Open Access Short Research Article

Premna integrifolia L. on Enzymatic Biomarkers in Atherosclerosis

S. Chitra, R. Arivukkodi, S. Gaidhani, R. Ilavarasan, K. S. Dhiman Vd

International Journal of Biochemistry Research & Review, Page 1-9
DOI: 10.9734/IJBCRR/2017/34232

Premna integrifolia L. belongs to family Verbenaceae and it is believed to prevent cardiovascular disease as per Ayurveda. In order to evaluate the cardiac biomarkers and treatment efficacy of hydroalcoholic extract (HAE) of root bark in atherosclerosis. Sixty Wistar rats were divided into six groups and ten animals in each group. Rats fed with high fat diet and various concentrations of HAE treatment were studied and standard drug, atorvastatin was compared with HAE. This extract showed a significant action against atherosclerosis in a dose dependent manner which was studied in terms of lactate dehydrogenase activity, HMG-CoA/Mevalonate ratio, collagen content in aorta, levels of transaminases, calcium and other marker enzymes indirectly involved in the cardio protection. Alteration in collagen deposition in aorta of histopathological evaluation was noticed in treated animals. Anti-atherosclerotic activity of HAE of P. integrifolia was due to its modulatory activity on cholesterol metabolism and our results contribute towards validation of the traditional use of P. integrifolia in high fat diet induced atherosclerosis.

Open Access Short communication

Identification, Characterization and Purification of MSC_0265, a Potential Immunogenic Antigen Homologue of Mycoplasma mycoides subsp. mycoides in Mycoplasma capricolum subsp. capripneumoniae

M. Orwe, J. Kinyua, J. Ngaira, H. Wesonga, J. Naessens

International Journal of Biochemistry Research & Review, Page 1-12
DOI: 10.9734/IJBCRR/2017/34125

In silico identification and characterization of vaccine antigens has opened up new frontiers in the field of reverse vaccinology to mitigate the effects of livestock diseases by development of new subunit vaccines. This study aims to characterize, express and purify MSC_0265 for eventual use in immunoassays and inoculation in goats. Mycoplasma mycoides subs. mycoides (Mmm) and Mycoplasma capricolum capripneumoniae (Mccp) are similar pathogens on the genomic level and are the causative agents of Contagious Bovine Pleuropneumonia (CBPP) in cattle and Contagious Caprine Pleuropneumonia (CCPP) in goats respectively. In this study, BLAST was used to identify the homology of MSC_0265 in Mycoplasma capricolum capripneumoniae genome and the protein it is similar to. Characterization of MSC_0265 was also done using I-TASSER to predict secondary structure, solvent accessibility, normalised B-factor, 3D models and function. With cut off points of 0.0 for E-value, 100% for Query coverage and 90% for Identity, MSC_0265 a pyruvate dehydrogenase enzyme gave a high homology score on tBLASTn and BLASTp. It had earlier been cloned in pGS21a vector before proceeding with expression and purification of the His-tagged protein by Ni-NTA affinity chromatography. This study identified the homologue of MSC_0265 as protein WP_029333261.1 in the Mycoplasma capricolum capripneumoniae genome (Accession NZ_LN515398.1) using tBLASTn and BLASTp. Additionally, MSC_0265 was characterized and its optimal expression profile and estimated molecular weight verified.

Open Access Original Research Article

Standardization of Protocol for Best Blending Ratio of Papaya cv. Red Lady and Guava cv. Allahabad Safeda Fruit Pulp for Preparation of Fruit Bar

Appani Laxman Kumar, C. Madhumathi

International Journal of Biochemistry Research & Review, Page 1-10
DOI: 10.9734/IJBCRR/2017/34077

The papaya and guava pulps were blended in the ratios of 100:0, 80:20, 60:40, 50:50 and 40: 60 in preparation of fruit bar and physicochemical analysis and sensory attributes were evaluated at zero (initial), 30 and 60 days of storage. Among the different blending ratios, fruit bar prepared with 60% papaya pulp +  40% guava pulp (T3) was selected as best ratio (combination) as the treatment had maximum score for sensory attributes i.e., overall acceptability (8.75). The microbial load recorded in the same treatment was also minimum (yeast and mould) and within safe levels for consumption even after 60 days of storage. In general there was an increase in total soluble solids, reducing sugars and pH, whereas moisture content, titrable acidity, total sugars, ascorbic acid, total carotenoids, protein content and all sensory attributes were decreased with the advancement of storage period.

Open Access Original Research Article

Quantitation of TLR-2 mRNA Expression in Bovine Mastitis Caused by E. coli

R. Lakshmi, K. K. Jayavardhanan

International Journal of Biochemistry Research & Review, Page 1-5
DOI: 10.9734/IJBCRR/2017/33367

Aims: This study aim was to assay the expression of an important TLR2 gene in naturally infected cows with sub-clinical and clinical mastitis caused by E. coli.

Methodology: cDNA from E. coli infected samples were obtained from hind quarters of three animals with sub-clinical mastitis, three animals with clinical mastitis along with a healthy animal, were used for TLR2 expression analysis using commercial kit and oligo (dT) primers. Quantitative reverse transcription PCR (RT-qPCR) were standardized for the TLR2 and β-actin gene, genes expression level was analyzed by Illumina Eco® Q- RT PCR system.

Results: The mRNA expression of TLR2 gene in animals sub-clinical mastitis was significant (P ≤ 0.01) higher (3.63 fold) than clinical case (1.89 fold) when compared to healthy bovine.

Conclusion: The expression of TLR2 gene during early stage of infection was at high level, therefore most of the sub-clinical mastitis subsided by itself without precipitating into clinical mastitis. It is suggested that TLR2 may consider as candidate gene for mastitis screening in cattle population.

Open Access Original Research Article

Toxicity Assessment of Sub Lethal Doses of Chlorpyrifos on the Kidney and Liver Organs of Male Wistar Rats

O. O. Fayinminnu, S. O. Tijani, O. O. Fadina

International Journal of Biochemistry Research & Review, Page 1-15
DOI: 10.9734/IJBCRR/2017/31349

Chlorpyrifos, an organophosphate pesticide is an important neurotoxic and tissue damage agent. It is one of the most heavily used pesticides in domestic and agricultural applications globally. Repeated doses of chlorpyrifos have been able to cause significant disturbances on the biochemical and physiological functions of the blood, and histological abnormalities in livers and kidneys exposed to this insecticide. The toxicities of sub lethal oral administration of chlorpyrifos daily for 28 days were assessed using a completely randomized design. Twenty five albino Wistar rats weighing between 150-200 g divided into five groups containing five rats each were housed in the Central Animal house of College of Medicine, University of Ibadan. Chlorpyrifos at 0 (control), 18.9, 25.9, 32.2 and 39.2 mg/kg were orally administered to male rats, respectively for four weeks, between the months of May and June, 2014. At the end of the experimental period, the toxicities of chlorpyrifos were assessed in rats using haematology, serum liver enzymes and histopathological assays. Results revealed significant reduction in body weights compared to control. The packed cell volume (PCV), hemoglobin (Hb) and lymphocytes (Lymp) also showed significant reduction at 39.2, 25.9 and 32.2 mg/kg (39.33, 45.40 and 44.80%), respectively compared to control (49.67%). Insecticide administration to rats resulted in significant elevation of serum transaminases (AST and ALT) and alkaline phosphatase (ALP) from 32.2, 39.2 and 25.9 mg/kg dosages, (ranged from 294.94-542.00 u/L for AST and from 96.25-130.77 u/L for ALT), respectively. Also, experimental treated groups exhibited marked of total protein and altered albumin and globulin contents compared to control. Studies revealed dose dependent increase of histopathological alterations. The livers showed moderate vacuolar change of hepatocytes, having a finely reticulated cytoplasmic and congestion of central veins. The kidneys showed  mild focal sloughing off of tubular epithelium of renal cortex, fluid in tubular lumen, tubules appearing dilated and cystic (nephrosis) and proteinaeceous fluid in Bowman`s capsule and compressing the glomerulus tufts (Esinophilic). Different concentrations of chlorpyrifos including the lowest tested dose produced marked alterations in the exposed animals in this study and thereby affecting the overall performance in terms of health and wellbeing. Thus, this could cause similar health and environmental risks to humans even at the lowest dose.