MULTIPLE TREATMENT OF EREMURUS HIMALAICUS EXTRACTS AMELIORATES CARBON TETRACHLORIDE INDUCED LIVER INJURY IN RATS

Objective: Eremurus himalaicus Baker, an edible herb of North Western Himalayas, has not been scientifically assessed for hepatoprotective potential. The ethyl acetate extract (EHE), methanolic extract (EHM) and aqueous extract (EHA) of Eremurus himalaicus were therefore evaluated for potential hepatoprotective activity in Wistar strain albino rats.Methods: Carbon tetrachloride (1.5 ml/kg) was employed as hepatotoxin and was given on Day 1 of the experiment. The extracts at a dose of 300 mg/kg bw (EHE, EHM and EHA) and the standard at a dose of 10 mg/kg bw (Liv 52) were given for following 7 d and the biochemical parameters (SGOT, SGPT, ALP, TP, bilirubin and UA) were estimated in order to assess the liver function. Moreover, the liver tissue samples were examined for histopathological changes.Results: The results for serum biochemical analysis in rats showed a rise in SGOT, SGPT, ALP and bilirubin levels and a decrease in TP and UA levels upon giving hepatotoxin. The administration of the extracts and standard drug, for a period of 7 d, showed a significant decrease in SGOT, SGPT, ALP and bilirubin levels and an increase in TP and UA levels for EHM when compared to the toxic group. These results correlated well with the histopathological findings of liver for normal, toxic and extract treated groups. The EHM treatment decreased the extent of fat deposition and necrosis caused by CCl4. The results were almost similar to the standard drug Liv 52.Conclusion: Collectively; the results indicate that EHM exhibits significant hepatoprotective activity against CCl4 induced hepatotoxicity

Prevalence of iron deficiency Anaemia among adolescent girls and impact of Health and Nutrition Education Programme in changing their dietary behaviour

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Antimicrobial potential of some wild Macromycetes collected from Kashmir Himalayas

Alarming increase in microbial resistance to existing synthetic commercial antibiotics forced scientists to search for new antimicrobials from various alternative sources. The present study carried out during the year 2014-2015, presents the antimicrobial potential of some mushroom extracts against some commonly found pathogenic bacterial and fungal microbes. During the study four mushroom species, viz. Lentinus tigrinus (Bull.) Fr., Fomitopsis pinicola (Sw.) P.Karst, Inonotus hispidus (Bull.) P.Karst and Ramaria formosa (Pers.) Quel. were evaluated for their antimicrobial activity against both gram positive (Bacillus subtilis and Staphylococcus aureus), gram negative (Escherichia coli, Proteus vulgaris, Klebsiella pneumoniae and Pseudomonas aeruginosa) and fungi (Saccharomyces cerevisiae Saccharomyces cerevisiae, Candida albicans, Penicillium chrysogenum and Aspergillus fumigates). The results revealed that ethyl acetate and methanolic extract of all the mushroom extracts showed significant antimicrobial activity against most of the bacterial and fungal microbes. However, the aqueous extract of these mushrooms was found either lacking or conferring insignificant antimicrobial activity. The ethyl acetate extracts of Ramaria formosa and Lentinus tigrinus produced more promising results against the bacterial microbes than fungal counterparts. Both ethyl acetate and methanolic extracts of Fomitopsis pinicola and Inonotus hispidus exhibited strong antimicrobial activity against the selected set of microbes. The antibacterial and antifungal activity exhibited by Fomitopsis pinicola at the concentration 150mg/ml was almost parallel to 10µg gentamycin and 50µg nystatin respectively. Therefore, Fomitopsis pinicola signifies as one of the promising mushroom species possessing strong antimicrobial activity against broad spectrum of microbes

Association of CYP3A5 Gene Polymorphism with Hypertension in Kashmiri Population

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Study of IRF-8 gene in Kashmiri population and its relation with Chronic Myeloid Leukemia

Chronic myelogenous leukemia (CML) is a clonal hematopoietic disorder caused by an acquired genetic defect in a pluripotent stem cell. A number of theories have been postulated to describe the etiology of CML such as genetic alterations and alterations in cytokine production. A combination of inflammatory cytokines have an important role in cancer development. The aim of this study was to screen for mutations of IRF-8 gene in CML cases and healthy controls of the Kashmiri population. We included eighty confirmed CML cases and an equal number of age, district and gender matched controls in this study. HaeIII enzyme digestion cuts amplified product at 5’-GGCC-3’ sequence and any mutation in it abrogates restriction digestion by this enzyme. Restriction results showed wild conditions with no mutation at any of the 6 positions where HaeIII cuts, which was confirmed by the sequencing results as well. Further sequencing results showed interesting single G A substitution at position 92 of the amplified product. In CML cases, the allelic frequency for normal allele (G) was found to be 47.5% (76/160) and the allelic frequency observed for G A type was found to be 52.5% (84/160). The allelic frequency observed in controls for normal allele (G) was 91.25% (146/160). The frequencies analyzed for G A allele was 8.75% (14/160). Since the frequency observed for G A allele was higher in CML cases (52.5%) than in normal controls (8.75%) and it was found to be statistically significant (OR= 11.52, 95%CI: (6.13-21.6); p = 0.001). Sequencing results further showed occasional deletion at the same position where transition was seen. Also we found that CML is prevalent more in males as compared to females (ratio is 1:1.12). Majority of the CML case were from district Srinagar of the Valley. Mean age of the cases and controls were found to be 44.7 years and 43.6 years respectively. We observed a higher representation of CML cases in the age group between 30 and 40. Furthermore, the presence of the Philadelphia chromosome (BCR-ABL fusion gene) was observed in 82.5% of CML cases in our study. This is the first report of the sequence variation in exon 7 region of IRF-8 gene and the risk to CML in the Kashmiri population. However, more insight need to be gained and further substantiated by conducting a similar study on statistically significant sample size

Polymorphism in MnSOD Gene and Breast Cancer Risk in Kashmiri Patients: a Case Control Study

Reactive oxygen species (ROS) have been implicated in the etiology of many human diseases. Antioxidant enzymes such as MnSOD protects cells from oxidative stress and generation of ROS. A case control study, with aim to evaluate the relationship between MnSOD Ala-9Val polymorphism and breast cancer was carried out. The study included 522 subjects, including 255 cases and 267 controls, 12 samples were missing or yielded no results. Genotyping of samples were carried out using PCR-RFLP method. We observed that neither of two conditions heterozygous (MnSOD Val/Ala) or Variant/(MnSOD Ala/Ala) was significantly associated with overall breast cancer risk. The frequencies of Val and Ala allele was almost similar in cases, however, a significant association was seen in case of older women (above 45 years of age) (OR =1.98, CI=1.07-3.66, P=0.04). Also ORs were elevated in case of women using oral contraceptives carrying Val/Ala genotype. (OR=2.20; CI=0.54-8.96). In conclusion MnSOD Ala-9Val polymorphism may modify the risk for breast cancer development particularly in presence of age above 45years, oral contraceptives, and urban life style

Genetic Variants at the Apo-A1 Gene in Association with Coronary Artery Disease

The aim of this study was to investigate whether the genetic variants of apolipoprotein A1: MspI polymorphisms in non translated region at -75bp upstream and +83bp in the first intron, had any impact on the development of CAD. A total of 400 unrelated adult subjects were enrolled in the study (200 CAD patients & 200 CAD-free controls) from the Kashmir region. Plasma levels of lipids were estimated for each sample by using photometric system. DNA extracted from blood samples was amplified by means of polymerase chain reaction, and then subjected to RFLP by using MspI enzyme; resolving the product on 3% agarose gel stained with ethidium bromide, and visualized under UV light. The G ?A substitution a

Isolation and characterization of a new cold-active protease from psychrotrophic bacteria of Western Himalayan glacial soil

Abstract As an approach to the exploration of cold-active enzymes, in this study, we isolated a cold-active protease produced by psychrotrophic bacteria from glacial soils of Thajwas Glacier, Himalayas. The isolated strain BO1, identified as Bacillus pumilus, grew well within a temperature range of 4–30 °C. After its qualitative and quantitative screening, the cold-active protease (Apr-BO1) was purified. The Apr-BO1 had a molecular mass of 38 kDa and showed maximum (37.02 U/mg) specific activity at 20 °C, with casein as substrate. It was stable and active between the temperature range of 5–35 °C and pH 6.0–12.0, with an optimum temperature of 20 °C at pH 9.0. The Apr-BO1 had low Km value of 1.0 mg/ml and V max 10.0 µmol/ml/min. Moreover, it displayed better tolerance to organic solvents, surfactants, metal ions and reducing agents than most alkaline proteases. The results exhibited that it effectively removed the stains even in a cold wash and could be considered a decent detergent additive. Furthermore, through protein modelling, the structure of this protease was generated from template, subtilisin E of Bacillus subtilis (PDB ID: 3WHI), and different methods checked its quality. For the first time, this study reported the protein sequence for psychrotrophic Apr-BO1 and brought forth its novelty among other cold-active proteases

Biochemical Characterization and Functional Analysis of Heat Stable High Potential Protease of <i>Bacillus amyloliquefaciens</i> Strain HM48 from Soils of Dachigam National Park in Kashmir Himalaya

A novel temperature stable alkaline protease yielding bacteria was isolated from the soils of Dachigam National Park, which is known to be inhabited by a wide variety of endemic plant and animal species of Western Himalaya. This high-potential protease producing isolate was characterized and identified as Bacillus amyloliquefaciens strain HM48 by morphological, Gram’s staining and biochemical techniques followed by molecular characterization using 16S rRNA approach. The extracellular protease of B. amyloliquefaciens HM48 was purified by precipitating with ammonium sulfate (80%), followed by dialysis and Gel filtration chromatography increasing its purity by 5.8-fold. The SDS–PAGE analysis of the purified enzyme confirmed a molecular weight of about ≈25 kDa. The enzyme displayed exceptional activity in a broad temperature range (10–90 °C) at pH 8.0, retaining its maximum at 70 °C, being the highest reported for this proteolytic Bacillus sp., with KM and Vmax of 11.71 mg/mL and 357.14 µmol/mL/min, respectively. The enzyme exhibited remarkable activity and stability against various metal ions, surfactants, oxidizing agent (H2O2), organic solvents and displayed outstanding compatibility with widely used detergents. This protease showed effective wash performance by exemplifying complete blood and egg-yolk stains removal at 70 °C and efficiently disintegrated chicken feathers making it of vital importance for laundry purpose and waste management. For functional analysis, protease gene amplification of strain HM48 yielded a nucleotide sequence of about 700 bp, which, when checked against the available sequences in NCBI, displayed similarity with subtilisin-like serine protease of B. amyloliquefaciens. The structure of this protease and its highest-priority substrate β-casein was generated through protein modeling. These protein models were validated through futuristic algorithms following which protein–protein (protease from HM48 and β-casein) docking was performed. The interaction profile of these proteins in the docked state with each other was also generated, shedding light on their finer details. Such attributes make this thermally stable protease novel and suitable for high-temperature industrial and environmental applications