Open Access Short communication

Effect of Carboplatin and Methotrexate on Lipid Levels in the Plasma Membrane of MCF-7 Cells and their Association with Cell Motility

J. Borjac, R. Dannaoui, H. Saab, M. Dakdouki, M. El-Sibai, J. Usta

International Journal of Biochemistry Research & Review, Page 1-12
DOI: 10.9734/IJBCRR/2015/20709

Aim: We examine in this study the effect of two antitumor drugs metothrexate (MTX) and carboplatin (CPT) on the phospholipid content of plasma membrane of MCF-7 breast cancer cells. MTX is a folate analog that inhibit dihydrofolate reductase. CPT is a platinum-containing antineoplastic drug that inter-chelates DNA and inhibits replication.

Methods: MCF-7 breast cancer cells were treated with different concentrations of CPT or MTX.  Plasma membranes were purified and their protein contents were measured with and without drug treatment. We extracted the lipids from plasma membranes of drug-treated and control MCF-7 cells, quantitated, and separated by HPLC using a C18 reverse-phase column.

The ability of both drugs to affect cell movement was studied using a motility assay.

Results: MTX induced 50% cell death at concentrations ranging from 50 to 100 mM. No further decrease in viability was seen above this concentration even at 1 mM. CPT induced 50% cell death at 5 mM. It also showed a range concentration that gives a 50% cell death after 24 hrs of incubation. Protein levels in plasma membranes of treated cells doubled compared to control. Lipid levels in plasma membranes decrease insignificantly after drug treatment. No changes in separation pattern of lipid extracted from membranes were seen after CPT treatment compared to control; however, MTX treatment showed to a single change in elution pattern in peak at 4.36 min.

Both drugs had little effect on cell motility causing a decrease of 13.3% and 17.4% with CPT and MTX respectively compared to control.

Conclusions: Our results show that both drugs did not affect the amount of lipids but lead to doubling in the protein concentration in the plasma membranes of MCF-7 breast cancer cells.  These drugs did not lead to a significant decrease in cell motility compared to control.

Open Access Original Research Article

Evaluation of the Effect of Starvation on Blood Glucose Level and Body Weight of Wistar Albino Rats

Okoro Onyinyechi Ruth, Eze Chinemelum Adaora, N. Ogugua Victor, Onyia Chukwuebuka Felix, C. Eluu Stanley, Ejikeugwu Chika

International Journal of Biochemistry Research & Review, Page 1-8
DOI: 10.9734/IJBCRR/2015/20721

Aims: The aim of this study was to determine the effect of food deprivation on the body weight and blood glucose level of Wistar albino rats.

Methodology: Blood samples were collected from the rats through ocular puncture at intervals and were used for the analysis of blood glucose level. Standard procedures was used for the measurements, and blood glucose level was evaluated using the ONE TOUCH () blood glucose monitoring system/meter.

Results: According to the duration of study for both the control and experimental animals, the body weight and glucose levels of the rats were taken before and after the starvation. There was no significant increase (p>0.05) in the body weights of the test animals compared with the body weights of the animals in control group at 0 hour and 6 hours  of the experiment. However, there was a significant increase (p<0.05) in the body weights of treated rats after 12 hours when compared with the control group. After 24 hours treatment, group 2 animals (animals starved of rat feed and water) had significant increase (p<0.05) in their mean body weight when compared with control. There was significant increase (p<0.05) in the rats’ body weights of group 4 treated animals (animals starved and received fruits only) compared with the control group after a period of 6 hours.  There was also a significant increase (p<0.05) in the body weights of treated group 4 rats (animals starved and received fruits only) after 12 hours when compared with the control group. For 48 hours treatment, no significant difference (p>0.05) in body weight was observed. The glucose concentration increased significantly (p<0.05) in group 3 test animals administered water after starvation compared with the control animals at the 0 hour duration of the study. There was significant decrease (p<0.05) in the glucose concentration of animals (group 4) fed fruit after starvation compared with the animals (group 3) administered water after starvation at 0 hour of the experiment. However, the glucose concentrations of the animals in group 2 (starved of feed and water) and group 4 (starved + fruit) were not significant (p>0.05) compared with the control. The blood glucose concentrations of rats in groups 2 and 3 increased significantly (p<0.05) when compared with the control (group 1) within the duration of 6 hours.

Conclusion: Conclusively, this study shows that starvation has significant effect on the blood glucose level and body weight of Wistar albino rats. Such significant data (as obtainable in this study) could be extrapolated to humans with a view to unraveling the untoward consequences of starvation on the human body system.

Open Access Original Research Article

Detection of High Risk Human Papilloma Virus (HPV) Genotypes 16/18 in Oral Lesions of Tobacco Chewers in Pakistan

Zile Rubab, Saeeda Baig, Mohammad Haris Lucky

International Journal of Biochemistry Research & Review, Page 1-7
DOI: 10.9734/IJBCRR/2015/20761

Background: The Human Papilloma Virus (HPV) has evolved as a new culprit of malignant and pre malignant oral lesions. The objective of this study was to find out the frequency of HPV and its high risk genotypes in different lesions of oral cavity of tobacco chewers.

Methods: From 492 subjects (421 males and 71 females), 20 ml of oral rinse sample was collected after obtaining an informed consent. Normal subjects with no chewing habits (250) including 135 males and 115 females were also taken from same setting. Gentle brushings over the lesions with the help of dental floss brush was done which was left in the oral rinse and stored at 4°C until DNA extraction. DNA was extracted and PCR was performed using HPV consensus primers Gp5+/Gp6+ and HPV 16, 18 specific primers for genotyping. Categorical data was calculated as frequencies and percentages.

Results: Oral pre-malignant lesions were present in 421 (86%) males and 71(14%) females having leukoplakia (173, 35%), erythroplakia (60, 12%), submucous fibrosis (192, 39%) and L/E (67, 14%). Total number of HPV positives were 128 (26%), having HPV 16 (13%) and HPV 18 (11%) whereas, 76% had other genotypes. Among Submucous fibrosis 82(46%) were HPV positive. Out of total 128 HPV patients 92% were males and 8% were females. All controls were found to be HPV negative.

Conclusion: Frequency of HPV was found high (26%) in oral lesions with HPV16/18 as 13% and 11% respectively. The patients with submucous fibrosis are at greater risk of having HPV. Other HPV genotypes causing premalignant lesions require further investigation.

Open Access Original Research Article

Inhibitory Activity of Ethanolic Extract of Horseradish (Moringa oleifera) (Lam) on Oxidative Stress Parameters in Some Organs of Dexamethazone Induced Hypertensive Wistar Rats

O. A. Oseni, A. S. K. Idowu, T. O. Obaparusi

International Journal of Biochemistry Research & Review, Page 1-10
DOI: 10.9734/IJBCRR/2015/18455

The use of herbal remedies in various local communities throughout the world to assuage the scourge of indigenous diseases has gained tremendous popularity. One of the many plants species of herbal remedies is Moringa oleifera (horseradish) which has been widely used in West Africa and Nigeria in particular for treating numerous human ailments such as malnutrition, cardiovascular, hepatotocicity and many others. Regardless of its wide use by communities around the world, there is inadequate scientific information available on the actual pharmacological effects of ethanolic extract of Moringa oleifera leaves on the heart and kidney dysfunction.  Hence this work aimed at determining the inhibitory activity of ethanolic extract of horseradish on oxidative stress in the heart and kidney of dexamethazone induced hypertensive wistar albino rats. The air-dried leaves of horseradish were pulverized and crude ethanolic extract was prepared. The animals were grouped into four groups of five animals each. Group A animals served as control and fed with water and feed while Groups B and C animals were administered with 0.5 mL of 40% w/v ethanolic extract of Moringa oleifera leaf and 0.5 ml of 1.67 mg/kg body weight dexamethasone respectively. Group D animals were treated with 0.5 ml of 1.67 mg/kg body weight dexamethasone and 0.5 mL of 40% ethanolic extract of Moringa oleifera leaf intermittently for 21 days. The results obtained from the study showed that the ethanolic extract of Moringa oleifera leaf possessed the ability to inhibit and reverse the dexamethasone mediated tissues oxidative damage of both organs as seen in cholesterol, reduced glutathione and malondialdehyde concentrations. The same protective trend was also observed in activities of alkaline phosphatase, aspartate transaminase, alanine transaminase and catalase enzymesin both organs of the hypertensive rats. Hence, ethanolic extract of Moringa oleifera leaf reduced the extent of antioxidant loss and restoration of organ dysfunction caused by dexamethasone in the rats.

Open Access Original Research Article

HPLC-PDA-ESI-MS Identification of Polyphenolic Phytochemicals in Different Market Classes of Common Beans (Phaseolus vulgaris L.)

V. Nyau, S. Prakash, J. Rodrigues, J. Farrant

International Journal of Biochemistry Research & Review, Page 1-11
DOI: 10.9734/IJBCRR/2015/21608

Aim: The aim of the study was to screen the market classes of common beans popularly grown in Zambia for their polyphenolic phytochemical profiles.

Methodology: Three market classes of common bean (red, grey mottled and brown) were investigated for their polyphenolic phytochemical profiles using the High Performance Liquid Chromatography-Photo Diode Array-Electrospray Ionization-Mass Spectrometry (HPLC-PDA-ESI-MS).

Results: Various phenolic compounds were identified in the Zambian market classes of common beans. Quinic acid, a syringic acid derivative, ferulic acid derivatives, medioresinol, p-coumaric acid, catechin, gallic acid and ferulic acid were identified in all the market classes investigated. However, the isomers of ferulic acid derivatives observed were 4 in red, 3 in brown and 1 in grey mottled beans. Epicatechin was only identified in the red and grey mottled beans. A compound with the molecular ion at m/z 567, tentatively identified as a flavonone derivative was only observed in red beans. Catechin glucoside was only identified in grey mottled and brown beans. Compounds tentatively identified as kaempferol glucoside and carnosol were only observed in brown beans and not in the other market classes.

Conclusion: The study has shown that the market classes of common beans investigated contain various polyphenolic compounds that may be useful as nutraceuticals. The study assumes that the diversity in the phenolic phytochemical profile is an excellent opportunity for genetic improvement in the nutraceutical attributes of these common bean market classes by crop breeders.

Open Access Original Research Article

Purification and Characterization of Endoglucanase from Sorghum (S. bi-color) and Millet (Pennisetum typhoides and Digitaria exilis) Malts

D. I. Ukairo, H. C. Nzelibe, C. I. Iheme, E. C. Udenze, C. P. Ihedimbu, A. U. Ezirim

International Journal of Biochemistry Research & Review, Page 1-11
DOI: 10.9734/IJBCRR/2015/20593

Endoglucanase (EC3.2.1.4) from sorghum (S. bi-color) and millet (Pennisetum typhoides & Digitaria exilis) malts were purified to homogeneity through the methods of ammonium sulphate precipitation and gel filtration. Molecular mass of 35 KDa and 41 KDa were determined by SDS-PAGE. The purified enzymes catalyzed the hydrolysis of carboxy-methylcellulose with optimum activity at pH of 4.8, 5.0, 6.0, and temperature of 60ºC, 60ºC and 70ºC for Digitaria exilis, S. bi-color and Pennisetum typhoides respectively. More than 90% activity was retained in S. bi-color and Pennisetum typhoides and 73% activity in Digitaria exilis after 1.0 hour pre-incubation at 60ºC. Km values of 0.11, 0.09, 0.20 mM and Vmax 17.53, 15.0 and 11.10 U/mg/min were obtained for S. bi-color, Pennisetum typhoides and Digitaria exilis respectively. Co2+ inhibited endoglucanase activity whereas Ca2+, Ba2+, and Zn2+ enhanced enzyme activity. The enzyme was inactivated by glucose, a major end product of cellulose hydrolysis. Results indicate that endoglucanase of S. bi-color and Pennisetum typhoides are more suitable for malting and a blend of the two will produce high quality malt.