix metalloproteinase (MMP) are important signaling molecules in the pathological process of ischemic brain injury. During cerebral ischemia-reperfusion, the production of nitric oxide (NO) and peroxynitrite (ONOO-), two representative RNS, down-regulates the expression of caveolin-1 (Cav-1) and, in turn, further activates nitric oxide synthase (NOS) to promote RNS generation. The increased RNS further induce MMP activation and mediate disruption of the blood-brain barrier (BBB), aggravating the brain damage in cerebral ischemia-reperfusion injury. Therefore, the feedback interaction among RNS/Cav-1/MMPs provides an amplified mechanism for aggravating ischemic brain damage during cerebral ischemia-reperfusion injury. Targeting the RNS/Cav-1/MMP pathway could be a promising therapeutic strategy for protecting against cerebral ischemia-reperfusion injury. In this mini-review article, we highlight the important role of the RNS/Cav-1/MMP signaling cascades in ischemic stroke injury and review the current progress of studies seeking therapeutic compounds targeting the RNS/Cav-1/MMP signaling cascades to attenuate cerebral ischemia-reperfusion injury. Several representative natural compounds, including calycosin-7-O-β-D-glucoside, baicalin, Momordica charantia polysaccharide (MCP), chlorogenic acid, lutein and lycopene, have shown potential for targeting the RNS/Cav-1/MMP signaling pathway to protect the brain in ischemic stroke. Therefore, the RNS/Cav-1/MMP pathway is an important therapeutic target in ischemic stroke treatment.published_or_final_versio