ABSTRACT
Background: Biosynthetic teriparatide [1-34] [TPD] is a N-terminally truncated version of human parathyroid hormone [hPTH]. The recombinant form of this polypeptide has been expressed in Escherichia coli [E. coli] and approved as the first anabolic treatment of osteoporosis in the EU and the USA. Feasibility of expression and secretion of a tag- fused form of TPD into Bacillus subtilis [B. subtilis] was examined due to several advantages of B. subtilis over E. coli in production of recombinant proteins with pharmacological activities
Methods: A codon optimized gene containing TPD open reading frame carrying enterokinase site in its upstream was fully synthesized. According to our cloning scheme, this synthetic polynucleotide was used as a template for PCR amplification using engineered primers in such a way that a polyhistidin tag was added in frame to the upstream of the amplicon as well as two restriction sites at its ends. The resulted amplicon, a cassette containing His-tag, enterokinase site and TPD, from 5' to 3', was cloned into pTZ57R/T vector and subjected to sequencing.The cassette was then subcloned into pHT43 shuttle vector and transformed into B. subtilis. Expression of target protein was analyzed by SDS-PAGE and western blotting upon induction by IPTG
Results: The accuracy of construction of pHT43-TPD was confirmed by sequencing and restriction map analyses. SDS-PAGE and western blotting results showed that the recombinant fusion form of hPTH was successfully expressed and secreted into cytoplasm and extracellular medium
Conclusion: TPD may be successfully expressed and secreted in B. subtilis; however, optimization of expression conditions is required for enhancing target protein yield
ABSTRACT
Type 2 diabetes mellitus is a chronic illness causing considerable morbidity and mortality. Enormous advances have been made in medical care but more people are still having tendency to use herbal or alternative remedies. This study is a randomized, controlled trial on type 2 diabetic patients. The subject consisted of 60 patients divided randomly into three groups and supplemented daily with 100 mg sodium metavanadate and 660 mg zinc sulfate or placebo for six weeks. The following were checked at baseline of the study and after six weeks: Body Mass Index [BMI], Blood Pressure [BP], Fasting Blood Sugar [FBS], 2-h postprandial glucose [2hpp], Glycated hemoglobin [HbA1c], Triglyceride [TG], Total Cholesterol [TC], Low-Density Lipoproteins, and High-Density Lipoproteins. Also HbA1c, BMI and BP were measured after 12 weeks to evaluate the long-term effects of drugs. Statistical analysis was performed using SPSS 11.5. Data of continuous variables are expressed as means +/- standard deviation. Differences between groups were assessed by the paired T-test. Comparison between three groups was done by Post Hoc Tests. Mean age of patients was 51.39 +/- 8.60 years. The results of this study show a significant decrease in TG [P=0.01] and BMI [P=0.03]. After 12 weeks, there was a significant decrease in BMI [P=0.01] in Sodium metavanadate group. Due to zinc sulfate administration, significant decrease was seen in TG [P=0.005], TC [P=0.02], LDL [P=0.01] and systolic blood pressure [P=0.02]. After 12 weeks, there was a significant decrease in HbA1c [P=0.04] with zinc sulfate consumption. Consumption of zinc sulfate in type 2 diabetic patients could be effective in lipid profile. It is recommended to use another vanadium compound to achieve better results
Subject(s)
Humans , Male , Female , Zinc Sulfate/pharmacology , Lipids , Blood Glucose/drug effects , Diabetes Mellitus, Type 2 , Treatment OutcomeABSTRACT
Type 2 diabetes mellitus is a chronic, progressive illness that causes considerable morbidity and premature mortality. Vanadium is a trace mineral that has been claimed to be effective in controlling blood glucose levels in diabetic patients. A randomised placebo-controlled study was conducted to evaluate the effect of sodium metavanadate on selected biochemical markers in type 2 diabetic patients. Forty patients were enrolled and half of them received 100 mg sodium metavanadate daily for 6 weeks while the other half were placebo subjects. The mean age of the patients was 53.1 ± 8.5 years. Body mass index (BMI), blood pressure(BP), fasting blood sugar (FBS), 2-h postprandial glucose, glycated hemoglobin (HbA1C), triglyceride(TG), total cholesterol (TC), low-density lipoproteins (LDL), high-density lipoproteins (HDL) were determined before the start and at the end of the study. Levels of FBS, HbA1C, TC and LDL in the diabetic subjects decreased after six weeks on sodium metavanadate, but the differences were not statistically significant on comparing between pre- and posttrial levels. Based on the results, this study did not find sodium metavanadate of beneficial use as a form of vanadium supplementation among patients with type 2 diabetes.