Open Access Original Research Article

Zymographic Detection of Aspartic Proteinase Activities in Porcine Ovarian Extracts

H. K. I. Perera, P. H. P. Fernando, S. B. P. Athauda

International Journal of Biochemistry Research & Review, Page 166-174
DOI: 10.9734/IJBCRR/2015/16236

Zymographic Detection of Aspartic Proteinase Activities in Porcine Ovarian Extracts

Aims: Tightly regulated proteolytic activity is essential in the mammalian ovary to maintain follicular and luteal functions. Studies conducted on ovarian aspartic proteinases (APs) are limited. Previously it has been noted that the AP activity increases towards the latter part of luteal phase. The aim of this study was to isolate AP from porcine ovarian extract and to identify whether multiple AP activities are found in the extracts.

Place and Duration of Study: Department of Biochemistry, between December 2009 and February 2012.

Methodology: Porcine ovaries (n= 100) were collected and ovarian extracts were prepared. APs were fractionated, using anion exchange chromatography at pH 8.5, gel permeation chromatography and affinity chromatography. AP activity (U/ml) of the fractions were measured in the presence and absence of Pepstatin A. AP specific activities (U/mg) were calculated after measuring total protein concentrations (mg/ml) of the fractions. Fractions were analyzed using polyacrylamide gel electrophoresis conducted under denaturing (SDS-PAGE) and native (PAGE) conditions. PAGE was followed by zymography.

Results: With anion exchange chromatography, AP was recovered during column washing as an unbound fraction (~67%) and during elution as a bound fraction (~33%). Bound AP was recovered around 0.23 M NaCl with 0-1 M NaCl gradient used during elution. Both AP fractions had an apparent molecular mass of 40 kDa. AP activity was completely inhibited in the presence of 1 µM Pepstatin. Specific activity of AP increased with fractionation from 1 in the ovarian extract to 747 and 511 U/mg with unbound and bound AP respectively. SDS-PAGE showed elimination of impurities with the progress of fractionation. PAGE and zymography showed the presence of at least three AP activities in porcine ovaries.

Conclusion: Proteinases fractionated using three chromatography procedures were APs. Results showed the presence of multiple AP activities in porcine ovary.

Open Access Original Research Article

Glycaemic Evaluation of Murraya koenigii in Alloxan-induced Diabetic Rabbits

T. Akande, S. T. Balogun, H. Abdullahi, T. O. Ogundeko, M. S. Ramyil

International Journal of Biochemistry Research & Review, Page 175-181
DOI: 10.9734/IJBCRR/2015/17529

Glycaemic Evaluation of Murraya koenigii in Alloxan-induced Diabetic Rabbits

Background: Study was aimed to evaluate the antidiabetic activity of Murraya koenigii, a traditional medicinal plant (Curry leaf) in normoglycemic and alloxan-induced diabetes rabbits.

Methods: Antidiabetic activity of aqueous extract of M. koenigii in 100, 200 300 mg/k doses was determined by estimating blood glucose before and at 1, 2, 4, 8, 24, and 72 hours post treatment intervals in treated rabbits.

Results: Aqueous extract of Murraya koenigii showed a dose dependent antidiabetic activity with maximum effect established at 300 mg/kg. The extract also exhibited a significant (p<0.05) dose- dependent hypoglycemic effect on normal and alloxan-induced diabetic rabbits.

Conclusion: Murraya koenigii causes a reduction in blood glucose. This hypoglycemic property supports-its usein folkloric medicine as an antidiabetic agent and thus suggests a place for it in nutritional therapy in the management of diabetes mellitus and thus as an oral hypoglycaemic agent.

Open Access Original Research Article

In vitro and In vivo Antioxidant Activity of the Total Dichloromethane-ethanol Extract of Morinda morindoides (Baker) Milne-redh. (ETDE) (Rubiaceae)

Boga Gogo Lucien, Bahi Calixte, Kouangbé Mani Adrien, Djaman Allico Joseph, N’Guessan Jean David

International Journal of Biochemistry Research & Review, Page 182-191
DOI: 10.9734/IJBCRR/2015/17382

Purpose: our work consisted in studying the antioxidant activity of the dichloromethane-ethanol total extract (Baker) milne-redh., a Rubiaceae with antihypertensive activity.

Methodology: The antioxidant activity was determined in vitro by the test of 2,2'-diphenyl-1-picrylhydrazyl (DPPH), the test of inhibition of lipid peroxidation by TBARS method and in vivo by determining activities of SOD, CAT and the concentration of NO in the organs of rats made hypertensive by adrenalin (ADR). In vivo, Rats were divided into 7 lots of 5 rats each. The control lot received daily orally distilled water for 14 days. The other lots received adrenalin (ADR) at dose of 1 mL/kg by intraperitoneal injection for 8 days. After installation of hypertension, lot MNT (Lot sick untreated or positive control) has not undergone any treatment. Rats of lots ETDE 500 and ETDE 1500, received orally ETDE at doses of 500 and 1500 mg/kg bw for 6 days. Rats of lots TEN (tenordate) 10 and TEN 20 received orally the tenordate at doses of 10 and 20 mg/kg pc for 6 days. After the 6 days of treatment, the rats are decapitated and their hearts, kidneys, livers andaortas were collected for the determination of oxidative stress parameters.

Results: Our results showed that ETDE, in vitro, reduces greatly the DPPH with a IC50 of 21.45±1.53% and inhibits 49.66±3.83% of lipid peroxidation. In vivo, ETDE reduced significantly and normalizes the activity of SOD, CAT and normalizes the concentration of NO of rats made hypertensive.

Conclusion: ETDE by its antioxidant action on free radicals may be beneficial in the treatment of high blood pressure.

Open Access Original Research Article

Gas Chromatography-Mass Spectrometry (GC-MS) Analysis of Ethanolic Extracts of Aerva lanata (L.)

Ramalingam Vidhya, Rajangam Udayakumar

International Journal of Biochemistry Research & Review, Page 192-203
DOI: 10.9734/IJBCRR/2015/17241

Aim: To determine the phytochemical constituents present in the different parts of Aerva lanata using Gas Chromatography – Mass Spectrometry (GC-MS).

Study Design: GC-MS analysis of bioactive compounds in different parts of A. lanata.

Place and Duration of Study: Post Graduate and Research Department of Biochemistry, Government Arts College (Autonomous), Kumbakonam and Department of Food Safety and Quality Testing, Indian Institute of Crop Processing Technology, Thanjavur, Tamilnadu, India, between May 2011 to June 2012.

Methodology: 15 g of powdered plant material of leaf, flower and root were soaked with 60 mL of 95% ethanol for 24 hrs. After 24 hrs, the extract was filtered and the filtrate was concentrated to        1 mL by bubbling nitrogen gas into the solution. 2 µL of ethanolic extracts of leaf, flower and root of A. lanata was used for GC-MS analysis.

Results: The GC-MS analyses showed that the presence of four different phytocompounds in ethanolic extract of leaf of A. lanata. The highest peak area of 74.73% for isophytol was identified in leaf of A. lanata. The ethanolic flower extract of A. lanata showed that the presence of twelve different phytocompounds. Flower extract contains the highest amount of phytocompound was 6, 9,12 –octadecatrienoic acid, phenyl methyl ester (z,z,z)- with the peak area of 25%. The root extract of A. lanata showed that the presence of eight different bioactive compounds. The root of A. lanata showed more quantity of lanost-9 (11)-en-12-one with the highest peak area of 45.11%.

Conculsion: The present study confirmed that the presence of active compounds in different parts of A. lanata. In future, the isolation of above mentioned bioactive compounds from the different part of A. lanata would be useful to find out the novel drugs.

Open Access Original Research Article

A Comparative Study of the Effect of Heating on Antioxidant Potential and Lipid Profile of Rats Fed with Two Vegetable Oils-sunflower Oil and Sesame Oil (Commonly Used in Culinary Preparation)

George Saramma, Chaturvedi Padmaja

International Journal of Biochemistry Research & Review, Page 204-211
DOI: 10.9734/IJBCRR/2015/17862

Aims: Vegetable oils are fats, which are extracted from plant materials especially from seeds, which may change its chemical composition when heated. Therefore the present study investigated the effect of heating of sunflower oil and sesame oil on the in-vivo antioxidant status and the lipid profile in Wistar albino rats.

Place and Duration of Study: Department of Biological Sciences, University of Botswana, Botswana. The study was conducted from January 2012 - December 2012.

Methodology: The animals were divided into five groups of six rats each and maintained as follows: Normal control received the normal diet and two positive controls treated with heated sunflower oil and sesame oil, two experimental groups-treated sunflower oil and sesame oil after frying the potato chips. At the end of the experimental period of 60 days the rats were sacrificed and the blood was collected for biochemical estimations. The weights of the animals were recorded every week.

Results: The results indicated that sesame oil is much better in keeping the antioxidant status and cause lesser lipid peroxidation as well as hepatotoxicity when compared to sunflower oil. Again the cardioprotective effect of sesame oil is higher than that of the sunflower oil.

Conclusion: So in conclusion, sesame oil showed a higher level of protective effect than that of sunflower oil and is a better choice for culinary purposes.