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Home > Biochemical and Radio Labeling Studies of Venom Naja Naja Karachiensis With its Neutralization by Medicinal Plants of Pakistan

Biochemical and Radio Labeling Studies of Venom Naja Naja Karachiensis With its Neutralization by Medicinal Plants of Pakistan

Thesis Info

Access Option

External Link

Author

Hassan Bin Asad, Muhammad Hassham

Program

PhD

Institute

COMSATS University Islamabad

City

Abbottabad

Province

Punjab

Country

Pakistan

Thesis Completing Year

2016

Thesis Completion Status

Completed

Subject

Applied Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/7518/1/Muhammad_Hassham_Hassan_Bin_Asad_Pharmacy_2016_HSR_CIIT_20.09.2016.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725587818

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Biochemical and Radio Labeling Studies of Venom Naja naja karachiensis with its Neutralization by Medicinal Plants of Pakistan Background Snake bite envenomation is one of the vivid examples of neglected occupational hazards that accounts for tens of thousands of deaths all over the world. One of such instance is Naja naja karachisis bite, a nightmare for the inhibitants of Southern Punjab (Paksitan), often endup with countless deaths and sequela. To address this problem present study was designed to highlight scientific grounds for Naja naja karachisis envenomation and to rationalize folklore claimed Pakistani medicinal plants as a first aid treatment before proper hospitalization. Methods Proteomic characterization of Naja naja karchiensis venom was carried out with electrophoresis (SDS-PAGE) and HPLC (SEC & RP-HPLC) coupled LC-MS/MS whereas inorganic constituents were quantified with ICP-OES technique. Bio distribution and kinetic profile of venom was monitored with short lived radiotracer (99mTc) via direct radio isotopic binding technique. Lethal biological effects of crude venom were examined in terms of its LD50, hemolytic and anticoagulant behavior while toxic biochemical parameters (in vivo), towards liver (AST & ALT), heart (CK-MB & LDH) and kidneys (urea & creatinine) damage were investigated by following the recommendations of DGKC and IFCC methods. Venom was analyzed for different enzymatic activities (PLA2, ALPase, 5ʹ-ND, hyaluronidase and protease) by adopting conventional biochemical assays (in vitro). Twenty eight medicinal plants of Pakistan were extracted with methanol by simple maceration process and thereafter used to reverse deleterious actions of cobra venom. RP-HPLC coupled bioassay guided fractionation technique was used to characterize bioactive constituent/metabolite(s), responsible for anti-PLA2 activity in Bauhinia variegata L extract.
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شیخ انس یاسین

شیخ انس یٰسین
دوسرا حادثہ سعودی عرب کے سابق ہندوستانی سفیر شیخ انس یٰسین کی وفات کا ہے، ابھی وہ جوان تھے، لیکن تدبر و معاملہ فہمی میں تجربہ کار بوڑھوں سے کم نہ تھے، وہ مختلف اوقات میں مختلف ملکوں کی سفارت پر رہے، آج کل ٹرکی میں سفیر تھے، وہیں کار کے حادثہ میں وفات پائی، ان میں اپنے مذہب و ملت کا بڑا درد تھا، ہندوستان کے اسلامی اداروں سے ان کو خاص دلچسپی تھی، اپنی سفارت کے زمانہ میں متعدد اداروں کو دیکھا اور ان کی مدد بھی کی، دارالمصنفین کے بھی محسن تھے، یہاں آنے کا وعدہ بھی کیا تھا، مگر ایسے موقع پیش آتے رہے کہ آنا نہ ہوسکا، دو سال ہوئے دارالعلوم ندوۃ العلماء کے مدرسہ ثانویہ کی عمارت کا سنگ بنیاد رکھنے کے لئے لکھنو آئے تھے تو ان سے ملاقات ہوئی تھی، اﷲ تعالیٰ ان کی مغفرت فرمائے۔ (شاہ معین الدین ندوی،اگست ۱۹۷۴ء)

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Studies on the Production, Optimization, Partial Purification and Characterization of the Thermotolerant Carboxymethyl Cellulase from a Bacillus Strain Isolated from Local

CMCases are important enzymes that can hydrolyze lignocellulosic mass efficiently. Apart from this, CMCases are used in industries pertaining to textile, food processing, wine and brewery, pulp & paper, animal feed, agro based products, detergents, waste management, olive oil extraction and carotenoid extraction. The aim of this study is to explore the diversity of the indigenous thermophilic bacteria that are capable of degrading cellulosic biomass. A total of thirty-seven bacterial strains were isolated from stove ash samples. All the isolates were Gram positive and on the basis of physico-biochemical characterization identified and named as Bacillus sp TLW-1 to TLW-37. Plate screening was performed for ten different industrially important hydrolytic enzymes. Almost all the isolates were able to produce all the tested enzymes (except pectinase) with different profile. Positive CMCase producers were quantitatively screened by shake flask method. The isolate Bacillus sp. TLW-3 showed the highest CMCase production and was selected for further study. Isolate was identified on the basis of 16S rDNA sequencing as Bacillus licheniformis TLW-3. Sequence was submitted to NCBI (with KY440432 accession number). According to the growth curve, the isolate entered into log phase after 6 hrs of incubation and the log phase ended after 30 hrs incubation. Afterwards, the stationary phase began which ended after an incubation period of 72 hrs then the decline phase started. Highest enzyme production was found during the stationary phase. Optimization of production and growth conditions was performed by using two techniques i.e. OFAT and RSM. According to the OFAT approach, the highest growth and CMCase production were obtained in the medium containing 1% CMC, 1% peptone and 0.5% yeast extract with pH 7.0 at 50˚C and 150 rpm after the incubation period of 72 hrs. When compared with RSM optimization, the best growth was found at 70˚C in the medium having 0.5% peptone, 2.3% yeast extract and 2.52% CMC with pH 9.0 after 49.2 hrs of incubation. RSM optimized production conditions for highest CMCase production was found to be 1.8% peptone, 1.4% yeast extract and 1.6% CMC with pH 7.0 at 70˚C and 69.63 hrs of incubation period. Co-production of the enzyme was obtained in the medium containing 1% CMC and 1% starch as the sole carbon source. For the purification purpose, various precipitation and chromatography techniques were employed. CMCase was successfully (partially) purified with 8.250-fold and 13.75 % yield. Molecular weight of CMCase was estimated to be 28 kDa. Enzyme kinetics of partially purified CMCase demonstrated that enzyme was found active in the reaction mixture containing 1% CMC with pH 7.0 and at temperature 70˚C in the presence of Co+2, Ca+2 and Hg+2 ions. When the glucose was added in the reaction mixture enzyme activity was inhibited. The stability conditions of CMCase include, pH 6.0 and at temperature 4-50˚C; however, the stability was totally lost in the presence of the tested metal ions. The Vmax, Km and activation energy for CMCase were found to be 42.553 IU/ml/min, 9.587 mg/ml and 2.649 KJ/mole respectively. Endoglucanase gene was amplified, sequenced (11888 bp) and submitted to NCBI (with MF953225 accession number). Expasy-protparam online server was used for amino acid translation and in slicio physico-chemical characterization of the sequence. Secondary and tertiary structure of the protein revealed using SOPMA and ITASSER online server respectively.