Purification and biochemical properties of carboxylesterase from fasciola gigantica

Carboxylesterase was purified from Fasciola gigantica through ammonium sulfate precipitation, chromatography on DEAE-Sepharose and gel filtration on a sephacryl S300. Three enzymes [El, EII and EIII] were separated. EII and EIII were purified to homogeneity. The molecular weight of EII and EIII enzyme were 66 and 50 KDa, respectively as detected by gel filtration and SDS-polyacrylamide gel electrophoresis. EII and EIII had Km 1.3 and 1.7 mM of p-nitrophenyl acetate. Affinity of esterase EII and EIII decreased as increasing carbon atom number of the substrates. Esterase EII and EIII had optimum temperature at 40 °C. Esterase EII and EIII had pH optima at pH 7.5 in phosphate buffer and pH 8.0 in Tris-HCl buffer, respectively. Studying effect of metal ions on esterase EII and EIII indicated that Li[+], Mn[++], Ba[++] and Mg[++] had activation effect on each isoenzyme. An activation effects could be detected with N- ethylmalimaide on EII and EIII]. The order of inhibition on EII was beta- mercaptoethanol > PMSF > DTNB > PCMB > iodoacetate. While the order of inhibition on EIII was beta-mercaptoethanol > iodoacetate > DTNB> PCM > PMSF

Possible mechanisms of acrylamide- induced hormonal imbalance in adult male rats

The current study aimed to investigate the possible mechanisms by which acrylamide affects vital endocrine glands in an experimental rat model. Thirty adult Sprague Dawley male rats were divided into three experimental groups, group [1] as control; group [2] where rats were orally administered acrylamide in a dose of 5 mg/kg b. wt. daily for 12 weeks; group [3] where rats were orally administered acrylamide in a dose of 15 mg/kg b. wt. daily for 12 weeks. Plasma was analyzed for malondialdehyde [MDA], total antioxidant capacity, total and free testosterone, T[3], T[4] and corticosterone levels. Histopathological examinations of testis, thyroid gland and adrenal gland were carried out. Our results indicate that acrylamide administration significantly increases plasma lipid peroxidation with concomitant decrease in total antioxidant capacity, total and free testosterone, T[3], T[4] and corticosterone levels. These results were documented by histopathological investigations of the testis, thyroid and adrenal tissues. We conclude that acrylamide causes hormonal disturbance in experimental animals via inducing oxidative and destruction stress on vital endocrine glands which presents a potential hazards to humans

Haptoglobin gene polymorphism in type 2 diabetic patients with and without nephropathy

The development and progression of diabetic microvascular complications including nephropathy are related to the degree of glycemic control and oxidative stress and may be influenced by genetic factors. The aim of the present study has been to investigate the association between haptoglobin [Hp] gene polymorphism and the occurrence of diabetic nephropathy in patients with type 2 diabetes mellitus and to find a possible link between haptoglobin phenotypes and inflammatory parameters; serum C - reactive protein [CRP], interleukin- 6 [IL - 6] and haptoglobin. The study included 60 normotensive type 2 diabetic patients of more than 5 years duration categorized in 3 equal groups according to urinary albumin excretion [UAE] into normo, micro and macroalbuminuric. In addition, 20 individuals of matched age and sex were selected to serve as a control group. Serum CRP, IL - 6 and haptoglobin concentrations were measured and haptoglobin phenotyping was conducted using polyacrylamide gel electrophoresis. The frequency of haptoglobin phenotype 1- 1 [Hp 1 - 1] in diabetic patients with normoalbuminuria was 7 of 20 [35%] as compared with diabetics with macroalbuminuria, 1 of 20 [5%] [p = 0.02] however, the frequency of Hp 2-2 was greater in diabetics with macroalbuminuria, 12 of 20 [60%] as compared with those with normoalbuminuria or controls, 5 of 20 [25%] [p= 0.03]. Patients with diabetic nephropathy [micro- or macroalbuminuria] had higher levels of serum CRP IL-6 and haptoglobin than those without nephropathy [normoalbuminuria]. Serum haptoglobin levels in type 2 diabetics were higher in Hp phenotype 2-2 as compared with Hp 1-1 however, serum CRP and IL - 6 levels did not differ significantly between haptoglobin phenotype groups. Moreover, there was significant positive correlations between urinary albumin excretion and serum levels of CRP, lL-6 and haptoglobin in diabetic patients. From this study, it could be concluded that haptoglobin phenotype 2-2 is considered as a major susceptibility gene for the development of nephropathy in type 2 diabetic patients. In addition, the significant association between inflammatory parameters and UAE indicates that inflammation may be a pathogenic mechanism of renal injury in type 2 diabetics. Moreover, serum interleukin - 6 and haptoglobin may be good prognostic factors for the development of nephropathy in the course of diabetes mellitus