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Isolation And Characterization Of Collagen Type Ii From Poultry Trachea

by Sidra Ashraf | Dr. Abu Saeed Hashmi | Dr. Sualeha Riffat | Zahid Mushtaq.

Material type: book Book; Format: print Publisher: 2011Dissertation note: This project was designed to use poultry waste to isolate and characterize collagen type II from its trachea. Collagen type II is being used along with condroitin sulfate and glucosamine for the treatment of osteoarthritis and is also available as a neutraceutical product in the market. For project purpose, trachea of slaughtered broiler birds were collected from the market and after removing adhering tissue and debris, it was then washed thoroughly first with distilled water and then with deionized water. Tracheal cartilage was then cut into small pieces and defattened with chloroform: methanol (2: 1 v/v) solution. After this, the cut pieces were properly cleaned with deionized water. 0.5% Pepsin solution in 0.5 M acetic acid was prepared. Cartilage was then hydrolyzed by the already prepared 0.5 % pepsin (in 0.5 M acetic acid) at 4 ° C for 48 hours. The extract was then separated from the tracheal pieces and the viscous solution obtained was centrifuged at 12000 rpm for 1 hr at 4 "c. Now the collagen was expected to be in the supernatant which was salted out by adding NaCI to a final concentration of 2.5M and kept for almost 12-16 hrs. This collagen was again centrifuged at 12000 rpm for 1 hr at 4 C. The obtained collagen pallet was redissolved in 0.5 M acetic acid and then it was dialyzed against 0.1 M acetic acid followed by dialysis with distilled water. The sample after dialysis was put in petri dishes and kept in freezer for overnight to let it be prepared for lyophilization. The frozen collagen sample was then lyophilized. After lyophilization, the sample gave an appearance of a white mesh. This sample was reconstituted in PBS with pH 8 to run it on SDS-PAGE. The procedure of SDS-PAGE in non reducing conditions was adopted for the characterization of collagen type II in the sample. The description of results of SDS-PAGE is given below: Lane M contains protein markers of different molecular weight. Lane 1, 2 and 3 contains samples at different steps of the whole procedure showing clear bands of collagen type II. Lane 4 contains lyophilized sample of collagen type II showing the thickest band (alpha chain of collagen type II). In this research, poultry waste has been used for making health improving product. As in our country poultry is used in bulk quantity so if its waste might be used in any medicinal product then it might not only be useful but also economical for such a developing country as ours. Another thing is that as this collagen Type II has been extracted from poultry trachea, it shows that tracheal cartilage is a rich source of such collagen type. Collagen Type II is used in the cure of arthritis especially rheumatoid arthritis so through this research, it has been made clear that poultry waste can be utilized in a positive way in medicinal industry and also that collagen Type II acts as an effective neutraceutical. Availability: Items available for loan: UVAS Library [Call number: 1330,T] (1).

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A Study On Prevalance Of Hcv Genotypes And Risk Factors Of Hepatitis C Virus In Punjab

by Tahira Tarar | Miss Faiza Masood | Dr. Muhammad | Mr. Zahid Mushtaq.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Hepatitis C virus (HCV) was marked as major agent which causes non-A, non-B hepatitis. Various partial and complete sequences of HCV nucleotide had been identified in the world. When these sequences were compared a marked genetic heterogeneity was revealed, that suggested the existence of HCV genotypes. Recent studies have pointed out the association of different HCV genotypes with different profiles of pathogenecity, infectivity and response to antiviral therapy. In our study we used typing system based on genotype specific primers focused on HCV 5´-UTR by using PCR. Genotype specific primers were designed for genotype 1, 2a, 2b, 3a, 3b and 4. 100 samples of HCV positive patients were collected. The frequency of occurrence of genotype 2a was 4 %, 2b was 5 %, 3a was 71 %, 3b was 11% and untypeable was 9 %. Blood transfusions, therapeutic injections, reuse of needles, dental procedures, shaving practices, body piercings, jaundice, dialysis, surgery and other multiple risk factors associated with HCV were studied. Major risk factor among females was therapeutic injections and among males the major risk factor was absence of shaving precautions. The study concludes that the most prevalent genotype in Punjab province is 3a. As each genotype sequence is different, the antiviral therapy against that particular genotype is also different. The treatment would only be successful if the genotype of HCV infected patient is known. This study will help in correlating efficacy of interferon therapy with different HCV genotypes and to understand the mode of transmission for hepatitis C virus. Availability: Items available for loan: UVAS Library [Call number: 1336,T] (1).

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Evaluation Of Pouifi As Candidate Gene To Have Influence On Milk Production In Red Sindhi And Dhani Cattle Through SNP Detection

by Waqas Tahir | Prof.Dr.Masroor Elahi Babar | Dr. Atif Hanif | Mr. Zahid Mushtaq.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2011Dissertation note: There are more than 200 million cattle in Pakistan which are a source of milk, meat, hide and drought purpose. Due to this large population of cattle, Pakistan is the fourth largest milk producer in the world. In spite of large number of cattle population in Pakistan, per animal milk production is yet quite low. That's why it is not wrong to say that there is still a great potential in the dairy sector of Pakistan to improve the overall milk production by improving the per animal Jilk production. This can be only achieved by improving the genetics of the dairy animals in Pakistan besides the other factors as well. Milk production is a polygenic trait. It is well established that growth hormone (GH) released from pituitary gland also plays an essential role' in growth, mammary gland development and lactation process. POUIFI factor is a member of the POU family of transcription factors that regulate animal growth and development and is also involved in the development of pituitary and regulation of hormonal expressions in animals. Therefore, genetic variation in the POU1Fl gene and its associations with growth traits in livestock animals could provide useful genetic markers for animal selection and breeding through marker-assisted selection (MAS). This study was conducted to genetically characterize !he POUIFI gene to identify the SNPs as genetic markers for future breeding and selection ~Ians. Keeping in view the importance of this gene all introns and axons were sequenced in smples of Red sindhi and Dhanni cattle to identify the SNP and validate the potential markers There are more than 200 million cattle in Pakistan which are a source of milk, meat, hide and drought purpose. Due to this large population of cattle, Pakistan is the fourth largest milk producer in the world. In spite of large number of cattle population in Pakistan, per animal milk production is yet quite low. That's why it is not wrong to say that there is still a great potential in the dairy sector of Pakistan to improve the overall milk production by improving the per animal Jilk production. This can be only achieved by improving the genetics of the dairy animals in Pakistan besides the other factors as well. Milk production is a polygenic trait. It is well established that growth hormone (GH) released from pituitary gland also plays an essential role' in growth, mammary gland development and lactation process. POUIFI factor is a member of the POU family of transcription factors that regulate animal growth and development and is also involved in the development of pituitary and regulation of hormonal expressions in animals. Therefore, genetic variation in the POU1Fl gene and its associations with growth traits in livestock animals could provide useful genetic markers for animal selection and breeding through marker-assisted selection (MAS). This study was conducted to genetically characterize !he POUIFI gene to identify the SNPs as genetic markers for future breeding and selection ~Ians. Keeping in view the importance of this gene all introns and axons were sequenced in smples of Red sindhi and Dhanni cattle to identify the SNP and validate the potential markers Availability: Items available for loan: UVAS Library [Call number: 1395,T] (1).

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