Open Access Original Research Article

Temperature Induced Conformational Entropy of α-Amylase with and without Additive

Ikechukwu I. Udema

International Journal of Biochemistry Research & Review, Page 1-12
DOI: 10.9734/IJBCRR/2017/31097

Aims: 1) To formulate models based on defined principle for the application of Fitter’s model and 2) ultimately show that there are changes in the radius of an enzyme in solution and consequently conformational entropy change with temperature before and during catalytic activity.

Study Design: Theoretical and experimental.

Place and Duration of Study: Department of Chemistry and Biochemistry, Research Division, Ude International Concepts LTD (862217), B. B. Agbor, Delta State, Nigeria; Owa Alizomor Secondary School, Owa Alizomor, Ika North East, Delta State, Nigeria. The research was conducted between June and December, 2016.

Methodology: Bernfeld method of enzyme assay was used. Assays were carried out on diluted crude human salivary alpha amylase (HSaA). Data from assay were plotted in two ways viz: Velocity (v) vs thermodynamic temperature (T) and 1/v vs T. Proportionality constants were determined from slope and intercept of the linear regression. These were used to determine desired parameters.

Results: The results showed that, hydrodynamic radius, rp0 (or positive Dr) was higher at higher temperatures and in the absence of calcium salt. Without calcium chloride but with raw starch and gelatinized starch, Dr values were in the range of 0.81 Å-2.30 Å and 4.5 Å-8.45 Å respectively. With calcium chloride and gelatinized starch at higher temperatures, Dr ranged from 3.71 Å-4.80 Å. Increasing values of rp0 corresponded with increasing values of conformational entropy change, DSconf.

Conclusion: The derived models were theoretically/conceptually and experimentally compatible given that the hydrodynamic radius, rp0 (or positive Dr) increased with the increase in temperature whether or not the enzyme showed increasing activity. The activity of the enzyme decreases when physiological limit of DSconf is exceeded. The values of Dr were higher at higher temperatures in the presence and absence of calcium chloride.

Open Access Original Research Article

Hepatic Involvement with Elevated Liver Enzymes in Nepalese Subjects with Type 2 Diabetes Mellitus

Nirajan Shrestha, Nirmal Prasad Bhatt, Puja Neopane, Sudimna Dahal, Prashant Regmi, Madhav Khanal, Rojeet Shrestha

International Journal of Biochemistry Research & Review, Page 1-8
DOI: 10.9734/IJBCRR/2017/31935

Background: Diabetes mellitus (DM) is characterized by a broad disturbance in metabolism. Since the liver plays a vital role in the regulation of metabolism, there is evidence of liver dysfunction in DM. There is a high prevalence of non-alcoholic fatty liver disease among diabetic patients in western countries. However, there is a paucity of data showing of such association in Nepalese patients with DM. Therefore, the present study was carried out for a better understanding of alteration of liver function in patients with type 2 DM (T2DM).

Methods: A total of 105 patients (age between 30-74 years, male/female ratio=0.6), newly diagnosed as T2DM, without the history of liver disease and other chronic diseases, were recruited from out-patient department of Nepal Medical College Teaching Hospital along with 58 age and sex matched healthy controls (age between 30-71 years, male/female ratio=0.7). After basic anthropometric measurements, fasting venous blood was collected and subjected to analysis for liver enzymes.

Results: We observed marginal, yet statistically significant increase of serum alanine transferase (26.6±0.84 (diabetic) vs 20.0±0.69 (control), p=0.003) and γ-glutamyl transferase (40.4±1.51(diabetic) vs 21.2±1.1(control), p<0.001) in the diabetic patients compared to healthy controls.

Conclusion: The liver enzymes were marginally elevated in T2DM compared to controls. This increment may be associated with early pathological changes in the liver. Hence, regular monitoring of liver function is highly beneficial to prevent advance liver injury in T2DM.

Open Access Original Research Article

Circulating Survivin and TIMP-1 in Hepatitis C Virus Associated Liver Fibrosis

Bakheet E. M. Elsadek, Ahmed A. Abdel Ghany, Soad M. Abdel Ghany, Shamardan E. S. Bazeed, Mohamed A. Abdel Aziz, Salama R. Abdel Raheim, Abdullatif A. Ahmed

International Journal of Biochemistry Research & Review, Page 1-13
DOI: 10.9734/IJBCRR/2017/31609

Aims: The present study was conducted to assess the clinical utility of survivin, an anti-apoptotic protein, and the profibrogenic tissue inhibitor metalloproteinase-1 (TIMP-1) as non-invasive biomarkers for the discrimination between stages of HCV-associated liver fibrosis that are largely asymptomatic.

Methodology: Circulating survivin and TIMP-1 levels as well as their corresponding proteins and genes expressions were, respectively, assessed by ELISA, Western blot and RT-PCR in 100 HCV-infected patients at different stages of liver fibrosis in comparison to healthy controls.

Results: With the progression of hepatic fibrosis, each of circulating survivin and TIMP-1 as well as their ratio (survivin/TIMP-1) showed a stepwise increase and exhibited significant positive correlations with the fibrotic stage (r=0.98 & p=0.002, r=0.95 & p=0.01, and r=0.93 & p=0.013 respectively). A gradual increase in both survivin and TIMP-1 genes expression was also observed.

Conclusion: In conclusion, survivin, TIMP-1, and their ratio could represent promising biomarkers for prediction and discrimination of different stages of HCV-associated liver fibrosis. The higher sensitivity and specificity of survivin may provide new insights into its possible use as a target for the antifibrotic drugs. Further studies with validated tests on a large scale prospective clinical trial are required to ascertain these results.

Open Access Original Research Article

Formulation, Nutritional Evaluation and Consumer Acceptability of High Energy and Protein Dense Complimentary Foods from Millet and Soybeans

N. M. Banki, N. Danbaba

International Journal of Biochemistry Research & Review, Page 1-12
DOI: 10.9734/IJBCRR/2017/30581

This study was designed to formulate complementary foods from millet and soybean blends with the aim to provide high energy and proteineous food for vulnerable populations. Single screw extruder was used for the extrusion of the millet-soybeans composite blends and response surface methodology in a central composite rotatable design were adopted for the design of the process variables that could produce optimum complementary foods with high protein, calorie values and physical attributes. Results indicated that during extrusion, the colour of the extrudates varied between 17.67 and 54.81. The expansion index of the formulations ranges between 121.10 to 149.84. However, the results of proximate composition reported that moisture content ranges from 0.18% to 0.35%. The protein content increased from 23.81±1.21 to 28.87±0.16 suggesting proportional increase in protein when millet was blended with soybeans. The fat content varied between 0.08% and 0.10%. With respect to the carbohydrate and calorie values, the highest values were 89.56% and 494.53 kcal/100 g which was found in sample extruded at temperature of 100°C with 25% soybeans composition and 8% feed moisture content while the least values were observed in samples extruded at 140°C with 25% soybeans composition and 8% feed moisture content for carbohydrates (81.06%) and for calories (432 kcal). The result of pasting characteristics showed effect on the variation observed in the peak viscosity of different extruded products. Peak viscosity for the extrudates varied from 113.83 to 228.68. The consumer’s acceptability of the extrudates after reconstituting into gruel with warm water, milk and sugar showed that all the samples were generally acceptable.

Open Access Original Research Article

Synergistic Effect of Ethanol Extracts of Moringa oleifera and Pleurotus ostreatus on Liver Enzymes and Some Renal Functions of Alloxan-induced Diabetic Wistar Albino Rats

T. A. Nnadiukwu, C. C. Monago, L. C. Chuku

International Journal of Biochemistry Research & Review, Page 1-11
DOI: 10.9734/IJBCRR/2017/29014

Objective: To evaluate the effect of the combined ethanol extracts of Moringa oleifera and Pleurotus ostreatus on the liver and some renal function of alloxan-induced diabetic rats by checking the concentrations of the liver enzymes (AST, ALT & ALP), total bilirubin, total protein, albumin, creatinine, urea & uric acid.

Study Design: Animal experimental study.

Place of Study: Department of Biochemistry, Faculty of Science University of Port Harcourt P.M.B 5323 Port Harcourt Nigeria.

Year of Analysis: 2015.

Methods: The animals were induced diabetes mellitus type 1 with alloxan at 120 mg/kg body weight intraperitoneally. The combined extract was administered to the diabetic wistar albino rats at a different combination percentage of 60% (1,800 mg/kg) Moringa oleifera and 40% (600 mg/kg) Pleurotus ostreatus and 40% (1,200 mg/kg) Moringa oleifera and 60% (900 mg/kg) Pleurotus ostreatus respectively. The rest of the diabetic rats were treated with 100% (3,000 mg/kg) Moringa oleifera, 100% (1,500 mg/kg) Pleurotus ostreatus while 7.1mg/kg of metformin was used as the standard drug. The effect of the combined treatment of the extracts of Moringa oleifera and Pleurotus ostreatus on the liver and renal function of the diabetic animals were monitored by measuring the concentrations of the liver enzymes (alkaline phosphatase, alanine transaminase and aspartate transaminase), total bilirubin, total protein, albumin, creatinine, urea and uric acid.

Results: The combined ethanol leaf extracts of Moringa oleifera and Pleurotus ostreatus significantly (p<0.05) lowered the ALT, AST, ALP, total bilirubin and uric acid and significantly increased the total protein concentration in all the diabetic rats been treated when compared with the diabetic (disease) control rats that were not treated. It was also observed that there was no significant (p<0.05) difference in the concentration of albumin, creatinine and urea during the period of treatment.

Conclusion: This study concluded that the combined ethanol extracts of Moringa oleifera and Pleurotus ostreatus produced a significant reduction of elevated liver enzymes and are capable of controlling/regulating impaired kidney functions.