Prostaglandin E2 (PGE2) synthesis pathway is involved in coronary artery stenosis and restenosis.

The inflammatory events related to prostaglandins may play an important role in the progression of vessel stenosis. The aim of this study was to investigate the monocyte PTGES and 15-PGDH gene expression levels and the serum 13,14-dihyro-15-keto-PGF2α value involved in PGE2 metabolism in patients with coronary artery stenosis and restenosis. Moreover, the effects of miR-520, miR-1297 and miR-34 were studied on the gene expression levels. A total of sixty subjects referred for coronary angiography including healthy controls (stenosis <5%), subjects with stent no restenosis) SNR, stenosis <5%) and subjects in stent restenosis (ISR, restenosis >70%) were participated in the study. The gene expression levels and the serum 13,14-dihyro-15-keto- PGF2α value were measured by RT-qPCR and ELISA techniques, respectively. Moreover, the effects of miRNAs on the gene expression levels were investigated by the monocyte transfection of miR/PEI complexes. The PTGES and 15-PGDH gene expression levels and serum 13,14-dihyro-15-keto- PGF2α value increased significantly (P <0.05). Based on the miR-520 and miR-34 expression levels, the miR/PEI transfection studies were confirmed significantly the gene expression changes. The monocyte PGE2 synthesis pathway is actively considered in the SNR and ISR patients and might be related to miR-34 and miR-520 functions.

COX and PTGDS Gene Expression Levels in PGD2 Synthesis Pathway are Correlated with miR-520 in Patients with Vessel Restenosis.

BACKGROUND: The vessel restenosis is related to the inflammatory events in subendothelial space. It is proposed that the inflammatory agents affect the prostaglandin synthesis pathway. In this study, we investigated the COX-1, COX-2, PTGDS and miRNA-520a-5p expression levels and the serum 15-Deoxy-Δ12,14-PGJ2 metabolite values in patients with the stenosed and re-stenosed vessels. Furthermore, the associations between genes and miR-520 were evaluated in the monocyte transfection studies. METHODS: The subjects (n=60) were included three groups; healthy subjects (control (stenosis < 5%), stent no restenosis (SNR, restenosis < 5%) and in-stent restenosis (ISR, restenosis > 70%)). The miRNA and gene expression levels were measured by RT-qPCR technique. 15-Deoxy-Δ12,14-PGJ2 values were measured by the ELISA technique. The miR-520 was transfected into myocytes using PEI polymer. RESULTS: The monocyte COX-1, COX-2 and PTGDS gene expression levels and the serum 15-Deoxy- Δ12,14-PGJ2 values increased significantly in the patients. Furthermore, the miR-520 correlated conversely with the COX-1, and PTGDS gene expression levels. CONCLUSION: The results showed that the PGD2 synthesis pathway is active in the patients and, miR- 520 may be involved in the function of this pathway.

A Peptide Construct Mediates Focal Adhesion Pathway Through the Activation of Integrin Receptor.

BACKGROUND: The integrin family receptors stimulate the cellular proliferation and migration through the focal adhesion pathway by the activation of PTK2, VASP and TSP1 proteins. The purpose of this study was to investigate the integrin-ligated motifs through the activation of focal adhesion pathway. METHODS: A chimeric peptide was predicted from the integrin-mediated ligands by bioinformatics tools. The VSMCs were treated with the chimeric peptide and simvastatin. The PTK2, VASP and TSP1 protein and gene expression levels were measured by RT-qPCR and Western Blotting techniques, respectively. AutoDock Tools were used for the docking technique. RESULTS: The PTK2, VASP and TSP1 protein expression levels increased significantly in the VSMCs treated with chimeric peptide in conversely with the effects of simvastatin. The docking results suggested two motifs in the chimeric peptide. CONCLUSION: In conclusion, the chimeric peptide activated the focal adhesion pathway. The motifs 1 and 2 may be directly involved in the transduction of signal by integrin family receptors.

Strain differentiation of 13 indigenous Mycobacterium bovis isolates from infected cattle by restriction fragment length polymorphism analysis.

Mycobacterium bovis is mainly detected in cattle throughout the world. This bacterium is considered the main causative agent of tuberculosis in man and animals. M. bovis is also reported to be endemic in badgers and in farmed and feral deer. The disease caused by M. bovis is a slow progressive disease with clinical signs not apparent until late in the disease process. key factors for effective control of tuberculosis includes rapid detection, adequate therapy, and contact tracing to halt further transmission. However, in order to locate the source and route of transmission of M. bovis infection, a thorough epidemiological analysis is routinely carried out, that could lead to the control of disease in the herds and avoid economic losses. Recent developments in DNA technology and molecular biology have led to methods for the rapid detection of mycobacterium DNA. Among a number of described molecular methods, IS6110 fingerprinting is the recommended standard primary genotyping method and the most widely used worldwide. In this study, we used restriction fragment length polymorphism analysis with probes derived from the insertion element IS6110, the polymorphic GC-rich sequence (PGRS), and the direct repeat (DR) sequence which proved to be a useful method for differentiating M. bovis strains. A total of 13M. bovis samples from infected cattle in the West Azerbaijan Province of Iran were included in the study. The samples were submitted to the Tuberculosis Reference Laboratory at the Razi Vaccine and Serum Research Institute, Karaj. All isolates were cultivated on Lowenstein Jensen media with pyruvate (pyruvic acid), and then identified according to The World Organization for Animal Health (OIE) recommendations. The extracted genomic DNA samples of the isolates in the study were subjected to IS6110 primers, digested with restriction enzyme PvuII, and hybridized by oligonucleotide probes PGRS and DR. Polymorphic banding patterns obtained after hybridization discriminated the M. bovis strains and a database of strain types was established. Based on our results, the 13 isolates showed five different DNA patterns with a PGRS probe and similarly five patterns were obtained with the DR probe. PP-1 pattern was found almost among all the isolates while a distinct DNA pattern PP-3 was seen specifically from the West Azerbaijan Province.