Evaluation of Mango Seed Kernel Methanolic Extract on Metalloproteases in Carpet Viper (Echisocellatus) Venom: An in Vitro Experiment

Background: The global incidence of snakebite has become a major concern to the community. This study aimed to evaluate the effect of mango seed kernel methanol extract on metalloproteases in Carpet Viper (Echis ocellatus) venom. Methods: Mango seed kernel methanolic extract was evaluated in vitro for its anti-venom activity and inhibition of metalloproteases of Carpet Viper's (Echis ocellatus) venom. Metalloprotease portion was partially purified from the venom of E. ocellatus with a yield of 71%, a purification fold of 2.63 and a specific activity of 19.00 µmol/min/mg protein. Results: The enzyme appeared as a band on SDS-PAGE with a molecular weight of 23 kDa. The kinetic properties of the enzyme showed a Km of 0.31 mg mL-1 and a Vmax of 9.09 µmol min-1. When the enzyme was incubated with the extract, kinetic studies revealed a mixed non-competitive pattern of inhibition with Km values of 0.56 and 1.11 mg mL-1 and Vmaxvalues of 6.67 and 4.17-µmol min-1 for 5% and 20% inhibitor concentrations, respectively. An estimated Ki value of 0.168 mg mL-1 was obtained from a secondary plot demonstrating that the extract had a high affinity for the partially purified enzyme; thus, could serve as an effective inhibitor. Conclusion: Methanol extract of mango seed kernel has a high affinity for the partially purified enzyme, and it might provide an inexpensive and readily available alternative to sheep serum in the management of snakebite envenomation. Therefore, further in vivo studies are necessary to assess its effectiveness and safety

Validation of the antidiabetic effects of Vernonia amygdalina delile leaf fractions in fortified diet-fed streptozotocin-treated rat model of type-2 diabetes

Background: Vernonia amygdalina (VA) is used in the traditional management of diabetes in Nigeria. Previous scientific verification of VA is on Type-1 diabetes model, in spite of the continuous increase in Type-2 diabetes (T2D) among adults. This study aimed to validate the antidiabetic effects of VA leaf fraction (VALF) in a unique T2D rat model. Materials and Methods: Methanol crude extract of VA leaf was fractionated with solvents of increasing order of polarity (n-hexane, chloroform, ethyl-acetate, n- butanol and water). The antidiabetic activities of the fractions were evaluated in vivo in T2D model rats. Albino Wistar rats were induced with T2D and treated with the VALF. Several T2D-related parameters were measured. Results: T2D rats showed significant increase in serum levels of fasting blood glucose (FBG), liver and kidney biomarkers. At 28-day post-oral treatment with the VALF, FBG levels were significantly (P < 0.05) reduced (n- hexane [29.3%], chloroform [66.7%], ethyl acetate [36.2%], n- butanol [45.59%] and aqueous [39.3%]). The glucose tolerance ability was significantly improved in the chloroform fraction (Vernonia amygdalina chloroform fraction [VAc])-treated groups compared to the other fractions-treated group and diabetic control group. Furthermore, the VAc was found to be most effective as it ameliorates most of the alterations caused in the studied parameters in diabetic rats when compared with n- hexane, ethyl acetate, n- butanol and aqueous fractions. Conclusion: The study validates the anti-diabetic effects of VALF in fortified diet-fed streptozotocin-treated rat model of T2D, and suggests that the VAc is a potential candidate for development of a more effective drug for the management of T2D

Appropriate Insulin Level in Selecting Fortified Diet-Fed, Streptozotocin-Treated Rat Model of Type 2 Diabetes for Anti-Diabetic Studies.

Pathophysiological investigation of disease in a suitable animal model is a classical approach towards development of a credible therapeutic strategy. This study examined appropriate insulin level in selecting animal model for type 2 diabetes (T2D) studies.Albino Wistar rats (150-200g) were divided into two groups fed with commercially available normal-diet-feed (NDF) and water or fortified diet feed (FDF) (10g NDF per gram of margarine) with 20% fructose solution as drinking water. After 6 weeks of dietary regimen both groups were divided into 5 sub-groups and injected intraperitoneally with a graded dose of streptozotocin (STZ) (0, 25, 35, 45 & 55mg/kg bw.).The result showed that the FDF-fed rats increased significantly in body weight, basal serum insulin, total cholesterol, triglycerides and blood glucose levels as compared to NDF-fed rats. Ten days post STZ induction, the groups treated with STZ (45 & 55 mg/kg) developed frank hyperglycaemia with 300 mg/dL) throughout the 6weeks post diabetes confirmation. These FDF35 rats were sensitive to glibenclamide, metformin and pioglitazone in lowering hyperglycaemia, hypertriglyceridemia and hypercholesterolemia.The hyperglycaemia stability of the FDF35 rats (85.5% insulin) together with their sensitivity to 3 different hypoglycaemic drugs strongly suggests their suitability as a non-genetic model of T2D. Hence the study shows that circulating serum insulin ≥ 85.8% with overt hyperglycaemia may be utilized as the benchmark in selecting rat models for T2D studies

Tannery Effluent Treatment by Yeast Species Isolates from Watermelon

The quest for an effective alternative means for effluent treatment is a major concern of the modern-day scientist. Fungi have been attracting a growing interest for the biological treatment of industrial wastewater. In this study, Saccharomycescerevisiae and Torulasporadelbrueckii were isolated from spoiled watermelon and inoculated into different concentrations of effluent. The inoculants were incubated for 21-days to monitor the performance of the isolates by measurement of biochemical oxygen demand (BOD), chemical oxygen demand (COD), nitrates, conductivity, phosphates, sulphates and turbidity. The results showed that Saccharomycescerevisiae had the highest percentage decrease of 98.1%, 83.0%, 60.7%, 60.5%, and 54.2% for turbidity, sulphates, BOD, phosphates and COD, respectively, of the tannery effluent. Torulasporadelbrueckii showed the highest percentage decrease of 92.9%, 90.6%, and 61.9% for sulphates, COD, and phosphates, respectively, while the syndicate showed the highest percentage reduction of 87.4% and 70.2% for nitrate and total dissolve solid (TDS), respectively. The least percentage decrease was displayed by syndicate organisms at 51.2%, 48.1% and 40.3% for BOD, COD and conductivity, respectively. The study revealed that Saccharomycescerevisiae and Torulasporadelbrueckii could be used in the biological treatment of tannery-effluent. Hence, it was concluded that the use of these organisms could contribute to minimizing the adverse environmental risks and health-hazards associated with the disposal of untreated tannery-effluents

Oral Glucose Tolerance Test.

<p>Data are presented as mean ± SD of 6–8 animals per group. <b>NDF:</b> Group fed with Normal Diet Feed + water and treated with normal saline as control <b>NDF</b>_<b>25</b><b>:</b> Group fed with Normal Diet Feed + Water and treated with 25mg/kg bw, STZ. <b>NDF</b>_<b>35</b><b>:</b> Group fed with Normal Diet Feed + Water and treated with 35mg/kg bw, STZ. <b>NDF</b>_<b>45</b><b>:</b> Group fed with Normal Diet Feed + Water and treated with 45mg/kg bw, STZ <b>NDF</b>_<b>55</b><b>:</b> Group fed with Normal Diet Feed + Water and treated with 55mg/kg bw, STZ. <b>FDF:</b> Group fed with Fortified Diet Feed + 20% fructose and treated with Normal saline <b>FDF</b>_<b>25</b><b>:</b> Group fed with Fortified Diet Feed + 20% fructose and treated with 25mg/kg bw STZ <b>FDF</b>_<b>35</b><b>:</b> Group fed with Fortified Diet Feed + 20% fructose and treated with 35mg/kg bw STZ <b>FDF</b>_<b>45</b><b>:</b> Group fed with Fortified Diet Feed + 20% fructose and treated with 45mg/kg bw STZ <b>FDF</b>_<b>55</b><b>:</b> Group fed with Fortified Diet Feed + 20% fructose and treated with 55mg/kg bw STZ.</p

Effect of STZ Dose on Percentage Change in Serum Insulin, C-Peptide and Blood Glucose levels in all the groups with respect to the NDF control group.

<p>Effect of STZ Dose on Percentage Change in Serum Insulin, C-Peptide and Blood Glucose levels in all the groups with respect to the NDF control group.</p

Non-Fasting Blood Glucose Curve in FDF₃₅ Rat Group from Post STZ Induction to the End of the Experiment.

<p>Data are presented as mean ± SD of 6 animals.</p

Effect of Pioglitazone, Glibenclamide and Metformin on Blood Glucose, Total Cholesterol and Triglyceride in FDF-Fed, STZ Treated Rat Model of Type 2 Diabetes.

<p>Data are presented as mean ± SD of 6 animals per group. <b>TC:</b> total cholesterol, <b>TG:</b> triglyceride, <b>FBS:</b> fasting blood sugar. <b>FDF</b>_<b>35</b><b>:</b> Group fed with Normal Diet Feed + Water and treated with 35mg/kg bw, STZ.“a, b, c, d”<i>P</i> ≤ 0.05versus“FDF₃₅ + Vehicle”. Conversion factor: Insulin (1U/l = 7.174 pmol/l).</p

Mean Fluid Intake and Feed Intake of each Animal in the Groups per Day during the Experimental Period.

<p>Data are presented as mean ± SD of 6–8 animals per group. Bars with different superscript indicate significant difference (P<0.05). <b>NDF:</b> Group fed with Normal Diet Feed + water and treated with normal saline as control <b>NDF</b>_<b>25</b><b>:</b> Group fed with Normal Diet Feed + Water and treated with 25mg/kg bw, STZ. <b>NDF</b>_<b>35</b><b>:</b> Group fed with Normal Diet Feed + Water and treated with 35mg/kg bw, STZ. <b>NDF</b>_<b>45</b><b>:</b> Group fed with Normal Diet Feed + Water and treated with 45mg/kg bw, STZ <b>NDF</b>_<b>55</b><b>:</b> Group fed with Normal Diet Feed + Water and treated with 55mg/kg bw, STZ. <b>FDF:</b> Group fed with Fortified Diet Feed + 20% fructose and treated with Normal saline <b>FDF</b>_<b>25</b><b>:</b> Group fed with Fortified Diet Feed + 20% fructose and treated with 25mg/kg bw STZ <b>FDF</b>_<b>35</b><b>:</b> Group fed with Fortified Diet Feed + 20% fructose and treated with 35mg/kg bw STZ <b>FDF</b>_<b>45</b><b>:</b> Group fed with Fortified Diet Feed + 20% fructose and treated with 45mg/kg bw STZ <b>FDF</b>_<b>55</b><b>:</b> Group fed with Fortified Diet Feed + 20% fructose and treated with 55mg/kg bw STZ.</p