Vascular miR‐181b controls tissue factor‐dependent thrombogenicity and inflammation in type 2 diabetes

BACKGROUND: Diabetes mellitus is characterized by chronic vascular inflammation leading to pathological expression of the thrombogenic full length (fl) tissue factor (TF) and its isoform alternatively-spliced (as) TF. Blood-borne TF promotes factor (F) Xa generation resulting in a pro-thrombotic state and cardiovascular complications. MicroRNA (miR)s impact gene expression on the post-transcriptional level and contribute to vascular homeostasis. Their distinct role in the control of the diabetes-related procoagulant state remains poorly understood. METHODS: In a cohort of patients with poorly controlled type 2 diabetes (n = 46) plasma levels of miR-181b were correlated with TF pathway activity and markers for vascular inflammation. In vitro, human microvascular endothelial cells (HMEC)-1 and human monocytes (THP-1) were transfected with miR-181b or anti-miR-181b and exposed to tumor necrosis factor (TNF) α or lipopolysaccharides (LPS). Expression of TF isoforms, vascular adhesion molecule (VCAM) 1 and nuclear factor (NF) κB nuclear translocation was assessed. Moreover, aortas, spleen, plasma, and bone marrow-derived macrophage (BMDM)s of mice carrying a deletion of the first miR-181b locus were analyzed with respect to TF expression and activity. RESULTS: In patients with type 2 diabetes, plasma miR-181b negatively correlated with the procoagulant state as evidenced by TF protein, TF activity, D-dimer levels as well as markers for vascular inflammation. In HMEC-1, miR-181b abrogated TNFα-induced expression of flTF, asTF, and VCAM1. These results were validated using the anti-miR-181b. Mechanistically, we confirmed a miR-181b-mediated inhibition of importin-α3 (KPNA4) leading to reduced nuclear translocation of the TF transcription factor NFκB. In THP-1, miR-181b reduced both TF isoforms and FXa generation in response to LPS due to targeting phosphatase and tensin homolog (PTEN), a principal inducer for TF in monocytes. Moreover, in miR-181-/- animals, we found that reduced levels of miR-181b were accompanied by increased TF, VCAM1, and KPNA4 expression in aortic tissue as well as increased TF and PTEN expression in spleen. Finally, BMDMs of miR-181-/- mice showed increased TF expression and FXa generation upon stimulation with LPS. CONCLUSIONS: miR-181b epigenetically controls the procoagulant state in diabetes. Reduced miR-181b levels contribute to increased thrombogenicity and may help to identify individuals at particular risk for thrombosis

MicroRNA-19a contributes to the epigenetic regulation of tissue factor in diabetes

Background: Diabetes mellitus is characterized by chronic vascular disorder and presents a main risk factor for cardiovascular mortality. In particular, hyperglycaemia and inflammatory cytokines induce vascular circulating tissue factor (TF) that promotes pro-thrombotic conditions in diabetes. It has recently become evident that alterations of the post-transcriptional regulation of TF via specific microRNA(miR)s, such as miR-126, contribute to the pathogenesis of diabetes and its complications. The endothelial miR-19a is involved in vascular homeostasis and atheroprotection. However, its role in diabetes-related thrombogenicity is unknown. Understanding miR-networks regulating procoagulability in diabetes may help to develop new treatment options preventing vascular complications. Methods and results: Plasma of 44 patients with known diabetes was assessed for the expression of miR-19a, TF protein, TF activity, and markers for vascular inflammation. High miR-19a expression was associated with reduced TF protein, TF-mediated procoagulability, and vascular inflammation based on expression of vascular adhesion molecule-1 and leukocyte count. We found plasma expression of miR-19a to strongly correlate with miR-126. miR-19a reduced the TF expression on mRNA and protein level in human microvascular endothelial cells (HMEC) as well as TF activity in human monocytes (THP-1), while anti-miR-19a increased the TF expression. Interestingly, miR-19a induced VCAM expression in HMEC. However, miR-19a and miR-126 co-transfection reduced total endothelial VCAM expression and exhibited additive inhibition of a luciferase reporter construct containing the F3 3′UTR. Conclusions: While both miRs have differential functions on endothelial VCAM expression, miR-19a and miR-126 cooperate to exhibit anti-thrombotic properties via regulating vascular TF expression. Modulating the post-transcriptional control of TF in diabetes may provide a future anti-thrombotic and anti-inflammatory therapy

Study of the Effective Parameters on the Making Use of Protective Devices

<p><strong>Background and objectives</strong></p><p>Noise threats health of many groups of industrial workers and causes hearing loss. Use of personal protective device is the best control method to protect against hazardous conditions. Hence, this investigation was carried out to determine situation of using of protective devices and effective parameters on it, in Qom province workers community in 2006.</p><p> </p><p><strong>Methods</strong></p><p>This research is descriptive-sectional study. Sample volume was designed 378 persons working in factories in Qom. First of all, list of Qom factories with noise pollution problems, were collected and 30 important factories among them were selected randomly. In the second stage, 378 persons were selected randomly from workers. The interest information was obtained by questionnaire and collected data were analyzed by SPSS software.</p><p> </p><p><strong>Results</strong></p><p>The obtained results showed that, 83.6% of workers have been using ear protective devices. 296 of them, which were using ear protective devices, had an occupational hygienist in their workplaces. This research also showed that, 109 workers that used ear protective devices, had moderate knowledge level. Moreover, 82.5% of trained workers have used ear protective devices. The statistical analysis of the results showed that there were no significant relationship between use of ear protective devices and existence of occupational hygienist in workplace, knowledge and age of workers, worker’s antecedent, physical health of workers and kind of ear protective devices (p> 0.05). </p><p> </p><p><strong>Conclusion </strong></p><p>These results showed that among all considered parameters; only four parameters were effective in using ear protective devices; education of workers before employment, head workman and employer’s knowledge level, factories facilitation and kind of ear protective devices.</p><p> </p&gt

The effect of Progressive muscle relaxation on the Depression in patients with type 2 diabetes

Introduction:Depression is one of the most common psychiatric disorders in patients with diabetes that its incidence in addition to the impact on the course of treatment, can be changed the prognosis. This study aimed to determine the effects of progressive muscle relaxation on depression in patients with type 2 diabetes. Methods &amp; Materials: This study was a clinical trial that conducted on 77 patients who referred to Diabetes center of Sabzevar city. The participants completed the Beck Depression Inventory and if they earned 14-28 (mild to moderate depression), included to study. Then, participants after consent, were divided Randomly into two groups of progressive muscle relaxation (n= 37) and control (n= 40).relaxationperformed 2 times a week for 20-30 minuteswithin 6 weeks, under the supervision of a researcher. Beck Depression Inventory was completed at the end of the sixth week again.In analytic measurements of this study used of statistical analysis in SPSS software (Version 16). A P value of < 0.05 was considered to be statistically significant. Results: The mean depression score after intervention in relaxation group compared to control group was ranged from 20.91±4.20 to 11.29±2.85 and from 20.60±4.21 to 19.99±3.61, respectively.According to independent t-test, there was significant differencebetween case and control groups (P<0/001). Paired t-test showed statistically significant difference in terms of depression scores before and after intervention(P<0/001); while in control group had not any statistical significancy (p=0.204) Conclusion:Progressive Muscle Relaxation is effective way to reduce depression in patients with diabetes type 2

Cloning and expression of Brucella outer membrane protein 36kDa (OMP2b) in E. coli

Background & Objective: Brucellosis is an important zoonotic disease of economic significance. Brucella species are gram-negative, facultative intracellular bacteria, and are capable of replicating in the phagosomes of macrophages. They cause infection in several animal species and humans. Prevention of new diseases and diagnosis of cases infected with the organism are both essential for eradication of the disease. Characterization and evaluation of different antigens of Brucella cells has a key role in progression of prevention and diagnosis programs. Here, we report the production and purification of recombinant 31kDa outer membrane protein Brucella abortus (Omp2b). Materials & Methods: Brucella abortus 36kDa outer membrane protein gene was amplified with PrimeSTAR® HS DNA polymerase, cloned in pJET1.2. The target gene was subcloned in pET28a (+). Recombinant pET28a vectors were transformed into E coli BL21 (DE3). Expression of recombinant protein was induced with 1mM IPTG. Proteins were absorbed to Ni-NTA agarose resins and Recombinant proteins were eluted with 250mM imidazol. Imidazol removed by dialysis. Proteins were assayed by Western-blotting and rOmp2b was probed by Brucella rabbit anti serum. Result: Appearance of a golden brown band at the site of reaction, in Western blotting confirmed successfully clone and expression. We purified Omp2b by affinity chromatography and this method prepared refolds proteins on the column. Conclusion: Omp2b were successfully cloned, expressed and purified. The recombinant proteins were recognized by polyclonal antiserum which suggests the accuracy of procedure

Study of the Antifungal Activity of Shikonin and Alcoholic – Oily Extracts of Iranian Arnebia euchroma L

Introduction: Today regarding drug resistance of fungi and bacteria, many researches are focused on herbal-based medication. As these herbal-based medications can show better adaptivity, the minimum advantage of them compared to synthetic drugs is that they are harmless. This article aimed to study the antifungal effect of alcoholic extract and essence of Arnebia euchroma L (Abukhalsa) roots on saprophytic and dermatophytic fungi. Methods: In this research, the roots were collected from Zagros heights in spring. Then they were dried and 300 mL ethanol was added to each 100 g dried powder. The alcoholic extraction was performed by maceration and the extract was concentrated by distillation in vacuum. The clevenger apparatus was used to extract the essence; then it was extracted by boiling water at vacuum for 4–6 hours. Shikonin was provided in commercial form. The antifungal activities of alcoholic extract, essence and Shikonin were studied and recorded using cylinder test based on the diameter of inhibition zone in Sabouraud-Dextrose agar. Minimum inhibitory concentration (MIC) and Minimum fungicidal concentration (MFC) were measured by broths macrodilution tests. Results: The results from cylinder, MIC and MFC tests showed that 30% of shikonin was more effective than root on fungi. Our data demonstrated that alcoholic extract was better than oily extract. Conclusion: The alcoholic extract had better characteristics than the essence. To confirm the final findings, further researches are required

Relationship between Calcium, Phosphorous and Parathoromone with Calcification of Heart Valves In Dialyzed Patients In Qom, 1385-1386

Background and ObjectivesMetastatic calcification is usually found in patients with chronic renal failure and hemodialyzed patients. Heart calcification becomes worse in hemodialyzed patients due to several factors. The aim of this study was to investigate relationship between Calcium, Phosphorous and Parathoromone(PTH) with calcification of heart valves in dialyzed patients. MethodsThis study is a Case- series study. 71 patients with calcification of heart valves who have referred for dialysis to Kamkar and Valiasr hospitals in Qom were selected as study population through echocardiography. Levels of Calcium, Phosphorous and PTH in the patients were assayed by suitable methods.ResultsOur results showed that in %17/5 of study population the level of calcium was higher than 10.4mg/dl, in %75.1 the level of PTH was higher than 70um/lit and finally the level of Phosphorous in %54.2 was higher than 4.5mg/dl.We concluded that there are a meaningful relationship between calcification of heart valves and levels of calcium, Phosphorous and PTH.ConclusionMeasurement of serum calcium, Phosphorous and PTH and performing echocardiography for prevention of calcification of heart valves are necessary in individuals who are dialyzed.Keywords: Heart Valves Calcification, Calcium, PTH, Phosphorou

The combined use of recombinant and pVAX-omp31 DNA Vaccine for immunological protection against pathogenic Brucellas melitesnsis in an experimental model of BALB/c Mice

Background and Aim: Brucellosis is still an important zoonotic infection and evaluation of immunologic properties of various bacterial antigens along with different vaccination strategies helps in designing efficient vaccines against the disease. The aim of this study is to immunological evaluate the eukaryotic vector pVAX1, carrying the outer membrane protein gene of 31 kDa (Omp31) B.melitensis. Materials and Methods: In this study which was carried out in 2014, whole sequence of omp31 of B. melitensis was inserted between BamHI and XhoI of pVAX1 plasmid vector. Female BALB/c mice aged 6-8 weeks (purchased from Pasteur Institute of Iran) were immunized intra-muscularly with 100μg of the construct, followed by either protein or plasmid boosters separately. The level of IL-4, IL-12, IFN-γ, total serum IgG, and specific IgG1 and IgG2a against recombinant Omp31 were evaluated. Finally the protective immune response following exposure to B. melitensis 16M was evaluated. Results: DNA-vaccine omp31 career with protein reminders Omp31, stimulate higher levels of IFN-γ, IL-12 and IgG2a compared to groups of DNA-vaccine or recombinant protein. Protective immunity was also significantly higher in mice which immunized with DNA vaccine– protein regimen. Conclusions: Mice which immunized with DNA vaccine–protein regimen showed a significantly higher levels of IL-12 and IFN-γ along with serum IgG2a which together imply augmentation of T cell-mediated immune responses against Omp31. The latter was confirmed by significant protective response to B. melitensis 16M challenge