Open Access Original Research Article

Establishment of cDNA Library and EST Analysis from Leaves of Phyllanthus amarus

Sharmila Chattopadhyay, Aparupa Bose Mazumdar Ghosh

International Journal of Biochemistry Research & Review, Page 1-15
DOI: 10.9734/IJBCRR/2014/5262

DNA sequencing of randomly chosen clones from a cDNA library allows thousands of different transcripts to be identified. However, since the likelihood of observing a given transcript is proportional to the expression level of that transcript in the tissue from which the library is derived, often transcripts are represented by several EST sequences. An expressed sequence tags (EST) analysis was undertaken to identify the genes present in the leaves of Phyllanthus amarus, which is a small tropical, glabrous herb with several health benefits. Phyllanthin and hypophyllanthin, major bioactive components, present in highest amounts in the leaves, are of significant therapeutic importance like hepatoprotective, antioxidant, antiviral, hypoglycemic, etc. Taken together, sequencing of cDNA clones generated high-quality ESTs (Accession number: JK492908 to JK492964) with high similarities with genes from Ricinus communis, Onchocerca volvulus, Eucalyptus globules, Gossypium hirsutum, Nicotiana tabacum, Solanum spp. and many more. A BLASTN analysis along with BLASTX analysis of all the unique sequences was performed and was grouped according to the reported activities. Results represented here is the first reference collection of ESTs from this commercially important medicinal herb. This study indicated that the leaf transcriptome contains series of interesting sequences like ALBINO3, ribulose-1, 5 bisphosphate carboxylase/ oxygenase (RUBISCO), chloroplast photosystem II chlorophyll A/B-binding protein, stress-responsive proteins like methionine sulfoxide reductase type, etc.

Open Access Original Research Article

Evaluation of the Concentration of Selected Heavy Metals and the Effects on Soil Enzymatic Activities in an Abandoned Cement Factory Nigercem Nkalagu and Its Environs

Orinya Onyebuchi Frederick, Akubugwo Emmanuel Iroha, Edeogu Chukwu Oswald

International Journal of Biochemistry Research & Review, Page 16-27
DOI: 10.9734/IJBCRR/2014/5019

This study investigated the concentrations of heavy metals, the activities of some selected enzymes and the pH of the soil surface (1-10 cm) and depth (11-20 cm) within the vicinity of an abandoned cement factory Nigercem Nkalagu, and further away 1km from the factory centre. The results indicate that the pH and heavy metals gradually decreased with increase in distance from the cement factory centre with the exception of Fe, Zn and Pb which was the reverse. The concentrations were also higher in the depths. The enzymatic activities increased away from cement factory centre and decreased with depth. There was a significant difference (p ≤0.05) between most of the sampling points and the control (1 km) for all the parameters studied, except Cd and Cr. The enzymatic activities correlated positively with Fe, Zn and Pb and negatively with the other parameters. The inverse correlation between most of these heavy metals, the pH, and the activities of the soil enzymes are indicative that pollution caused by cement production still exhibit a significant effect on the enzymatic activities and this may invariably affect the quality of the soil of this area.

Open Access Original Research Article

Preparation, Characterization, Theoretical Treatment and Adsorption on SurfacesN5AcyclicImmobilizes on Silica Gel for Removal of Some Toxic Metals

Omar Hamad Shehab AL- Obaidi, Mahmoud N. Al-Jibouri

International Journal of Biochemistry Research & Review, Page 28-42
DOI: 10.9734/IJBCRR/2014/5198

A series of transition elements complexes with [2,6-diacetimino-pheylenediamine –[2,2,diyl] pyridine] of the general formula [MLCl]Clx, (where M= Cr(III), Pb(II), Zn(II) and Cd(II), x= 2 for Cr(III)) respectively have been synthesized  and  fully characterized  on the bases of C.H.N.M, elemental analysis, 13C NMR, UV- Visible and FTIR spectra , in addition the structure of complexes was characterized by magnetic moments and molar conductance in DMF solution ,and molar ratio of complexes  were also determined. On the basis ofelectronic spectral studies .All complexes have an octahedral geometry around the metal center in (1:1) (L:M).

A theoretical treatment of the formation of complexes in the gas phase was studied, this was done using the HYPERCHEM-6 program for the Molecular mechanics and Semi-empirical calculations. The batch adsorption method has applied on N5 polydentate ligand to remove the selected toxic metal ions from waste water after immobilization the newly n5-ligand on silica gel by physic-chemical method.

Open Access Original Research Article

Cross Talk between Oxidative Stress, PPAR- γ and RAS System in Diabetic Rat Heart Exposed to Ischemia / Reperfusion Injury

Maha M. Sabry, Nagwa K. Roshdy, Laila A. Rashed, Hany E. ElSebaee, Mary A. Youssef, Amal F. Tawadrous

International Journal of Biochemistry Research & Review, Page 66-79
DOI: 10.9734/IJBCRR/2014/6009

Introduction: The present study aimed at clarifying the cross talk between peroxisome  proliferator-activated receptor-gamma (PPAR- γ), cardiac reactive Oxygen species (ROS) and Renin-Angiotensin System (RAS).

Methods: A total of 90 male albino rats were used. The rats were divided into: Group 1: Control group, Group 2: Type 2 diabetic rats, Group 3: PPARγ agonist protected type 2 diabetic rats. Group 4: Antioxidant protected type2 diabetic rats, Group 5: Metformin treated type 2 diabetic rats. Blood samples were collected for measurement of FBS and fasting insulin. Half the number of each group was sacrificed and the heart excised and perfused, from the rest of the group small piece from the heart was taken for estimation of malondialdehyde (MDA), Angiotensin 2 Receptor (AT2R) and Angiotensin Converting Enzyme 2 (ACE2) gene expression.

Results: Treatment with pioglitazone and Vitamin E significantly lowered blood glucose, insulin levels and Homeostasis Model Assessment Insulin Resistance (HOMA IR). However, values did not return to control values. Pioglitazone and Vitamin E improved myocardial performance and percentage recovery following ischemia reperfusion. The cardioprotective effect was more pronounced in the pioglitazone group. This positively correlated with decreased MDA levels and increased AT2R and ACE2 expression in cardiac tissue. 

Conclusion: Pioglitazone and Vitamin E in type 2 DM significantly offered cardio-protection through improving the diabetic condition and / or decreasing MDA levels.

Open Access Original Research Article

Effects of Aegopodium podagraria Preparations on the Metabolic Disorders Induced in Rats by Excess Fructose Combined WITH Hydrochlorothiazide: The Relationship between Influence on Electrolyte and Carbohydrate Metabolism

Olga Tovchiga

International Journal of Biochemistry Research & Review, Page 80-98
DOI: 10.9734/IJBCRR/2014/7201

Aims: To evaluate the effect of Aegopodium podagraria aerial part extract and tincture on electrolyte, glucose and uric acid metabolism in rats receiving excess fructose (10% drinking solution) combined with hydrochlorothiazide (20 mg/kg).

Study Design: Fourty-two male rats were randomly and evenly distributed to seven groups: Group I: intact control; Group II: fructose supplementation; Group III: fructose supplementation + hydrochlorothiazide; Group IV: fructose supplementation + hydrochlorothiazide + A. podagraria extract, 1 g/kg; Group V: fructose supplementation + hydrochlorothiazide + A. podagraria tincture, 1 ml/kg; Group VI: fructose supplementation +hydrochlorothiazide +A. podagraria tincture, 5 ml/kg; Group VII: fructose supplementation + hydrochlorothiazide + reference drug allopurinol, 10 mg/kg. The duration of the treatment was 10 weeks.

Place and Duration of Study: Central Scientific-Research Laboratory of National  University of  Pharmacy, Kharkiv, Ukraine, between February and 2010 and May 2010.

Methodology: At week 9, glucose tolerance test was performed. At week 10, the excretory renal function was analyzed, creatinine, uric acid, sodium and potassium level in urine and plasma were determined, glomerular filtration rate, sodium and water reabsorption, Na+ /K+ ratio of urine and plasma were calculated.

Results: Hydrochlorothiazide against the background of fructose increased sodium excretion and urine Na+ /K+ ratio (P=.05). The extract and the tincture (5 ml/kg) tended to increase sodium reabsorption while the tincture at the lower dose and allopurinol did not change the renal function. The extract exerted a hypouricemic effect (P=.05). In contrast to the tincture and allopurinol, the extract augmented potassium excretion and reduced its blood content (P=.05). Base on Na+/K+ ratios these changes may be associated with unfavourable hyperaldosteronism. The еу ерtincture (1 ml/kg) decreased blood glucose level (P=.05).

Conclusion: A. podagraria tincture exerts hypoglycemic effect on the model similar to metabolic syndrome. Further study of A. podagraria extract dosage regimens is required.

Open Access Original Research Article

Comparative Effects of Aloe vera Gel and Aqueous Leaf Extract of Viscum album on Bilirubin Excretion in Streptozotocin - Induced Diabetic Rats

Nna Victor Udo, Ofem Ofem Effiong, Oka Victor Otu, Aluko Esther Olusola, Ofutet Emmanuel Oleba

International Journal of Biochemistry Research & Review, Page 99-115
DOI: 10.9734/IJBCRR/2014/7512

Aims: This study was carried out to determine the effect of type 1 Diabetes Mellitus on bilirubin excretion and to compare the effects of separate administration of Aloe vera gel and aqueous leaf extract of Viscum album on serum bilirubin, bile secretory rate and biliary bilirubin concentration.

Methodology: Thirty six male albino Wistar rats weighing 180 - 220 g were used for this study. After 14 days of habituation, the rats were randomly divided into 6 groups of 6 rats each. Type 1 Diabetes Mellitus was induced in the test groups by a single i.p dose (65 mg/kg) of streptozotocin. Group 1 served as control; group 2 - diabetic untreated group (DM); group 3 - diabetic group, treated with 0.4 ml/100g Aloe vera gel orally (DM+Aloe); group 4 - diabetic group, treated with 150 mg/kg Viscum album leaf extract orally (DM+VA); group 5 - control group, treated with 0.4 ml/100g Aloe vera gel orally (C+Aloe); group 6 - control group, treated with 150 mg/kg Viscum album leaf extract orally (C+VA). All animals had unrestricted access to food and water. The regimen lasted for 21 days, after which bile secretion was determined and same was collected together with serum for biliary and serum bilirubin estimation. 

Results: The results showed that serum and biliary total, conjugated and unconjugated bilirubin concentrations  were significantly (p<0.001) higher in the DM group compared to control, DM+Aloe and DM+VA, with DM+Aloe group having significantly lower serum and biliary total and conjugated bilirubin (p<0.001), and serum unconjugated bilirubin (p<0.05) compared to DM+VA group. Serum conjugated bilirubin concentration in C+Aloe and C+VA group was significantly (p<0.05 and p<0.001 respectively) higher compared to control, while serum unconjugated bilirubin concentration was significantly (p<0.001 and p<0.01 respectively) lower compared to control. C+VA group had a significantly (p<0.001) higher biliary total and conjugated bilirubin concentrations compared to C+Aloe group.

Conclusion: On the basis of the results obtained, we therefore conclude that Aloe vera gel and aqueous leaf extract of Viscum album enhances bilirubin excretion in diabetic and normal animals and are both hepatoprotective.

Open Access Review Article

History and Biology of Transglutaminase 2: A Synopsis

Odii Benedict Onyekachi, Peter Coussons

International Journal of Biochemistry Research & Review, Page 43-65
DOI: 10.9734/IJBCRR/2014/6363

Transglutaminase 2 (TG2) belongs to the family of transglutaminase, a large group of intracellular and extracellular enzymes that primarily catalyze the Ca2+-dependent post-translational modification of proteins. Discovery of the first transglutaminase in the early 1920s, has subsequently lead to the identification of nine members of the enzyme family including TG2; the most abundant, most popular and most studied member of the transglutaminase enzyme family. The popularity of TG2 is due to its uniqueness amongst other members of the Transglutaminase (Tgase) family. Its difference from other Tgase family members is due to its specialized structural and biochemical activities; abundant tissue distribution and sub-cellular localization; and multi-functionality and physiology. The growing interests in TG2 and related research has resulted to an attendant mega-research output; and the need to produce a well-structured compilation of data on this popular enzyme has arisen. It is against this background, that we have compiled herein, a synopsis of available literature on TG2 history, structural and biochemical activities, tissue distribution, and physiology. This was done with the view to providing a compendium of background information that could be handy to researchers and new interest in the field of TG2 research.