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51. Biochemical And Homology Analysis Of Jak2 Gene In Canines And Hominidae

by Marya Saadullah Khan (2014-VA-324) | Ms. Huma Mujahid | Dr. Abu Saeed Hashmi | Dr. Muhammad Yasir Zahoor.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Cancers are considered to be the most lethal of all diseases known out of which myeloproliferative neoplasms comprise of a very little percentage.The frequency of these disorders is known in human beings and a lot of work has been done on humans. But there is a lot of scope for research on this area in canines. As dogs were found to have strong homology with human beings, we compared canine cJAK2 exon 13 sequence with the humanhJAK2 exon 13 and found 96 % homology. Mutations in JAK2 gene are well known to cause three types of disorders i.e. polycythemia vera caused by a well-known point mutation in exon 14 causing substitution of valine for phenylalanine in JH2 domain of the protein.Essential thrombocythemia and idiopathic myelofibrosis may also be caused by this mutation but similar clinical conditions arise without the presence of this mutation. Studies have revealed that other point mutations such as deletion, addition or substitution are also responsible for these disorders. JAK2 is an intracellular protein which performs phosphorylation of STAT molecules upon their activation. Although the whole protein in its good state is important for its function but the two domains JH1 and JH2 are vital. JH1 domain acts as a tyrosine kinase enzyme and its activity is controlled by JH2 domain also known as pseudo tyrosine kinase domain. Any mutation in these domains leads to protein conformation defect and thus prevents its performance. Besides V617F mutation, other mutations are being discovered in this part of gene. Researchers have found mutations in exon 12, 13 and 15 that have been found to be involved in development of myeloproliferative neoplasms in different cases of patients. Blood picture do not reveal any direct clue except for increased erythrocytes alone or along with other cells like increased platelets. Therefore blood indices are not reliable parameter to indicate the type of mutation involved in these disorders. Also LDH and EPO levels are not correlated with the disorder. Although EPO test must be done to exclude the possibility of secondary PV and erythropoiesis. Availability: Items available for loan: UVAS Library [Call number: 2544-T] (1).

52. Molecular Exploration Of Zinc Finger Bed-Type Containing 6 Gene For Growth Trait In Beetal Goat

by Kanwal Rashid (2014-VA-496) | Dr. Maryam Javed | Dr. Asif Nadeem | Dr. Abu Saeed Hashmi.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Zinc finger, BED-type containing 6 (ZBED6), is a novel transcription factor.It acts as a repressor of IGF2 transcription in skeletal muscle myogenesis and development. it is mainly involved in organism development, signaling, cell to cell interaction, hepatic fibrosis, clathrin mediated endocytosis and tight junction signaling cascades. Chromatin immunoprecipitation (ChIP) sequencing using C2C12 cells identified about 2,500 ZBED6 binding sites in the genome, and the deduced consensus motif gave a perfect match with the established binding site in Igf2. Silencing of Zbed6 in myoblast cells affect Igf2 expression, cell proliferation, wound healing, and myotube formation. Genes associated with ZBED6 binding sites showed a highly significant enrichment for certain Gene Ontology classifications, including development and transcriptional regulation.Functional and signaling assays of BED6 gene indicate its probable role in controlling growth traits in Goat. Blood samples (n = 40) were collected. Inorganic method of DNA extraction used. Primers for PCR amplification will be designed using Primer3 software. PCR products will be sequenced bi-directionally on ABI 3130XL Genetic analyzer. The results of sequencing were analyzed using CHROMAS software. Sequence alignment tools (blast 2)were used for SNPs identification. Difference between allele and genotype frequency of studied gene evaluated by chi square test, likelihood test and analysis was done by POPGENE and one way ANOVA.Novel Variations identified which have probable implementation in selection of superior goats with higher tendencies towards weight gain. Availability: Items available for loan: UVAS Library [Call number: 2554-T] (1).

53. Sequence Analysis Of Mitochondrial Atpase 8/6 Gene Variants In Sindh Ibex (Capra Aegagrus Blythi)

by Javeria Zafar (2014-VA-222) | Dr. Asif Nadeem | Dr. Maryam Javed | Prof. Dr. Abu Saeed Hashmi.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: ATPase 8/6 gene plays a vital role in survival of an organism by generating energy in the form of ATP synthase. Considering the importance of ATPase8/6 gene in energy generating, present study has been designed to characterize this gene in Sindh ibex. The characterization of ATPase8/6 gene might be helpful for deriving phylogenetic relationship among different species and identifying new functions among the related species. Tissue/blood samples (n=15) were collected from Kirthar National Park, Sindh. Standard DNA extraction method was used for DNA extraction. PCR primers were designed by Primer3 software and amplification of gene was done by Polymerase Chain Reaction. PCR product was sequenced bi-directionally by Big Dye TM Terminator on ABI 3130XL Genetic Analyzer. Multiple sequence alignment was performed for polymorphism identification. Genetic diversity was calculated by using DNAsp. Phylogenetic analysis using the MEGA6 software package and an equally weighted maximum parsimony analysis was performed using the close-neighbor-interchange algorithm. The results indicated that Sindh ibex ATPase8/6 gene was highly similar to Capra caucasica. The results of this data might be helpful in designing effective conservation strategies of different species of wild animal. Availability: Items available for loan: UVAS Library [Call number: 2587-T] (1).

54. Effect Of Chicory (Cichorium Intybus L) Roots Powder On Blood Glucose Level Of Streptozotocin Induced Diabetic Rats

by Nazish Abid (2012-VA-782) | Dr. Sanaullah Iqbal | Ms. Tahreem Hussain | Dr. Abu Saeed Hashmi.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: The increasing numbers of elderly people, eating calorie rich diets, obesity and lack of physical activity have increased a great the number of patients with diabetes. Worldwide According to International Diabetes Federation (IDF) there were 6.9 million cases of diabetes in Pakistan in 2014 and occurrence of diabetes in adults of 20-79 years of age was 6.8 %.( International Diabetes Federation, cited on January 31, 2015). The feeding on chicory roots decreased the levels of plasma glucose, cholesterol, high density lipoprotein (HDL) cholesterol and also reduced liver cholesterol, triglyceride and total lipids of diabetic patient. In the present study, it was aimed to utilize a indigenous sources like prebiotics to overcome the representative burden on economy and estimating the outcome of administration of chicory powder for use as a substitute mediator of insulin in the control of diabetes mellitus. In the present study, chicory roots were cleaned, oven dried and ground to fine powder. The proximate analysis of chicory powder was performed. For feeding trial, thirty two Albino rats 5 to 6 weeks age, almost of same weight and mixed sex were procured and were randomly divided into four groups i.e. (A) Control , (B) Diabetic control, (C) Chicory treated and (D) Metformin treated group containing six rats each (three male and three female). Each group of rats was fed with a high fat diet (20%) for the first 2 weeks of adaptation. Then Diabetes was induced in B, C and D groups of rats by injecting 65mg/kg STZ through intraperitoneally. The diabetic rats of group C and D were then used for chicory intervention (125mg/kg of body weight) and for Metformin medication (500mg/kg body weight) along with normal diet respectively. The blood glucose level and weighing of animals measured initially and then Summary 39 weekly whereas glucose tolerance test of rats was performed initially and thereafter fortnightly. The feed and water consumption was measured on daily bases. A significant (p<0.05) difference in blood glucose level was seen among the group B, C and D of rats. A non-significance difference (p>0.05) in blood glucose level was observed when compared between male and female rats of groups A, B, C and D. A significant (p<0.05) difference in live body weight was observed. The group B, C and D showed significant (p<0.05) decrease in live body weight from day 7 to day 28 as compared to group A whereas a non-significant (p>0.05) difference in live body weight of male and female rats of all groups was observed. A significant (p<0.05) difference in glucose tolerance between group C and D was observed whereas a non-significant (p>0.05) difference of glucose tolerance was seen between all male and female rats of groups A, B, C and D while a significant difference (p>0.05) in feed consumption within group A, B, C and D was seen whereas a significant difference (p<0.05) in feed consumption was seen between the subjects. A significant gender difference in feed consumption was observed (p<0.05) between the subjects but within the subjects a non-significant difference was seen. A non-significant (p>0.05) difference was observed within the subjects in water intake whereas significant (p<0.05) difference was seen between the subjects whereas a significant gender difference (p<0.05) was seen between the subject but a non-significant gender difference was observed within the subjects. It is concluded that Cichoriun intybus root extract can be used as hypoglycemic agent to treat diabetic condition and has no adverse effects on body weight, feed intake and water intake. Availability: Items available for loan: UVAS Library [Call number: 2727-T] (1).

55. Comparative Genomic Study of Motor Neuron Disease in Horses and Human

by Shakeela Daud (2011-VA-534) | Dr. Muhammad Wasim | Prof. Dr. Tahir Yaqub | Dr. Abu Saeed Hashmi.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: cd not submitted Availability: Items available for loan: UVAS Library [Call number: 2810-T] (1).

56. Evaluation Of Bioactive Peptides/ Proteins/ Alkaloids From Extracts Of Croton Tiglium, Lawsonia Inermis And Eruca Sativa Against Mastitis Causing Bacterial Strains

by Rubia Saeed (2011-VA-377) | Dr. Muhammad Tayyab | Dr. Abu Saeed Hashmi | Dr. Muhammad Nawaz.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Mastitis is considered as one of the most prevalent disease in dairy animals of Pakistan. Bacteria which are found in most mastitis cases are S. aureus, S. agalactiae and E. coli. Infections caused by these bacteria are being treated by various antibiotics but due to their development of resistance towards these drugs, there is need to explore some alternatives like medicinal plant extracts for the treatment of mastitis. Croton tiglium, Eruca sativa and Lawsonia inermis have been reported to have antimicrobial activity, thus the extracts of these medicinal plants will be explored to their antimicrobial activity against mastitis causing bacterial strains. Present study purpose was to evaluate the bioactive proteins/alkaloids/peptides from extract of C. tiglium, E. sativa and L. inermis against mastitis causing bacterial strains. For this purpose, the leaves and seeds samples of selected medicinal plants (C. tiglium, E. sativa and L. inermis) were collected from Bagh-e-Jinnah and were identified from Department of Botany, University of the Punjab, Lahore. The ethanolic and aqueous extracts were prepared to evaluate the antimicrobial activity against mastitis causing bacterial strains. For this purpose, dust free leaves and seeds of selected plants were cut into small pieces, homogenized in ethanol/buffer and centrifuged. The resulting supernatant was then collected to check its antimicrobial activity against S. agalactiae, S. aureus and E.coli. Antimicrobial activity was analyzed by well diffusion method. Regarding the antimicrobial activity assay, the overnight grown cultures of the selected microbial strains was spread on the LB agar plates and the extracts was applied to wells incubated was done at 37°C for overnight. Inhibition zone was measured. Then the extracts having maximum activity were purified by GC.MS and the nature of extract was examined. All experiments were performed in triplicates so mean and average of the values was taken. Availability: Items available for loan: UVAS Library [Call number: 2853-T] (1).

57. Exploration Of Genetic Polymorphisms And Differential Expression Analysis Of Bovine Alpha-Lactalbumin And Osteopontin Genes Involved In Milk Composition

by Sidra Manzoor (2010-VA-92) | Dr. Asif Nadeem | Muhammad Imran | Dr. Abu Saeed Hashmi.

Material type: book Book Publisher: 2017Dissertation note: Economically important traits of dairy animals are usually controlled by a large number of genes. The identification of the single nucleotide polymorphisms in potential genes has been associated with economically important traits. During lactation, mammary epithelial cells produced large amounts of specific milk proteins. Due to the expression sites, physiological properties and chromosomal localization, LALBA and SPP1 genes might be considered as candidate genes for milk composition in buffalo. Alpha-lactalbumin (LALBA) gene has been reported to be highly transcribed in transition and peak phase while late lactation exhibited its decline with progressive rise in SPP1 expression. This project was designed to investigate the effect of single nucleotide polymorphism that influencing the gene expression thus modulates the milk protein content in Nili Ravi. Samples of unrelated Nili-Ravi buffalo were collected from two Government, Buffalo Research Institute, Pattoki, and Livestock Production and Research Institute (LPRI) Bahadarnagar Okara, livestock farms. Milk samples were collected at 15, 90 and 250 days lactation for expression analysis. The genomic DNA was extracted by using the standard Phenol Chloroform Isoamyl alcohol (PCI) protocol. Specific set of primers was designed for the amplification of the LALBA and SPP1 genes. The amplified PCR products were sequenced for the identification of SNPs. To determine the differential expression of bovine LALBA and SPP1 genes, RNA was isolated from milk samples using the TRIzol reagent and converted it into cDNA. Taqman probes were used that are specifically designed to detect and target the DNA sequence. Five intronic polym orphic sites were identified in LALBA while exonic regions exhibited a complete homology with reference sequence. Additionally, eleven polymorphisms were identified in bovine SPP1 gene, six were in coding region and five were Summary 122 found in intronic portion of the gene. The analysis and correlation of all identified polymorphism was done by using SNPs data analysis software “SNPator”. Results obtained from expression study was stored in in-build software of Real Time PCR and Cycle threshold (Ct) values of LALBA and SPP1 mRNA were compared in individuals of Nili-Ravi buffalo to determine the variation in expression levels. The LALBA gene expression was observed highest in transition phase with a gradual decrease of expression in mid and late lactation. The sample, NR-5, was observed highly expressed (79.30) while NR-2 with low expression (19.28) for alpha lactalbumin in early lactation. The change in LALBA regulation at same stage was considered due to genetic variation of the respective animal. While the SPP1 gene expression was observed with the highest values in peak lactation and remains elevated in late lactation. NR-4 has the highest (72.27) expression among all mastitis free healthy animals while NR-2 was observed with low expression. Thus, the identified SNPs might be used as genetic marker for milk production traits. Gene expression patterns may also help us to understand the molecular mechanisms of bovine LALBA and SPP1 genes influencing milk composition. However, the expression of both genes was considered in a correlation with other genes involved in milk production pathway. Also, the mutational effects of other milk proteins might be involved in determining the expression pattern of both genes in selected animals. Therefore, further studies are likely to explore the regulation of milk protein genes and their translational efficiency during the course of lactation in dairy animals. Availability: Items available for loan: UVAS Library [Call number: 2830-T] (1).

58. Production, Purification & Characterization Of Recombinant Thermostable Phytase And Its Biological Evaluation In Broiler Chicks

by Furqan Sabir (2007-VA-524) | Dr. Muhammad Tayyab | Dr. Abu Saeed Hashmi | Dr. Ali Raza Awan.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Phytate is the principle storage form of phosphorus in plants particularly in cereal grains and legumes. Mono-gastric animals doesn’t have ability to utilize phytate as phosphorus source. The animals release the undigested phytate from body with manure that cause environmental pollution. Phytases are responsible for the hydrolysis of phytate, resulting in availability of free phosphorus for the animal. The present study deals with the production and characterization of recombinant thermostable phytase and its biological evaluation in the broiler chicks. The PCR resulted in the amplification of 1.8 kb phytase gene using the genomic DNA of Thermotoga naphthophila as template. The purified PCR product was ligated in pTZ57R/T and the ligated material was utilized for the transformation of E.coli DH5α cells. The positive clones were selected on the basis of blue white screening. The restriction digestion of plasmid DNA from positive clones using NdeI and Hind III resulted in the release insert from the vector. The purified phytase gene after restriction digestion was ligated into pET21a already restricted with the same restriction enzymes and the expression was analyzed using E.coli BL21 CodonPlus (DEL) cells. SDS-PAGE demonstrated the intra-cellular production of recombinant phytase. The conditions were optimized for the optimal production of recombinant phytase (PHYTN). The maximal production of PHYTN was recorded when the BL21 CodonPlus cells having recombinant pET21a having phytase gene were induced with 1.4 mM IPTG and 6 hours post induction incubation period. The recombinant protein was purified using various chromatographic techniques and the purified protein was utilized for characterization. PHYTN showed optimal activity at 80 °C and pH 6 in sodium acetate buffer. The enzyme was found metal dependent and presence of Fe3+ or Cu2+ showed enhancing effect on PHYTN activity. Thermostability studies demonstrated that PHYTN retains 90% residual SUMMARY 71 activity when the protein was incubated at 80 °C for 1h in the presence of 1.5 mM Fe3+. The kinetic studies of PHYTN demonstrated km and Vmax values of 50 mM and 2500 μmole/min respectively when sodium phytate was used as substrate. The characterized PHYTN was used for poultry trials to check the efficacy of the enzyme in poultry birds. The results depicted that PHYTN put significant effect on the bird weight gain, feed intake and feed efficiency ratio. Presence of 1000 IU/kg of PHYTN resulted in the weight gain in 3rd, 4th and 5th week of trials from 504.766 to 533.535 g, 767.933 to 823.733 g and 999.833 to 1120.277 g respectively when compared with the control. The study demonstrated that this recombinant thermostable phytase is suitable for poultry feed industry and its domestic production will contribute the economic availability of PHYTN for the poultry feed industry. Availability: Items available for loan: UVAS Library [Call number: 2870-T] (1).

59. Effect Of Camel Milk Lactoferrin Against Carbon Tetrachloride Induced Hepatic Toxicity In Sprague Dawely Rats

by Nasreen Asghar (2014-VA-546) | Dr. Muhammad Nasir | Dr. Sanaullah Iqbal | Prof. Dr. Abu Saeed Hashmi.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Liver is a vital organ involved in regulation of several significant metabolic functions and is responsible for maintaining homeostasis of the body as well as detoxification of chemicals, drugs and other metabolites Chronic Hepatitis and mortality due to liver cirrhosis and hepatocellular carcinoma is common in Pakistan. Camel milk Lactoferrin has antiviral, anti-inflammatory, anti-carcinogenic properties. Liver cirrhosis is a serious and irreversible disease. it is common in Pakistan population which leads to mortality (Khan AA, 1995). (Anderson and Smith, 2001).These problems can be controlled by taking special measures. Nutraceutical foods like camel milk have many beneficial uses in this regard. Camel milk lactoferrin might be used for the cure of hepatic fibrosis induced by CCI4 in Sprague Dawely rats. Seventy five (75) male Sprague –Dawley rats were purchased from National Institute of Health Islamabad kept in animal house of UVAS Lahore and randomly divided into 5 groups under completely randomize design (CRD). In group (2-5) carbon tetrachloride (CC14) was subcutaneously injected with a mixture of 40% CCI4 (a mixture of pure CC14 and sterile olive oil) at 200 uL/100g body weight as single dose, 48 hrs before the starting of treatment or 0 day. After 48 hrs, rats were considered hepatic injured except the (+ve) control group. Among all groups, four groups –ve control, 30mg/kg/b.wt, 60mg/kg/b.wt and 90mg/kg/b.wt were supplied with standard diet plus Lactoferrin (in different concentrations and doses) orally, while control group were provided only standard diet throughout the efficacy study (30 days). Daily feed and water intake was monitored and cages were cleaned regularly. Body weight was recorded before decapitation throughout the experimental period. The efficacy studies data was analyzed through analysis of variance (ANOVA).Statistical significance was defined as P≤0.05.Means were compared for significance difference using Duncan‟s Multiple Range test (DMRt) and Least significance difference test (LSDt). Organ to body weight ratio and decreasing tendency in (-ve) control group was observed, while increasing tendency for body weight was observed in camel milk lactoferrin treatments groups during the study. In the present investigations, higher amount of transaminases (AST,ALT) and cholestatic liver enzyme (ALP) were observed however, their concentrations were significantly decreased significantly in camel milk lactoferrin treatments as compared to rising trend in (-ve) control group. Hypercholesterolemia is due to of oxidative stress induced by CCl4 and characterized by elevated levels of cholesterol .In the present research, it is concluded that camel milk lactoferrin treatments were effective in improving lipid is also obvious from the current results that camel milk lactoferrin treatments improved the hemoglobin (Hb) level although the effect was found non-significant .Likewise , results of present study also suggested that different treatments, time interval and their interaction had non-significant effects on white blood cells count in rats. Histopathology results of present study given exposed that CCl4 consequences prominent hepatic steatosis, hepatic cord rupture and necrosis .Post treatment of camel milk lactoferrin reduced the severity of CCl4 –induced liver intoxication. Fatty change and necrosis were improved in the histological sections of camel milk post-treated rats.(-ve) control group treated with CCl4 and basal diet showed severe hepatotoxicity, severe inflammation of hepatocytes and portal vein congestion, while group C and D treated with CCl4 ,basal diet and 30mg/kg/b.wt ,60 mg/kg/b.wt camel milk lactoferrin respectively showed moderate improvement in hepatotoxicity .group E treated with CCl4 and 90 mg/kg/b.wt lactoferrin showed mild hepatotoxicity in rats. Availability: Items available for loan: UVAS Library [Call number: 2943-T] (1).

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