Uric acid can enhance MAPK pathway-mediated proliferation in rat primary vascular smooth muscle cells via controlling of mitochondria and caspase-dependent cell death.

Hyperuricemia may be a risk factor for cardiovascular diseases such as hypertension and atherosclerosis, but the mechanisms underlying uric acid-induced pathological conditions remain unknown. In this study, we investigated the effect of short time and long-term administration of increasing uric acid concentrations on cell viability, proliferative and apoptotic pathways in vascular smooth muscle cells (VSMCs). Cell viability/proliferation was determined with WST-1 assay. Expression levels of mitogen-activated protein kinases (MAPKs) (phosphorylated (p)-p38 and p-p44/42 MAPK), extrinsic (caspase 3, caspase 8), and intrinsic (B-cell lymphoma-extra-large (Bcl-xL)) apoptotic pathway proteins were measured by Western blotting. In order to assess the proliferative effects of uric acid incubations on VSMCs, we monitored the proliferative/apoptosis signaling pathways for up to 24 h. Our results indicated that uric acid increases cell viability at time and dose-dependently in VSMCs. Immunoblotting results showed that uric acid treatment elevated the expression level of p-p38 MAPK but did markedly reduce the protein levels of p-p44/42, compared with all the uric acid doses-treated VSMCs, especially at 1 h. Uric acid stimulation increased caspase-3 protein levels and decreased Bcl-xL, but did not alter caspase-8 protein expression at the same dose and time. Furthermore, low uric acid incubations (0-7.5 mg/dL) did not affect any signaling pathways for long time points (6-24 h). In conclusion, our study demonstrates for the first time that VSMCs induced with uric acid can affect cell viability, proliferative, and apoptosis pathways at the widest time and dose range. These findings provide a better understanding of the uric acid effects related to vascular impairments.

Nationwide study of Escherichia coli producing extended-spectrum ß-lactamases TEM, SHV and CTX-M in Turkey.

Four hundred and forty extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli isolates were collected from 10 different hospitals in Turkey between 2011 and 2012. Clinical specimens consisted of urine (80.45%), blood (6.59%), cerebrospinal fluid (1.13%), pleural fluid (2.95%), wound (4.31%) and sputum (4.54%). ESBL-coding genes (CTX-M1, CTX-M2, TEM, SHV) were detected by PCR. According to the PCR and sequencing results, CTX-M1 was the most prevalent ß-lactamase 83.18% (366/440), followed by TEM 44.09% (194/440), CTX-M2 31.81% (140/440) and SHV 1.81% (8/440). Sequencing results showed that TEM and SHV types were TEM-1b and SHV-11, respectively. Rate of the strains harboring only CTX-M1, CTX-M2, TEM-1b and SHV-11 were 30.90%, 3.63%, 2.27% and 0.23%, respectively. Rate of the strains harboring the combinations of CTX-M1-CTX-M2, CTX-M1-CTX-M2-TEM-1b, CTX-M2-TEM-1b, CTX-M1-TEM-1b, CTX-M1-CTX-M2-TEM-1b-SHV-11, CTX-M1-TEM-1b-SHV-11, CTX-M1-SHV-11, CTX-M1-CTX-M2-SHV-11, CTX-M2-SHV-11, CTX-M2-TEM-1b-SHV-11, TEM-1b-SHV-11 were 12.95%, 11.59%, 2.95%, 26.13%, 0.45%, 0.68%, 0.22%, 0.22%, 0%, 0% and 0%, respectively. This is a nationwide study of ESBL-producing E. coli in Turkey. These results shows that CTX-M1 group is the most common type of class A ß-lactamases among ESBL-producing E. coli strains in Turkey.

[Comparison of conventional methods and real-time multiplex polymerase chain reaction for identification and typing of Brucella isolates of human origin]. / Insan Kaynakli Brucella izolatlarinin tanimlama ve tiplendirmesinde konvansiyonel yöntemler ile gerçek zamanli multipleks polimeraz zincir reaksiyonunun karsilastirilmasi.

Brucellosis which is a worldwide zoonotic disease, still constitutes a major public health problem in rural areas of Turkey. The aim of the present study was to evaluate the species and biovar distribution of 187 presumptive Brucella strains isolated from patients inhabiting at the provinces in Eastern, South Eastern and Mediterranean regions over a 7-years period (from 2001 to 2007) and to compare multiplex real-time-polymerase chain reaction (M-RT-PCR) and conventional biotyping for the differentiation of three Brucella species. The isolates were identified at genus level by conventional microbiological methods and classified using the classical Brucella species biotyping scheme based on CO2 requirement for growth, urease activity, H2S production, sensitivity to basic fuchsin and thionin (20 and 40 µg/ml), lysis by Tbilisi and Berkeley phages, and agglutination with monospecific antisera for A and M antigens. All Brucella isolates were identified as Brucella melitensis biovar 3. M-RT-PCR assay targeted bcsp31 gene and the specific integration of IS711 elements within the genome of the respective Brucella species. For the identification of Brucella spp. The primers and probes which targeted the bcsp31 gene were used. The Brucella abortus primers and probe set targeted the specific insertion of an IS711 element downstream of the alkB gene, whereas the B.melitensis primers and probe set targeted the insertion of an IS711 element downstream of BMEI1162. M-RT-PCR results were found to be 100% compatible with the reference conventional typing methods. Due to its high sensitivity, the M-RT-PCR assay could be a valuable tool for the rapid detection and differentiation of Brucella species in clinical samples which usually have very low bacterial load. These findings indicated that B.melitensis biovar 3 was by far the most frequent species for human brucellosis in these specific regions of Turkey and multiplex-RT-PCR seemed to be promising in the detection and differentiation of Brucella species.

Multiple-locus variable-number tandem-repeat analysis genotyping of human Brucella isolates from Turkey.

A multiple-locus variable-number tandem-repeat analysis (MLVA) was applied to investigate the epidemiological relationship and genetic diversity among 162 human Brucella isolates collected from all geographic regions of Turkey in an 8-year period (2001 to 2008). The isolates were genotyped by using an MLVA assay developed in Orsay, France (MLVA-16(Orsay)) including eight minisatellite (panel 1) and eight microsatellite (panel 2, subdivided into 2A and 2B) markers. Panels 1 and 2A distinguish 14 genotypes; two of these represented 85% of the strains. Panel 2B displayed a very high discriminatory power. Three loci from panel 2B had diversity index values higher than 0.74. MLVA-16(Orsay) yielded 105 genotypes; 73 were represented by a unique isolate, and 32 included two to eight isolates. The isolates from different patients within the same outbreak or from the same patient before first-line therapy and after relapse showed identical genotypes. A number of MLVA genotypes appeared to be partially restricted to some geographic areas and displayed no annual variation, possibly reflecting persistence of genotypes in certain areas for a time span of at least a decade. This study, representing the first molecular typing results of human Brucella isolates from Turkey, indicated that Turkish human Brucella melitensis isolates were most closely related to the neighboring countries' isolates included in the East Mediterranean group.