ABSTRACT
Abstract Flatulence and fullness of stomach is one of the most common problem associated with chickpea primary due to presence of some oligosaccharides and phenols. In this investigation Desi and Kabuli varieties were compared for these oligosaccharides and phenolic compounds. Furthermore, the effect of different processing and cooking methods such as soaking, cooking and germination in the reduction of these antiphysiological factors were are also studies. Maximum tannic acid (0.90 ± 0.20%) was observed in Parbat and C-44 while minimum (0.60 ± 0.04%) in Karak-2. Stachyose contents ranged between 1.10 ± 0.05 (Karak-3) to 1.42 ± 0.02% (Parbat) while raffinose was 0.63 ± 0.05(Karak-3) to 0.81 ± 0.02% (Dasht). The highest tannic acid content was reduced up to 50% in C-44 by cooking of 72 hours germinated seeds. Stachyose and raffinose contents were completely removed after 72 hours germination. Present studies revealed that cooking after germination is the most effective method to reduce the anti-nutritional factors of chickpea. Individually, soaking and cooking also contributed to the loss of the same factors but to a lesser extent.
ABSTRACT
Background: ACE a renin-angiotensin system that regulates blood pressure, balance of fluids and salts in body and PAI-1 is a serine protease inhibitor, which inhibits tissue plasminogen activator andurokinase.They are thought to play an important role in pathophysiology of kidney disease in diabetes. Aim: In our present study, we studied the association of altered ACE-gene and PAI-1 gene with diabetic retinopathy (DR) and NDR in 592 samples consisted of (cohort I; 196 DR patients, cohort II; 200 diabetic nonretinopathy (DNR) and cohort III, 196 respective controls. Methods: For genotyping of ACE-gene and PAI-1 gene, genomic DNA was isolated and purified which was then amplified by PCR, and thePCR products analyzedwere by Agarose gel electrophoresis. Results: In first part, the ACE genotype and allele frequency distribution was studied. For ACE gene polymorphism, the genotype and allele frequency distribution were analyzed in DR subjects and respective controls. The results indicated that there is no statistically significant difference between DR males and females compared to respective controls. The results were significantly high between genotype frequencies of DR and DNR in males. The recessive model was found to be significantly associated with the DR male subjects (OR=0.45 [95% CI=0.20-0.99], p<0.05), whereas in females these are non-significant as compared to respective controls individuals. In second part of study, the disease status analysis of ACE gene on basis of DR stages (NPDR and PDR) was observed. The χ2 analysis indicated that results are significantly different between NPDR and respective controls (χ2=8.75, p=0.01) .And in third part of present study, disease status analysis for PAI-1 gene on the basis of DR stages (NPDR and PDR) was studied, which indicated statistically nonsignificance. The χ2 analysis values for DNR and NPDR and for DNR and PDR was (χ2=0.48, p>0.05)(χ2 =2.00, p>0.05) respectively, Conclusion: Our present study suggests that changes in genetic polymorphisms of ACE-gene and PAI-1 gene in DR, DNR and T2D Patients are risk factors, which may serve as useful prognostic markers.
ABSTRACT
The venom phosphodiesterase I (PDE-I, EC 3.1.4.1) is useful in the elucidation of the structure and nucleotide sequence of nucleic acids. In the present study, PDE-I was purified from Agistrodon bilineatus venom by preparative native-PAGE. A single protein band was observed in analytical native-PAGE. The enzyme also gave a single band in SDS-PAGE with a molecular mass of 140 kDa. The position of the band was not altered in the presence of β-mercaptoethanol, suggesting the protein did not contain subunits. The enzyme was free from 5’-nucleotidase and alkaline phosphatase activities. It showed a broad optimum pH range (9.0-11.0), whereas the optimum temperature was found to be 600C, with activity decreasing at >650C. Energy of activation (Ea) was calculated to be 0.31. The PDE-I was a glycoprotein having 14% of carbohydrate content. The Vmax, Km, Kcat and Ksp values of the enzyme were 3.85 μM/min/mg, 8.3 × 10-3 M, 23s-1 and 46.4 M-1 Min-1 respectively. Cysteine caused a non-competitive inhibition with a Ki 6.3 × 10−3 M (IC50 of 1.6 mM), whereas ADP caused a competitive inhibition having Ki 0.8 × 10−3 M (IC50 5.4 mM). Glutathione, o-phenanthroline, zinc and EDTA inhibited the enzyme activity, whereas Mg2+ slightly potentiated the activity. The enzyme hydrolyzed thymidine 5’-monophosphate p-nitro-phenyl ester most readily (10-fold), while 3’-5’-cAMP was least readily hydrolyzed substrate. The enzyme up to 4.0 mg/Kg i.p was not lethal in mice. It exhibited an anticoagulant effect, and increased the normal clotting time of normal citrated human plasma, whereas the crude venom showed strong coagulant effect. The above results showed that the A. bilineatus PDE-I was very similar to that isolated from other snake venoms. The purification procedure described here is simple, rapid and reproducible and may prove useful to isolate pure protein for investigation into the contribution of this enzyme to the biological activities of A. bilineatus venom and PDE-I insight, in general.