Open Access Original Research Article

Differential Regulation of E-Cadherin Expression by the Soluble Ectodomain and Intracellular Domain of Jagged1

Craig Delury, Claire Tinker, Samantha Rivers, Matthew Hodges, Susan Broughton, Edward Parkin

International Journal of Biochemistry Research & Review, Page 278-290
DOI: 10.9734/IJBCRR/2013/3847

Aberrant Jagged1-mediated Notch activation is linked to cancer and induces epithelial-to-mesenchymal transition through the repression of E-cadherin transcription. All three proteins are subject to sequential proteolytic events referred to as regulated intramembrane proteolysis. This process releases soluble protein ectodomains from the cell and, concomitantly, generates intracellular domains capable of nuclear translocation and transcriptional regulation.

Aim: To determine the cognate roles of the Jagged1 ectodomain and intracellular domain fragments in the regulation of E-cadherin expression.

Methodology: Human embryonic kidney cells were stably transfected with coding DNA constructs analogous to full-length Jagged1, the soluble Jagged1 ectodomain, or the intracellular domain fragment of the protein. Correct construct expression and processing were confirmed by immunoblot analysis of transfectant cell lysates and conditioned culture medium. The effects of the various Jagged1 constructs on endogenous E-cadherin expression and processing were subsequently monitored by immunoblot and RT-qPCR analyses.

Results: Both full-length Jagged1 and the soluble Jagged1 ectodomain construct down-regulated E-cadherin expression at the protein and RNA level. In contrast, the Jagged1 intracellular domain fragment construct enhanced E-cadherin expression but only at the RNA level.

Conclusion: The soluble Jagged1 ectodomain is sufficient for the down-regulation of E-cadherin expression whereas the intracellular domain of the protein does not exhibit such an effect and actually increases E-cadherin RNA expression. These results raise the interesting possibility of E-cadherin regulation in cells distal to the site of soluble Jagged1 ligand generation.

Open Access Original Research Article

Cardiac Tolerance Study of Morinda morindoides (Baker) Milne-Redhead (Rubiaceae) Extract in Rabbit

Irie Otis Tra-Bi, Bernard Nazaire Djyh, Amadou Founzegue Coulibaly, Allico Joseph Djaman, Jean David N’guessan

International Journal of Biochemistry Research & Review, Page 291-302
DOI: 10.9734/IJBCRR/2013/4251

Purpose: This study was conducted  to  assess  the  effect  of  the ethyl acetate fraction (AcEF1) obtained from the 96% extract of Morinda morindoides extract on cardiac tissue integrity and function in rabbits.

Methodology: The rabbits were divided in five groups of 6 rabbits each. Groups 2,3,4 and 5  received intraperitoneally, twice a week during four weeks, the ethyl acetate fraction at doses going from 25 to 100 μg/kg body weight while Group 1, received 1 mL of Mac Ewen fluid. Blood sampling was carried out to evaluate aspartate aminotransferase (AST), alanine aminotransferase (ALT), Creatine phosphokinase (CPK), Lactate dehydrogenase  (LDH) activities and level of calcium, sodium, potassium, magnesium and chloride in serum. The eighth week of the experiments, histopathological studies were also conducted on rabbits heart.

Results: Analysis of the serum markers showed slight increases in AST, ALT, CPK activities as well as sodium, calcium and potassium concentrations (p<0.05).  LDH activity, magnesium and chloride concentrations were unchanged compared to their initial values.

Histopathological studies had not revealed damages in the structure of the heart.

Conclusion: The ethyl acetate fraction did not exert any noxious effect on cardiac tissues and should be no toxic for cardiac tissues.

Open Access Original Research Article

Effect of Para Substituted Anilines Chemical Structure on Methane Biosynthesis by the Methanogens

K. Kayembe, L. Basosila, K. Kalala, A. K. Kitambala, C. P. Sikulisimwa, P. T. Mpiana

International Journal of Biochemistry Research & Review, Page 303-314
DOI: 10.9734/IJBCRR/2013/2458

Aims: The present paper aims to study the effect of aromatic structure on the inhibition of biogas production and more specifically the effect of para substituted anilines functional groups (chemical structure) on methane biosynthesis by the digested pig manure methanogens. The objective of this study was also to examine the structure-toxicity relationships of aromatic compounds to acetoclastic methanogens.

Study Design: Anaerobic digestion of pig manure, anaerobic toxicity essay, The effects of functional group nature on inhibition of methane production by acetoclastic methanogens. Correlation of the methanogenic toxicity (IC50) with aromatic compounds hydrophobicity (logPoct).

Place and Duration of Study: Department of Chemistry, University of Kinshasa (DR Congo), between August 2011 and May 2012.

Methodology: The toxicity to acetoclastic methanogenic bacteria was performed with the standard method of serum bottles, digested pig manure was utilized as inoculums, acetate as substrate and the methane gas volume produced was measured by serum bottles liquid displacement systems (Mariotte flask system).

Results: The obtained results indicate that relationships exist between para substituted anilines functional groups nature (chemical structure) and their inhibitory effects on methanogens. The toxicity of para bisubstituted anilines increases in the following order:

                                             SO3 < OH < H < CH3 < Cl < NO2   

From this sequence of increasing toxicity, it can be seen that the methanogenic toxicity varies with the functional group nature which is in the para position of the main function. Indeed, p-Nitroaniline and benzene with 45.76 and 208.78 mg/l as IC50 values respectively were the most toxic compounds, while p-Aminophenol and p-Aminosulfonic acid (Sulfanilic acid) with 1800.39 and 2777.82 mg/l IC50 values were the less toxic.

A very significant negative linear correlation between the toxicity of para substituted anilines compounds and their hydrophobicity was found.

Conclusion: The results of this study indicate that relationships exist between para substituted anilines functional groups nature and their inhibitory effects in methane biosynthesis by the methanogens.

Open Access Original Research Article

Glutathione S-Transferase Enzyme (GSTT1 and GSTM1) Gene Polymorphisms and Oxidative Stress in Egyptian Patients with End Stage Renal Disease

Afaf EL-Said, A. F. AbdeL-Aziz, Basma AL-Sayed Ebead

International Journal of Biochemistry Research & Review, Page 315-327
DOI: 10.9734/IJBCRR/2013/4464

Glutathione S-transferase enzyme (GSTT1 and GSTM1) gene polymorphisms have been associated with the genetic susceptibility to end stage renal disease (ESRD) in different populations. We investigated the association between GSTT1 and GSTM1 genes, and ESRD in Egyptian population. The samples of 133 ESRD and 91 control subjects were collected, and their clinical characteristics were assayed. Glutathione S-transferase enzyme (GSTT1 and GSTM1) gene polymorphisms were detected by polymerase chain reaction (PCR). Serum level of malondialdehyde (MDA), the oxidative stress and lipid peroxidation biomarker, and plasma glutathione S-transferase enzyme (GST), the antioxidant enzyme, were estimated in the ESRD patients as well as in the control subjects. We demonstrated the association of MDA and GST enzyme levels with GSTT1 and GSTM1 genotypes. We investigated the association between MDA and lipid parameters in the ESRD patients. Increased of the GSTM1 deletion genotype, (GSTT1/0) and both deletion genotypes (0/0) in the ESRD patients when compared with the control subjects (P < 0.0001, OR = 3.786, 95% CI = 2.151-6.664), (P = 0.001, OR = 3.172, 95% CI = 1.595-6.308) and (P = 0.045, OR = 1.945, 95% CI = 1.009-3.749), respectively. Highly significant increase of MDA level in the ESRD patients as compared with the control subjects (P < 0.0001). Highly significant decrease of GST enzyme level in the ESRD patients as compared with the control subjects (P < 0.0001).The level of MDA is significantly increased in all GST genotypes in the ESRD patients as compared with the control group. Also, there were significant association between The genetic risk factors of GSTT1 and GSTM1 genes in the ESRD, and high level of MDA in the ESRD patients, while the level of GST enzyme is significantly decreased in GST genotypes except the both deletion (0/0) genotypes, in which the level of GST enzyme is not significantly decreased in the ESRD patients as compared with the control subjects. There are significant association between the genetic risk factors of GSTT1 and GSTM1 genes in the ESRD, (GSTM1 null and GSTT1/0 genotypes), and low level of GST enzyme in the ESRD patients. There were significant positive correlation between MDA and total cholesterol, triglyceride, and LDL-cholesterol, and significant negative correlation between MDA and HDL-cholesterol in the ESRD patients as compared with the control subjects. In conclusion, the GSTM1 (null) genotype, (GSTT1/0) and (0/0) genotypes are independent risk factors for ESRD. The oxidative stress and lipid abnormalities are associated with ESRD in the studied Egyptian population.

Open Access Original Research Article

Role of Human Beta Defensin-2 and Oxidative Stress Parameters in Ovarian Cancer Patients

A. F. Abdel-Aziz, A. A. El-Refaeey, A. Negm, Hanan M. A. El-Taweel

International Journal of Biochemistry Research & Review, Page 364-379
DOI: 10.9734/IJBCRR/2013/5105

Aims: To determine the role of human beta defensin-2 and oxidative stress by measuring serum human beta defensin-2 and antioxidant parameters in ovarian cancer patients.

Study Design: Serum human beta defensin-2 (HBD-2), and the levels of antioxidants such as serum superoxide dismutase (SOD) and catalase (CAT), as well as blood reduced glutathione (GSH) and serum total antioxidant capacity (TAC) and serum malondialdhyde (MDA) were estimated in the circulation of 29 ovarian cancer patients and 15 of age-matched normal subjects as control.

Place and Duration of Study: Department of Gynecology, Mansoura University Hospital, between May 2011 and November 2012.

Methodology: We included 29 patients (all women; age range 20-76 years) with ovarian cancer and the control group comprised 15 age-matched women free from diseases (age range 22-65 years)and was recruited from the gynecology outpatient clinic, Mansoura University. Exclusion criteria were lack of informed consent, patients with associated gynecologic malignancies like cervical, uterine, breast cancers, preexisting immunological as rheumatoid arthritis, psoriasis, Crohn's disease, smoking, and other associated malignancies as colonic, lung carcinoma. Also, patients with any concomitant illness such as obvious systemic infection, diabetes mellitus, hypertension, and renal diseases prior chemo- or radiotherapy, or using corticosteroid therapy were excluded.

Results: A highly significant lowered levels of human beta defensin-2 (.85 +/- .26 ng/ml, P=0.001), catalase activities (504.98 +/- 107.65 U/L, P=0.001), reduced glutathione levels (7.24 +/- 5.36 mg/dl, P=0.001) and total antioxidant capacity levels (1.53 +/- .24 mmol/L, P=0.001) compared with controls (2.74 +/- .92 ng/ml, 717.57 +/- 67.32 U/L, 14.79 +/- 4.29 mg/dl and 2.10 +/- .27 mmol/L respectively). On the other hand, highly significant increased in the concentration of malondialdhyde (9.36 +/- 3.30 mmol/ml, P=0.001) and significantly increased of superoxide dismutase % inhibition (56.70 +/- 9.23 %inhibition, P=0.044) were observed in ovarian cancer patients as compared with controls (6.00 +/- 2.06 mmol/ml and 49.69 +/- 16.83 %inhibition respectively).

Conclusion: The results would suggest that lower human beta defensin-2 as well as oxidative stress may be putative factors in the pathogenesis of ovarian cancer.

Open Access Original Research Article

Peptides Corresponding to Intracellular Regions of GPCR as a New Generation of Selective Drugs

Alexander O. Shpakov

International Journal of Biochemistry Research & Review, Page 380-400
DOI: 10.9734/IJBCRR/2013/5593

In the G protein-coupled receptors (GPCRs) relatively short regions of their intracellular loops and cytoplasmic C-terminal domain are responsible for specific interaction with G-proteins. GPCR-derived peptides corresponding to these regions are able to influence the activity of signal pathways involving the cognate receptors. Modified by hydrophobic radicals, these peptides turn into their cell-penetrating analogs, pepducins, possessing the activity both in vitro and in vivo. This review is devoted to the analysis of the available data and the prospects for GPCR-peptides and their lipophilic derivatives to be used in experimental and clinical medicine in the treatment of vascular diseases, cancers, inflammation, and endocrine dysfunctions.

Open Access Original Research Article

The Effect of COD/N Ratios and pH Control to Biogas Production from Vinasse

S. Sumardiono, I. Syaichurrozi, Budiyono ., S. B. Sasongko

International Journal of Biochemistry Research & Review, Page 401-413
DOI: 10.9734/IJBCRR/2013/3797

Aims: Vinasse that is bottom product of distillation unit from alcohol industry contains Chemical Organic Compound (COD) in high concentration. The purpose of this study was to investigate the effect of COD/N ratios of substrate and pH control to biogas production from vinasse.

Study Design: This study used anaerobic digestion-laboratory scale at room temperature in batch system. Urea was added as nitrogen source to adjust COD/N ratios of 400/7, 500/7, 600/7, and 700/7. Initial pH for all variables was adjusted 7.0 by using NaOH solution.

Place and Duration of Study: Waste-treatment Laboratory, Department of Chemical Engineering, University of Diponegoro, Indonesia, between August 2012 and January 2013.

Methodology: Vinasse used was obtained from the alcohol industry that produced alcohol from molasses. Polyethylene bottles which had volume 5 liters were used as digester. Vinasse of 1 liter was put into digester. Urea was added to make variation of COD/N ratio. Initial pH of all variables was adjusted 7.0 by using 10 N NaOH solution. Rumen fluid (10%v/v vinasse) was added as methanogenic bacteria inoculum. Biogas formed was measured by using water displacement method every once in two days to know biogas production daily. pH of substrates in the digesters was measured by using pH meter every once in two days to know pH profile daily. COD of substrates was measured by using COD Analyzer Hanna Reactor with specification High Range of Reagent. Solution of NaOH 2 N was used to maintain pH of substrate in range neutral condition (7.0±0.2) during the fermentation process.

Results: Biogas formed at COD/N ratio of control variable (1436/7); 400/7; 500/7; 600/7; 700/7 were 3.673; 4.909; 6.079; 6.096; 5.631 mL/g COD respectively. pH profiles for all variables were decreasing from beginning until ending of fermentation. With controlled pH, pH of substrates was maintained at neutral condition, so methanogenic bacteria could grow well in the digesters. Consequently biogas formed at controlled pH was larger than that at uncontrolled pH. The values of COD removal for COD/N ratio of control variable; 400/7; 500/7; 600/7; 700/7 were 1.27±0.43; 1.59±0.43; 2.85±0.39; 3.21±0.49; 2.22±0.39 % respectively at uncontrolled pH, whereas at controlled pH the values of these were 11.98±0.56; 12.82±0.56; 12.03±0.94; 13.05±0.35; 12.61±0.56 % respectively. 

Conclusions: COD/N ratios of 400/7, 500/7, 600/7, 700/7 produced more total biogas than control variable. Variable of COD/N of 600/7 generated the most total biogas which was 6.096 mL/g COD. Biogas production at pH control was greater than that at pH non-control. At non-controlled pH, COD/N ratio of 600/7 had COD removal 3.21±0.49%. Whereas at controlled pH, COD/N ratio of 600/7 had COD removal 13.05±0.35%.  

Open Access Original Research Article

Effect of Regular Physical Exercise (Among Circus Athlets) on Lipid Profile, Lipid Peroxidation and Enzymatic Antioxidants

Shashikant Nikam, Padmaja Nikam, Arun Joshi, R. G. Viveki, A. B. Halappanavar, Bhagyashri Hungund

International Journal of Biochemistry Research & Review, Page 414-420
DOI: 10.9734/IJBCRR/2013/4711

Aims: The study was planned to see the effect of regular physical exercise on levels of lipid profile, activity of lecithin cholesterol acyl transferase, lipid peroxidation and antioxidant enzymes in those involved in regular physical exercise (among athlete in circus) and those involved in sedentary lifestyle.

Study Design:  Cross sectional study.

Place and Duration of Study: The study was carried out during September 2012 to December 2012 in the department of biochemistry, BIMS, Belgaum.

Methodology: A total of 70 participants were studied, aged 25-55 years. Group I consists of 35 participants working as circus athletes, involved in regular physical exercise. Group II consisted of 35 age and sex matched healthy controls, with sedentary life style. Total cholesterol and HDL cholesterol were measured by CHOD-PAP method. Triglyceride was measured by GPO-PAP method. LDL and VLDL were calculated by formula. MDA was determined as the measure of thiobarbituric acid reactive substances (TBARS). SOD Catalase and GPX activity was determined by the method of Mishra and Fridovich, Beer and Seazer and Paglia and Valentine respectively.  

Results: The levels of lipid peroxide, TC, TC / HDL and LDL / HDL ratio were significantly lowered in Group I on comparison with Group II. The levels of HDL, activity of SOD, GPX and catalase were significantly higher in Group I on comparison with Group II. Individuals engaged in regular physical activity had lowered levels of atherogenic lipid components and subjects with sedentary lifestyles had higher atherogenic lipid components (lipid profile and lipid peroxide).

Conclusion: The study indicates that engaging in regular physical exercise protects cardiovascular diseases by increasing the HDL Cholesterol levels, activity of LCAT enzyme and activity of antioxidant enzymes. Thus regular physical exercise is an important in the protection of atherosclerosis and management cardiovascular disease.

Open Access Original Research Article

Structural Mapping of Inhibitors Binding Sites on P-glycoprotein: Mechanism of Inhibition of P-Glycoprotein by Herbal Isoflavones

Neha Arora, Shakti Sahi, Nagendra Singh

International Journal of Biochemistry Research & Review, Page 421-435
DOI: 10.9734/IJBCRR/2013/3268

Aims: Understanding the pattern of inhibitors binding to p-glycoprotein (Pgp).

Study Design: Pgp is an ATP dependent transporter protein, responsible for multi-drug resistance in metastatic tumors. It removes toxins by exporting a variety of structurally unrelated compounds outside the cells, which make Pgp a promising target for designing anti cancer supplementary therapeutic molecules. Isoflavones are present in soyabean and other herbal extracts. The idea was to explore inhibitor binding sites on Pgp to find hotspots which eventually may prove useful in designing compounds with higher specificity and affinity.

Place and Duration of Study: School of Biotechnology, Gautam Buddha University, Greater Noida, between February 2012 and December 2012.

Methodology: The biochemical nature of binding of isoflavones to Pgp has been extensively studied, but the atomic details of their interactions were not understood. Therefore, we have used in silico methods to study binding of eleven isoflavones to Pgp. The docking studies were performed using grid-based ligand docking with energetic (GLIDE).

Results: Isoflavones binds at two slightly distinct sites perpendicular to each other, present in the large hydrophobic cavity of Pgp. Three isoflavones bind to site 1, whereas eight isoflavones bind to site 2 by forming van der Waals and H-bonded interactions. Both the sites are highly hydrophobic in nature and are contributed mainly by side chain of non polar residues present on twelve transmembrane a-helices. Site 1 has minimum dimension of 7.5Å and maximum as 22Å whereas, site 2 is wider and deeper than site1. One sidewall of the site 2 is formed by polar amino acid residues of helix H12, which makes several hydrogen bonds with ligands.

Conclusion: Structure analysis revealed that addition of polar group to hydrophobic ligand may enhance its binding affinity for Pgp, which may be used for designing potent inhibitors to find lead compounds for drug design.

Open Access Review Article

Evolution of Rodent Pheromones: A Review of the ABPs with Comparison to the ESPs and the MUPs

Robert C. Karn

International Journal of Biochemistry Research & Review, Page 328-363
DOI: 10.9734/IJBCRR/2013/5763

Three proteinaceous pheromone families, the androgen-binding proteins (ABPs), the exocrine-gland secreting peptides (ESPs) and the major urinary proteins (MUPs) are encoded by large gene families in the Mus musculus and Rattus norvegicus genomes. The purpose of this article is to review what is known about the evolutionary histories of the the Abp gene family expansions in rodents and, where appropriate, to compare them to what is known of the expansions of the Mup and Esp gene families. The issues important to these histories are the extent of the gene family expansions, the timing of their expansions and the roles played by selection, gene conversion and non-allelic homologous recombination (NAHR).  I also compare and contrast the evolutionary histories of all three mouse gene families in light of the proposed functions of their pheromones in mouse communication.