real time PCR

Introduction: Bone formation is disturbed in type 1 diabetes followed by changes in the bone microstructure. The most important metabolic disorder in diabetes is osteoporosis, which is characterized by bone loss and bone structure degradation. This study aimed to determine the effect of low-power laser on bone defect repair in the experimental model of diabetes and osteoporosis.

Materials & Methods: A total of 30 four-month-old female Wistar rats weighing 190-220 g were selected and randomly divided into six groups, including 1: non-diabetic control (Co.), 2: non-diabetic laser (L.), 3: diabetic control (Co.D.), 4: diabetic laser (L.D.), 5: diabetic alendronate (A.D.), and 6: diabetic laser + alendronate (L.A.D.). Diabetes was induced in groups 3, 4, 5, and 6. All groups underwent ovariectomy and partial bone defect. In the laser group, a low-level laser (890nm, 80 Hz, 1/5J / cm 2) was radiated to 3 points at the defect location. Tibia bones were collected, and Real-time PCR was performed after a month. The data were analyzed using ANOVA. A p-value less than P<0.05 was considered statistically significant. Ethics code: 13237-91-1-1393-10397

Findings: The t-test showed a significant decrease in tibia bone density in diabetic and osteoporotic rats, compared to the non-diabetic control group. Moreover, analysis of gene expression data (ANOVA, P<0.05) revealed a significant difference between the group of diabetic laser + alendronate and other groups in terms of Runx2 gene expression and Osteocalcin.

Discussions & Conclusions: According to the findings, laser therapy combined with alendronate can accelerate the repair of partial bone defect in the experimental model of diabetes and osteoporosis.

Background and Objective: Streptomyces are gram-positive and aerobic bacterial strains that are isolated from different sources. Streptomyces have the ability to produce secondary metabolites and biologically active substances and are therefore very important in the field of biocontrol. Urate oxidase is a microbial enzyme product that can be extracted from a variety of sources, including streptomycin. In the present study, cloning of the urate oxidase gene isolated from seawater streptomycosis was performed in Escherichia coli Origami bacteria.
Methods: In this descriptive study, a total of 60 water and sediment samples were collected from different depths of the Caspian Sea coast in Mazandaran province, Iran. The Geram, staining methyl red, VP, citrate, starch hydrolysis, casein hydrolysis, nitrate reduction, oxidase and catalase tests were performed to identify and isolate Streptomyces. The urate oxidase gene was cloned using the T-A cloning method using the PTG-19 vector inside the host of Escherichia coli Origami. The expression of cloned genes in recombinant colonies was investigated by Real-Time PCR. The phylogenetic tree was drawn using clustalX and Mega5 software.
Results: Screening of marine water samples identified 12 isolated streptomyces, all of which had the urate oxidase gene. The expression of urate oxidase gene in Escherichia coli Origami was confirmed by Real-Time PCR. The results of phylogenetic studies identified some close relatives of Streptomyces as candidates for subsequent studies.
Conclusion: Streptococcus bacteria can be considered as a rich source of secondary metabolites with many applications and can be used as a native to produce the enzyme urate oxidase. By using different cloning hosts and examining optimal production conditions, this strain can be a candidate for future studies to develop antimicrobial drugs and compounds.
Keywords: Cloning [MeSH], Urate Oxidase [MeSH], Real Time PCR [MeSH], Caspian Sea [MeSH],