Open Access Original Research Article

Polyprenols of Grape Vitis vinifera L. Leaves

U. T. Zakirova, N. M. Mamatkulova, Kh. U. Khodjaniyazov, N. K. Khidyrova, Kh. M. Shakhidoyatov

International Journal of Biochemistry Research & Review, Page 97-106
DOI: 10.9734/IJBCRR/2013/2935

Aims: The study is aimed to isolate of polyprenols from grape Vitis vinifera L. leaves by different methods and to determine of polyprenol content of plant leaves growing in various regions of Uzbekistan.

Study Design: Isolation of polyprenols.

Place and Duration of Study: Department of Organic Chemistry, Institute of the Chemistry of Plant Substances (ICPS). The study was carried out between January, 2011 and December, 2012.

Methodology: We have studied isolation of polyprenols of grape leaves by using of simple, microwave and ultra sound extraction. Spectral, HPLC and HPTLC methods have used.

Results: It was determined that leaves polyprenols are in type polyprenyl homologues with 10-13 isoprene units where undecaprenol and dodecaprenol were dominant. Extraction carried out of grape leaves in various conditions (usual, microwave, ultrasound). Ethanol was high effective for microwave and ultrasound extraction. It was observed forming of polyprenol and high aliphatic alcohols esters (probably proceeding particularly esterification of polyprenols and aliphatic alcohols by organic acids which contain in extracts).

Conclusion: It was determined that extraction of grape leaves by alcohol in microwave and ultra sound stirring are high effective methods for isolation of polyprenols.

Open Access Original Research Article

Preliminary Studies on the Expression and Purification of Functionally Active Recombinant Plasmepsin 9 from Plasmodium falciparum

Folasade M. Olajuyigbe

International Journal of Biochemistry Research & Review, Page 107-118
DOI: 10.9734/IJBCRR/2013/3023

Aim: Plasmepsins, a group of homologous aspartic proteinases are attractive drug targets against malaria. Plasmepsin 9 (PM9) expressed in the blood stage of malaria life cycle with unknown function has been strongly considered as a potential target. However, recombinant expression of active PM9 for biochemical and structure-activity analysis of the enzyme has been very challenging. This paper presents preliminary report on the expression and purification of active recombinant plasmepsin 9 from Plasmodium falciparum.

Place and Duration of Study: Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, Florida between November 2011 and October, 2012.

Methodology: The plasmid of truncated proenzyme (proPfPM9) from Plasmodium falciparum was expressed in Escherichia coli Rosetta 2(DE3)pLysS competent cells. The protein was purified from inclusion bodies using combination of cation exchange and gel filtration chromatography. The expression and purification fractions were subjected to SDS-PAGE. The zymogen was activated to produce mature form of active recombinant PfPM9. Catalytic activity test of PfPM9 was determined using a chromogenic substrate, Lys-Pro-Ile-Glu-Phe-Nph*Arg-Leu (RS6). Substrate hydrolysis was examined from substrate cleavage scan. 

Results: SDS-PAGE confirmed the expression and purification of truncated proenzyme (proPfPM9) with molecular weight of approximately 54 kD. Kinetic assays showed that the purified PfPM9 was active. The hydrolytic activity of PfPM9 on RS6 was confirmed by a shift in absorbance peak from 280 nm to 272 nm at the end of assay reactions. Inhibition studies on the mature PfPM9 showed that the activity of the enzyme was inhibited by pepstatin A.

Conclusion: Protocols for successful expression and purification of active recombinant plasmepsin 9 in E. coli have been established in this preliminary report. These will aid production of sufficient yield of PfPM9 for detailed kinetic and structural characterization of the enzyme and development of future inhibitors against malaria.

Open Access Original Research Article

Toxic Effects of Sub-Chronic Administration of Chloroform Extract of Artemisia maciverae Linn on the Kidney of Swiss Albino Rats

Atawodi Sunday Ene-ojo, Ene Aloy Chinedu, Fatihu Muhammad Yakasai

International Journal of Biochemistry Research & Review, Page 119-128
DOI: 10.9734/IJBCRR/2013/2062

The nephrotoxic potential of sub-chronic doses of chloroform extract of Artemisia maciverae Linn was studied in male Swiss albino rats. The groups were respectively administered chloroform extract of Artemisia maciverae at 0, 50, 100 and 200 mg/kg b.wt for 60 days and then monitored till day 90 before sacrifice. Sera samples were analyzed for urea and creatinine. The kidneys were subjected to histological examination after staining with hematoxyline-eosin. At the onset of treatment, the extract caused statically significant (p<0.05) elevation in serum urea and creatinine. The mean (+SD) levels of serum urea at the onset of treatment with 0, 50, 100 and 200 mg/kg of the extract were 29.6+ 1.10, 54.1+4.40, 81.6+8.50 and 132.1+6.10mg/dL respectively, while that of serum creatinine were 0.5+0.10, 0.8+0.10, 1.2+0.10 and 1.3+0.30mg/dL respectively. The elevations in serum urea levels returned to normal after the onset of treatment, but that of creatinine persisted. Thirty days after withdrawal of treatment, the levels of serum urea in the 0, 50 and 100 mg/kg treatment groups were found to be 30.1+2.00, 32.1+ 2.00 and 33.3+1.30mg/dL respectively, while that of serum creatinine were found to be
0.6+0.10, 1.2+0.10 and 1.2+0.10mg/dL respectively. Microscopically, tubular epithelial necrosis was observed in the treated animals in the early stages, but the renal injuries disappeared at the later stage. These results suggest that chloroform extract of Artemisia maciverae may be nephrotoxic at high doses.  

Open Access Original Research Article

Comparative Hypolipidaemic Effect of Aqueous Extract of Psidium guajava Leaves and Ascorbic Acid in Male Rats

F. E. Uboh, G. O. Igile, I. E. Okon, P. E. Ebong

International Journal of Biochemistry Research & Review, Page 129-136
DOI: 10.9734/IJBCRR/2013/2098

Aim: To assess the hypolipidaemic properties of aqueous extract of Psidium guajava leaves (AEPGL) and ascorbic acid (vitC) in male albino Wistar rats.

Study Design: Three groups of rats were respectively administered distilled water, 200mg/kg of AEPGL and vitC orally, for thirty days, to determine the hypolipidaemic properties of AEPGL and vitC. The hypolipidaemic properties were determined from the serum lipid profile of the rats.

Place and Duration of Study: This work was carried out in Biochemistry Department, University of Calabar, Calabar, Nigeria between January and February, 2012.

 Methodology: Eighteen albino Wistar rats, divided in three groups of six rats each, were used in this study. The rats in group I (control) were administered, by oral gavage, with 2ml of distilled water. While the rats in groups II and III received 200/mg/kg body weight oral daily doses of aqueous extract of AEPGL and vitC, respectively, using orogastric tubes for 30 days. The hypolipidaemic properties of AEPGL and vitC were determined from their effect on serum lipid profile of the treated rats, using serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), very low-density lipoprotein cholesterol (VLDL-C), high density lipoprotein cholesterol (HDL-C), TG/HDL-C and artherogenic index of plasma (AIP). Standard reagent kits were used to determine serum TC, TG and HDL-C, while LDL-C, VLDL-C, TG/HDL-C and AIP were calculated.

Result: The results showed that serum HDL-C level obtained for rats treated with AEPGL (7.10 ± 0.11mg/dl) and vitC (6.93 ± 0.11mg/dl) were significantly (p<0.05) increased, compared respectively to that of the control rats (3.73 ± 0.18mg/dl). While the levels of TC, TG, LDL-C, VLDL-C, TG/HDL-C and AIP obtained rats treated with AEPGL (4.63 ± 0.21, 3.45 ± 0.14, -3.15 ± 0.28, 0.68 ± 0.02mg/dl, 0.48 ± 0.02 and -0.32 ± 0.02, respectively ) and vitC (4.68 ± 0.18, 3.65 ± 0.15, -2.98 ± 0.24, 0.73 ± 0.03 mg/dl, 0.53 ± 0.03 and -0.28 ± 0.03, respectively) were significantly (p<0.05) decreased, compared respectively to the control rats (7.22 ± 0.06, 6.87 ± 0.10, 2.12 ± 0.21, 1.37 ± 0.02mg/dl, 1.88 ± 0.08 and 0.27 ± 0.02, respectively). However, the percentage increase in serum HDL-C, and decrease in serum TC, TG, (LDL-C), VLDL-C, TG/HDL-C and AIP associated with administration of AEPGL was observed to be insignificantly (p>0.05) higher than that recorded for vitamin C. Conclusion: In conclusion, the results of this study indicate that AEPGL and vitC possess hypolipidaemic, hence antiatherogenic, properties in male rats, thereby supporting the local use of P. guajava leaves in the management of hypertensive conditions.

Open Access Original Research Article

Phytochemical Studies, In-vitro Antibacterial Activities and Antioxidant Properties of the Methanolic and Ethyl Acetate Extracts of the Leaves of Anogeissus leiocarpus

Barku Y. A. Victor, Abban Grace

International Journal of Biochemistry Research & Review, Page 137-145
DOI: 10.9734/IJBCRR/2013/3349

Aims: To conduct a study on the antibacterial activity, phytochemical constituents and antioxidant properties that identify the medicinal potency of medicinal plants on the methanolic and ethyl acetate extracts of the leaves of Anogeissus leiocarpus. Antibacterial activity was determined by well diffusion method against four bacterial strains commonly associated with wound infections.

Study Design: Standard Phytochemical quantitative and qualitative test, DPPH radical scavenging activity and well diffusion method.

Place and Duration of Study: Department of Chemistry (Natural Products Laboratory), University of Cape Coast and Department of Microbiology, Korle-Bu Teaching Hospital, University of Ghana, between March 2012 and January 2013.

Methodology: Standard chemical procedures were used to identify phytoconstituents present in the plant extracts. DPPH radical scavenging assay and well diffusion method were used to determine antioxidant and antibacterial activity respectively. UV-spectrophotometry was employed to further quantify the phenolic and flavonoid content.

Results: The extracts showed pronounced activity against all the bacterial strains tested.

Ethyl acetate exhibited the maximum zone of inhibition against Klebsiella pneumonia while the Methanol extract exhibited the maximum zone of inhibition against Citrobacter sp. All the extracts showed higher sensitivity against Gram negative bacterial strains than the Gram positive bacterial strain used in this study. Phytochemical study was performed by using standard methods. Quantitative estimation of bioactive phytoconstituents showed that the plant contains alkaloids (152.0 ± 0.1 mg/g), phenolics (1294.81 ± 3.0 mg/g), flavonoids (330.7 ± 3.0 mg/g) in the methanol extract and alkaloids (80.20 ± 0.0 mg/g), phenolics (616.5 ± 4.4 mg/g), flavonoids (202.5 ± 4.0 mg/g) in the ethyl acetate extract respectively. In similar fashion, the results from the scavenging activity against 1,1-diphenyl-2-picryl hydrazyl radical (DPPH) and the Ferric reducing antioxidant power (FRAP) assay for both extracts were high and concentration dependent.

Conclusion: The study clearly indicates that A. leiocarpus possesses significant antioxidant activity and antibacterial activity against the tested wound infectious bacterial strains. Hence constituents of the plant could be studied in the aim to be used in chemotherapy.

Open Access Review Article

Chronological Primacy of Oxidative-Induced Neuronal Damage in the Pathogenesis of Alzheimer's Disease

Selim F. Estefan, Aziza B. Shalby, Hanaa H. Ahmed

International Journal of Biochemistry Research & Review, Page 82-96
DOI: 10.9734/IJBCRR/2013/2593

Alzheimer’s disease (AD) is associated with hallmark pathologies including extracellular Aβ protein deposition in extracellular senile plaques and vessels, and intraneuronal tau deposition as neurofibrillary tangles.

The current study comprises the oxidative modifications associated with the pathological lesions of neuronal damage characterized AD. The repeated exposure to aluminum and heavy metals, mutations in a number of chromosomes and genes, diabetes, cardiovascular diseases, obesity and brain injury, are the major causes for these modifications.

There is clearly a need for the identification and development of panels of biomarkers for accurate diagnosis and early detection of sporadic AD. Thus, a collection of the most globally manifested expeditious diagnostic tools for early detection of AD is outlined in this review. Also, a survey of the vast clinically approved therapeutic modalities for restricting and even treating the symptoms of AD is summarized. These arguments provide useful information in both understanding pathogenesis as well as accessing the novel treatment approaches for AD.