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1. Analysis Of Medulla In Human Head Halr In Different Castes

by Summaiya Aurangzeb | Dr. Wasim Shehzad | Dr. Muhammad | Dr. Muhammad Tayyab.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1633,T] (1).

2. Molecular Characterization Of Cldn 14 Gene Encoding A Cell Tight Junction Protein In Mouse

by Ihsan Ullah | Dr. Muhammad Yasir zahoor | Dr. Wasim Shehzad | Ms. Shagufta.

Material type: book Book; Format: print Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1906,T] (1).

3. Identification Of Local Strain Of Toxoplasma Gondii Through Genotyping

by Saher Islam | Dr. Wasim shehzad | Mr..Akhtar ali | Prof. Dr Kamran ashraf.

Material type: book Book; Format: print Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1915,T] (1).

4. Optimization Of Nested-Pcr For Diagnosis Of Toxoplasma Gondii In Lahore Area

by Amna Arshad bajwa | Dr. Wasim Shehzad | Dr. Muhammad | Dr. Tanveer hussain.

Material type: book Book; Format: print Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1916,T] (1).

5. In-Silico Functional Prediction Analysis Of Prion Protein Polymorphisms In Sheep Scrapie

by Mubashar ahmad | Dr. Muhammad imran | Dr. Muhammad | Dr. Wasim shehzad.

Material type: book Book; Format: print Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2012,T] (1).

6. Application Of Microsatellite Markers For Genetic Diversity Analysis Of Endangered Punjab Urial (Ovis Orientalis Punjabiensis) In Pakistan

by Anam Aftab (2012-VA-534) | Dr.Tanveer Hussian | Dr. Wasim Shehzad | Dr. Muhammad Tayyab .

Material type: book Book; Literary form: not fiction Publisher: 2014Dissertation note: Biological diversity is now recognized as common concern of mankind and genetic diversity is the major driver of variation within and across breeds, which helps populations to adapt to environmental changes. There is very little importance is given to conserve wild sheep in previous years and its genetic diversity is decreasing day by day. Every now and then breeds are being haunted for crossing to some other imported breed without attempting to see if such efforts will be sustainable. For any breed development efforts thus, available genetic resources need to be characterized both at phenotypic and genetic levels (Khan et al. 2007). Among the three levels of Biodiversity, one is Genetic variation which is suggested bythe International Union forNatureconservation (IUCN) for preservation (Mc Neely et al. 1990). The reason for it is that firstly; genetic diversity favors the changes as the environment changes and secondly; it prevents inbreeding depression (Reed and Frankham 2003). In this way genetic diversity increase the survival status and increase fitness of individuals. Among many other wild animals present in Pakistan, there are 6 to 9 species of wild sheep (Ovis orientalis) are present which have different color and size of their winter neck ruff of males, saddle patches and horns color. Urial is a picture of Marco Polo in texture and hue. In Pakistan, ladakh urial, Blanford urialand Punjab urial are found in Gilget, Baluchistan and Punjab respectively. The discrepancy lies in the color of ruff among these 3 sub species (Roberts, 1977). Urial is among those precious wild animals that were hunted severely for trophy and other purposes in the past, that’s why included in red list of IUCN in vulnerable category (IUCN 2000). Punjab Urial (A type of wild sheep – Ovis vignei punjabiensis) belongs to family bovidae which is the large family consisting of 140 species (Glazko et al. 2011) is facing serious threat of extinction in Pakistan is a medium-sized wild sheep which is included in IUCN red list of endangered animals (IUCN 2002). Urial is inhabitant in Western Central Asianregion stretching from northeast side of Iran and west side of Kazakhstan to Balochistan (Pakistan) and Ladakh regions of North India. The local name of Urial is Shapo, Arkar and Gad. Reddish-brown outstretched pelt that achromatizes during the winter is one of the distinct traits of urial (Aleem 1977; Schaller 1977). Urial is gregarious and sexually dimorphic as males are called rams and females as eves (Awan 2001). Male have weightof 40 kgand have large spiralled horns having height of 80 to 100 cm and females have comparatively less weight and height of 25 kg and 12 cm respectively and have uncurled horns.Females give birth to 1 or 2lambs in recent days of April (Awan, 2001). Males have a black ruff expanded from the neck to the trunk and notably longer horns. Table 1.1:Some physical features of the Punjab Urial (Awan et al. 2001) Features Urial Body weight 40 kg male; 25 kg female Shoulder size 31-35 inches or 80-90 cm Horn size 39 inches or 80-100 cm long male; 12 cm long female Urial are found in moderate to very arid habitats, especially grasslands including agricultural fields and woodland areas (Valdez, 1982). Urial is herbivorous and eats grasses, shrubs and grains. The patch of salt range of Pakistan which fall in the area of Pind Dadan Khan, Choa Saidan Shah and Kallar Kahar is considered like a paradise on earth for the wild fauna. The fascinating hills of these areas are covered with thick trees and different wild plants are also the sanctuary of urial. Table 1.2: The details of these sub species in IUCN list of endangered mammals (IUCN 2002). Subspecies Citation in IUCN list Ovis vegnei blanfordi VUC 1 Appendix ll Ovis vegnei punjabienses ENA1cde,c1+2a Appendix ll Ovis vegnei vegnei VUC 1 Appendix ll In Pakistan, Punjab Urial dispersed throughout the Kala Chitta and Salt Range in a very little number(Hess et al, 1997). The Afghan Urial inhabits Baluchistan, Khyber Pakhtunkhwa, and Sindh Provinces. In Chitral District, little segregated populations of Ladakh urial are stillextensively distributed near the west bank of the Kunar river from Chitral southwards to Drosh. Ladakh Urial existence at the east bank of Kunar river and north of Chitral are not proved (Malik 1987). Total population estimate of urial is recorded by conducting several surveys. Reasonably 2,500 to 3,000 urial existsin Baluchistan (Hess et al. 1997). In 1993 the overall population assessment of Northern Areas was four hundred to five hundred Urial (Rasool 1999). In Pakistan, there were seemingly less than 600 Ladakh urial (Hess et al. 1997;NWFP 1992; Schaller 1971, 1977) but the number of urial decreased to 200 to 300 urial in all over northern areas (Rasool 1999). Based on the facts mentioned above there is dire need to conserve the Urial population in the country. The urial is one the precious fauna of Pakistan and provides us with wool and meat (fat, flesh or any eatable part). It is also important in economic way and in maintaining of ecosystem balance. In the beginning, sheep were reared for meat, milk and skin (Ensminger and parker, 1986). After 3500 B.C. men learnt to spin wool and so used wool in textile industry (Smith et al. 1997). So, because of increasing world population, there is great demand of these products and increasing day by day. That’s why we have to conserve is natural resource of Pakistan.Habitat fragmentationleads to the risk of exaggerated genetic drift and inbreeding in isolated population. So, there is need to save urial from these threats by enforcement of law and conserving it for future. In all over the euchromatic genome Microsatellite markers are present. These markers are highly polymorphic (Ellegren 2000; Schlotterer 2000).A lots of polymorphic microsatellites have been analyzed in ruminants like domesticsheep, cattle etc. (de Gortari et al. 1997,1998 ;Hayes et al.1996; Jenkings et al. 1997) aiding the use of these in parentage testing.   Microsatellite markers are among the most reliable molecular markers for genetic characterization studies in animal species (Sunnucks, 2001) and are simple sequence repeats (SSRs) of 1-6 base pairs, repeated tandemly in coding as well as noncoding portion of DNA in prokaryotes and eukaryotes (Weber and May, 1989; Toth et al., 2000). Microsatellite markers have often used for genetic diversity studies because they areabundant, unbiased, widely distribution all over the DNA, highly polymorphic, easy in assessment like genotyping of these markers (Canon etal. 2001).Microsatellite markers aid in genetic differentiation and conservation studies (Peter et al. 2007;Rendo et al. 2004; Arranz et al. 2001).These are considered very useful markers for assessment of genetic diversity, parentage confirmation, genome mapping, disease research population genetic studies and conservation genetics. These are also reported to be efficient enough to identify within and among breed differentiation and population sub structuring in cattle (Glowatzki-Mullis et al. 1995; Ciampolini et al. 1995; Garcia-Moreno et al. 1996; Jarne and Lagoda, 1996; MacHugh et al. 1998).Therefore the conservation activities are very important to save Punjab Urial from extinction and the study is designed to explore its genetic diversity. Availability: Items available for loan: UVAS Library [Call number: 2261-T] (1).

7. Microbiological Quality Assessment Of Human And Veterinary Drugs

by Sadaf Riaz (2007-va-277) | Dr. Ali Ahmad Sheikh | Dr. Fareeha Akhtar | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: The continued increase in population results in increasing demand for pharmaceutical products. According to Pakistan pharmaceutical manufacturers association, Pakistan has approx. 400 pharmaceutical industries including 25 multinational industries. The Pakistan pharmaceutical industry meets 70% of country’s demand of pharmaceutical drugs. Medicines are chemical compound that are administrated to human or animal as an aid to diagnosis, treatment or prevention of disease (Lecca, 1978). Microbiological quality assessment is very important point of pharmaceutical manufacturing. The term quality in its wide sense means “Safety “. Microbial contamination means, presence of undesired micro-organisms or their metabolic products (Uba, 1990). Pharmaceutical drugs are used in many ways in treatment of diseases (Mugoyela et al. 2010). Pharmaceuticals of different forms are susceptible to contamination by different microbes (Aulton, 2002). Contamination of medicines with micro-organisms can bring about changes in their physiochemical properties, results in product degradation before their expiry date (Shaikh et al. 2000). Microbial contamination rang from true pathogen like Clostridium tetani, to opportunistic pathogens, such as Pseudomonas aeruginosa (Mwambete, 2009). Microbial infections not only due to physical presence of micro-organisms, but also their metabolites/toxins can be very harmful even in low quantities (Shukla et al. 2004). Medicines used in treatments cannot afford to have microbial contamination. Patients who take medicines have very weak immune system which makes them vulnerable to wide range of infections. Microbial contamination of drugs may contribute to secondary microbial infections in immunologically weak patients (Adeshina et al. 2009). Contaminated pharmaceuticals may lead to medication complicacy in patients. Drugs can be divided into two types on the basis of microbiology; sterile pharmaceutical products and non-sterile pharmaceutical products (Clement et al. 2013). Sterile pharmaceutical products should not contain any viable micro-organisms and non-sterile pharmaceutical products may contain viable micro-organisms but it should be within official limit and all drugs should not have any pathogenic bacteria. Most of drugs can be contaminated during storage and handling (Takon and Antai, 2006). Antimicrobial agents are added to medicines to minimize microbial growth but should not be added to mask poor manufacturing process (Gad et al. 2011). The level of microbial contamination in medicines depends on availability of nutrients, water, presence of other micro-organisms, osmotic pressure and oxygen etc. The factors determining the results of drug-borne infections may include the type and amount of microbes, patient’s immune system and route of administration (Baird, 2004). In recent years manufacturing of drugs have improved. The occurrence of microbes in drugs has been well documented. There have been many reports of infections caused by contaminated drugs (Ibezim, 2002; Coker, 2005). But majority of cases of infections caused by medicines are not recognized and reported as such (Denyer and Baird, 2006). Previous studies have showed microbiological quality concern related to commercially available drugs (Denyer and Baird, 2006). Contamination of drugs results in loss of faith of a consumer in pharmaceutical industry and sales would go down (Mugoyela and Mwambete, 2010). Although pharmaceutical industries are one of the growing and expanding sector in Pakistan, but the quality of medicines varies as they are retail oriented. Unfortunate situation is present in sales of drugs. A number of unlicensed drugs stores selling poorly manufactured drugs 2 Hence the microbiological quality of pharmaceuticals, assessment of number and type of bacteria and fungi within drugs, is essential to ensure consumer safety (Urmi et al. 2014; Tamalli et al. 2013) This study to relate to national need and reflects its importance. New international trend focus on the quality of pharmaceutical products, unfortunately few studies has been carried out in this prospective which is not proven to be sufficient in predicting the quality of drugs. This has compelled me to do research on this topic in order to assess the level of such microbial contaminants in pharmaceuticals given to patients in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2275-T] (1).

8. Antibiotic Resistance Pattern Of Clostridium Perfringens Type A From Poultry And Its Resistance Modulation Using Medicinal Plant Extracts

by Arifa Jabeen (2008-VA-398) | Dr. Ali Ahmad Sheikh | Prof. Dr.Aftab Ahmad Anjum | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Avian acute necrotic enteritis (NE) is the burning issue in poultry sectors worldwide resulting in huge economic losses. Cl. perfringens; causative of NE; is an anaerobic, rod shaped Gram positive, spore former, catalase negative bacteria which produce characteristic double zone of hemolysis on 5 % sheep blood agar. A total of 100 (n=100) intestinal samples were collected from ten different poultry farms in and around Lahore. Intestinal contents were enriched in fluid thioglycollate medium (FTM), isolated and purified on the tryptose sulfite cycloserine (TSC) agar which is highly selective and recommended medium for Cl. perfringens; identified by microscopic examination and typical biochemical tests (catalase and blood hemolysis). Results showed that fifty six out of 100 samples were positive for the presence of Cl. perfringens with the overall positivity ratio of 56 % that is the highest percent prevalence in broilers and type was confirmed on the detection of cpa alpha toxin producing gene resulting in the product band size of 128 bp through PCR. Five PCR confirmed isolates were subjected to antibiotic resistance studies. According to the results and CLSI standards, ampicillin and amoxycillin were susceptible to Cl. Perfringens while tetraacycline, ciprofloxacin and streptomycin were found resistant to all of the five isolates. By agar well diffusion method plant extracts (Astragalus, Zingiber officinale, Calotropis procera and Gymnema sylvestre) were tested for their anti-clostridial activity. All produced zones of inhibitions of varying diameters except the aqueous extracts of 1st three plants that Summary 75 produced no any zone of inhibition on agar plates against any of the five isolates tested while those of Gymnema sylvestre produced good zones. MIC of plant extracts were determined against isolates. The extracts of Gymnema showed the lowest MIC values among all and in between sequential extracts of plants, the chloroform extracts showed relatively low MIC values in comparison to others statistically using One way ANOVA. MIC values of plant extracts were determined in combination with the specific MIC breakpoint concentrations of antibiotics too for their resistance modulatory effects against the isolates. Statistical result findings were very effective in combination with the resistant antibiotics as MIC values significantly reduced in comparison to performed solely with plant extracts. Especially the modulatory results for ethanol and chloroform extracts of nearly all four plants were noticeable as MIC values were rapidly declined below 100 in comparison to plant extracts alone where MIC values were higher to 500. All of the results data obtained through whole experiment were analyzed using SPSS versions 20.0. Hence on the basis of above findings with the current study, it is concluded that medicinal plant extracts may prove effective herbal therapeutic agents against Cl. perfringens type A; the potent cause of necrotic enteritis in poultry. The findings of the study might be helpful in renewing and rescheduling the antibiotic administration plan with the use of medicinal plant extracts to modulate the action of antibiotics. Availability: Items available for loan: UVAS Library [Call number: 2276-T] (1).

9. Seroprevalence And Molecular Detection Of Brucellosis In Hospitalized Patients With Clinical Manifestations Of Brucellosis

by Riffat Yousaf (2008-VA-342) | Dr. Wasim Shehzad | Prof. Dr.Tahir Yaqub | Dr. Haroon Akbar.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Brucella species are host specific facultative intracellular pathogens which cause brucellosis in both animals and humans. Brucellosis is one of the most common zoonotic diseases worldwide. In Pakistan, the incidence of brucellosis is increasing day by day due to lack of awareness of this deadly malady. It is transmitted from infected animals to humans who are in close contact with infected vaginal secretions, feces, blood, aborted fetus, or by consumption of unpasteurized milk and dairy products. Infection due to B. melitensis and B. abortus are mostly prevalent for brucellosis in human. Total 218 blood samples were collected in gel vacutainer tubes from hospitalized patients who were clinically manifested with brucellosis. Out of 218, 12 RBPT positive blood samples were collected in EDTA containing vacutainer tubes separately. Serum was isolated from all blood samples (without EDTA). These serum samples were first screened by Rose Bengal Plate Test (RBPT). DNA was extracted from all positive RBPT blood and serum samples and randomly selected negative RBPT serum samples. All extracted DNA (≤10ng/µL) were subjected to Brucella genus and two species specific (B. abortus and B. melitensis) Quantitative Real Time Polymerase Chain Reaction (qRT-PCR) assay. Furthermore, few selected extracted DNA (≥20ng/µL) from blood and serum samples were examined by genus and Multiplex specie specific PCR. The PCR products were electrophoresed on 2.5% agarose gel. Then selected products were sequenced by ABI 3130 XL sequencer. The data were analyzed by SPSS software using Chi square test. The present study helped to diagnose accurately and precisely brucellosis in clinical manifested patients, which is further helpful for devising the strategies to control this disease. Availability: Items available for loan: UVAS Library [Call number: 2286-T] (1).

10. Comparitive Immunogenic Evaluation Of Purified And Whole Culture Vaccine Of Pasteurella Multocida B2 Of Bovine Origin

by Anika Khalid (2006-VA-357) | Dr. Imran Altaf | Dr. Jawad Nazir | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Pasteurella multocida is responsible for causing Heamorrhagic Septiceimia which is a fatal and acute disease mainly associated with bovines. Pasteurella multocida consists of two important antigenic structures i-e Outer membrane protein(OMP) and Lipopolysaccharide(LPS) that forms capsule.These two structures played vital role in pathogenicity as well as in the immunogenicity. In our present project we study the role of LPS,OMP alone and in combination as an immunogens.We compared the immunogenic behaviour of whole culture vaccine(consists of LPS and OMP) purified vaccine(consists of OMP),LPS vaccine (consists of LPS only) by IHA and CFT (using OMP and LPS both as an Ag separately) and the overall results showed that whole culture HS Alum precipitated vaccine produced better antiboby titer against both outer membrane proteins (OMP) and lipopolysacharrides (LPS) when checked against respective antigens as compared to the vaccines containing OMP and LPS alone. So it was concluded that LPS no doubt act as an immunogen but its immunogenicity increases many times when combine with protein (OMP) as in whole culture vaccine. Availability: Items available for loan: UVAS Library [Call number: 2316-T] (1).

11. Genetic Association Study Of Apolipoprotein A-V (Apoa5) And Sortilin (Sort1) Genes With Risk Of Coronary Artery Disease

by Irfan Basharat (2012-VA-802) | Dr. Akhtar Ali | Dr. Wasim Shehzad | Dr. Muhammad Tayyab.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: In developed countries cardiovascular disorders are prominent cause of death. One third deaths in the world are due to cardiovascular disorders. Among cardiovascular disorders coronary artery disease responsible for one in five deaths in USA. Its main reason is the lipids values particularly cholesterol and triglycerides in the blood. An estimation made by WHO indicated that 9 million people die per year due to hypercholesterolemia. 100 blood samples were collected from patients of coronary artery disease and from normal patients with no myocardial history. Allele specific primers for SORT1 gene and APOA5 genes were designed using Primer 3 software web facility. Genomic DNA will be amplified by PCR then genotyping will be carried out and DNA will also be sequenced. Hardy-Weinberg principle and Fisher Exact test were used to assess the allele frequency and significant variations from results When patient of MI and normal group were genotyped and sequenced we find out that there are 34 AA homozygous, 1 GG homozygous and 12 heterozygous persons in case of APOA5. The SORT1 person shows 24 GG homozygous and 3 AA homozygous and 13 heterozygous persons. Our study shows a definite association between APOA5 and SORT1 with respect to MI disease persons. This study shows a significant association of single nucleotide polymorphism in APOA5 and SORT1 genes with coronary artery disease. Availability: Items available for loan: UVAS Library [Call number: 2326-T] (1).

12. Expression And Purification Of A Potent Surface Antigen (Sag1) Of Toxoplasma Gondii In Prokaryotic Expression System

by Zunaira Zafar (2009-VA-542) | Dr. Wasim Shehzad | Dr. M. Yasir Zahoor | Dr. Imran Rashid.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Toxoplasma gondii, an intracellular obligate parasite infects almost all warm-blooded animals including human. Toxoplasmosis, caused by T. gondii, may show minute to severe clinical results in humans. Currently, there is no vaccine available for human use. SAG1 is a major candidate of interest for vaccine development that elicits humoral as well as cellular immune response against this devastating parasite. rSAG1 that had already been ligated in pET28/His expression vector, was transformed in E. coli (BL21) host and expression was confirmed by means of SDS-PAGE and western blotting. Nickel columns were utilized for affinity based chromatographic purification of rSAG1. This purified protein was then quantified via protein quantification kit. Immunogenic recombinant SAG1 can be used in diagnostic antigen-antibody tests e.g. in ELISA. Moreover, it might be used in vaccination against T. gondii. Vaccine against this parasite may alleviate socio-economic burden on human society that ultimately modulates the health parameters for better living. Availability: Items available for loan: UVAS Library [Call number: 2393-T] (1).

13. Mutational Analysis Of Hcv Gene Encoding E1 Glycoprotein

by Muhammad Saad Junjua (2013-VA-893) | Dr. Muhammad Imran | Dr. Wasim Shehzad | Dr. Abu Saeed Hashmi.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Hepatitis C virus (HCV) is a positive single stranded RNA virus that belongs to Flaviviridae family and causes liver related issues like hepatocellular carcinoma, cirrhosis and chronic liver disease. HCV is affecting people worldwide; more than 170 million peoples have been affected so far and the number is increasing day by day. Its prevalence in Pakistan is about 3% to 6%. There is lot of variation in its genome and it is classified into 6 major genotypes and these genotypes are further classified into many subtypes. Size of HCV is about 9500 bps which only encodes single polyprotein. This 3000 to 3300 amino acids polyprotein is processed by cellular and viral proteases to generate 10 polypeptides consisting of 4 structural (Core, E1, E2 and P7) and 6 non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A, NS5B). The most immunogenic gene from all the genes is E1. It involves in the interaction with the host cell and easily escapes from the immune system of host due to the presence of hypervariable regions in E1 gene. To isolate the E1 gene, RNA extraction was done using the kit method. RNA was converted to cDNA which was then amplified in two rounds of PCR using nested primers from HCV core region. After confirming the presence of HCV RNA in serum samples, PCR amplification of HCV E1 gene was carried out using gene specific nested primers. Amplified E1 gene products were Sanger sequenced and aligned with standard sequence to find out genetic variations. E1 gene sequences were converted to protein sequences for which secondary protein structures were made and analyzed. No noticeable change was seen in these secondary protein structures. The protein sequences were also analyzed for the presence of B-cell and T-cell epitopes; two T-cell epitopes (QAFTFRPRR, FLVGQAFTF) were found which may inform the development of a proper vaccine against HCV. Availability: Items available for loan: UVAS Library [Call number: 2399-T] (1).

14. Molecular Diagnosis Of Anaplasmosis In Buffaloes

by Muhammad Salman (2008-VA-135) | Prof. Dr. Khalid Saeed | Dr. Muhammad Imran Rashid | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Bovine Anaplasmosis is a tick-borne haemo-rickettsail disease, caused by Anaplasma species transmitted mechanically by flies, biologically by ticks and blood contaminant fomites. It is an economically important tick-borne disease of buffalo in tropical and sub-tropical areas of the world. In current study, we developed and optimized PCR first for detecting Anaplasma at genus level in buffaloes. One hundred (100) blood samples were collected from buffaloes around the Lahore region. The stained thin blood films were examined microscopically and 37% blood samples were found positive for intra-erythrocytic bodies which were then selected for DNA extraction. The DNA was extracted using commercially available kit for eventual use in optimization of PCR for diagnosis of bovine Anaplasmosis. The primers were designed targeting 16S rRNA gene of Anaplasma. For the detection, the PCR product was run in 2% agarose gel stained with ethidium bromide and thirty seven samples showed the amplification band at 179bp. The selected samples were sent for ABI sequencing to Singapore for the accurate detection of the Anaplasma species. The sequencing results were blasted with database of Genbank and we observed homology with Anaplasma phagocytophilum. We found 37% prevalence of Anaplasmosis in buffaloes through PCR. However more studies are required to confirm the species of Anaplasma infecting buffaloes (Bobalus bobalis) by designing species specific primers. Furthermore, additional studies are needed to establish the epidemiology of Anaplasmosis by using molecular tools in different geographical areas of the country for their better control. Availability: Items available for loan: UVAS Library [Call number: 2389-T] (1).

15. Sero-Screening Of Camels For Different Infectious Diseases

by Mazia Khalid (2008-VA-358) | Dr. Aamir Ghafoor | Prof. Dr. Aftab Ahmed Anjum | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Camel is the precious and important animal in Pakistan. Camel is the most well adapted livestock species, survives and produces in climatic extremes and is well appreciated for its significance in the pastoral economy of the province. The camel being an important livestock species uniquely adapted to hot and arid environments and therefore contributes significantly to the food security of the nomadic pastoral households. Although camel being hardiest animal is less susceptible to diseases as compared to other livestock animals but literature shows that some diseases are still prevalent in camels. In view of the significance of camel as livestock animal as well as the symbol of cultural heritage of the nomadic pastoralists, there is a need to combat different diseases to which camels are susceptible and then appropriate control strategies should be applied. Present study was designed to check the percentage positivity of different major diseases in camels that may pose serious issue relating to camel health and its importance as an important livestock animal. The diseases included in this study are Q fever, Brucellosis, FMD, CBPP and Neosporosis. ELISA is used to detect antibody prevalence by using specific kit based protocol for each disease whereas in case of Brucellosis RBPT is also used as basic screening test. And it was found that Q fever has highest percentage seropositivity in both districts as compared to other diseases whose presence in camels was found to be almost seronegative. So it was concluded that camel is still resistant to many diseases though some diseases are still prevalent in camels and these diseases should be controlled through public awareness and routine screening. Availability: Items available for loan: UVAS Library [Call number: 2401-T] (1).

16. Indigenous Elisa Kit For Toxoplasma Gondii: Optimization Of Antibody Detection Elisa Of Sag 1 Protein As An Antigen In Mouse Model

by Madiha Sana (2013-VA-957) | Dr. Muhammad Imran Rashid | Dr. Haroon Akbar | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Toxoplasma is an apicomplexan intracellular parasite which is the cause of toxoplasmosis in man and animals. It occurs by the ingestion of oocyst from feces of cats or by eating raw meat in which cysts are present. It is the one of the major cause of encephalitis and abortion in immuno-compromised animals and humans. As it is difficult to screen out infected live animals from field, it is important to vaccine animals as well as humans for toxoplasma to prevent its transmission from animals to humans and from humans to their off springs. Cloning of surface antigen genes plays an important role in development of vaccine and for serology of T. gondii. Enzyme linked immuno-sorbant assay proves to be a significant tool to estimate the humoral response elicited against expressed recombinant protein in mice. The recombinant protein of SAG1 was collected from Molecular Parasitology Laboratory, University of Veterinary and Animal Sciences, Lahore. In the previous studies, SAG1 sequence was cloned in the expression plasmid and successfully expressed in prokaryotic expression system. In the current study, rSAG1 was quantified by using BCA protein assay through BioWORLD protein quantification kit. In another experiment, the Swiss mice were immunized with 15 μg rSAG1 protein 3 times with 2 weeks intervals. Two groups of mice were formed with five mice in each group. Sera were collected after 2 weeks of each inoculation. For performing ELISA, four different experiments were performed with different concentrations i.e. 5μg/ml, 250μg/ml and 500μg/ml with two different dilutions; 1/50 and 1/20. The O.D. values of concentrations 5μg/ml and 250 μg/ml with two dilution series of 1/20 and1/50 were not observed significant while the antigen coating concentration of 500 μg/ml with 1/50 dilution showed 1:160 titre and with 1/20 dilution showed 1: 1280 titre after the 3rd shot. The O.D values with 500 CHAPTER 6 SUMMARY SUMMARY 36 μg/ml concentration with 1/20 dilution after the 3rd shot were observed significant in the inoculated group as compared to the O.D values of un-inoculated negative group. It is suggested to carry out ELISA with purified rSAG-1 protein and to optimize ELISA to test toxoplasma infected mice. Availability: Items available for loan: UVAS Library [Call number: 2433-T] (1).

17. Characterization And Thermostability Of Phytase Produced By Indigenous Aspergillus Niger Isolates

by Madeeha Tariq (2010-VA-293) | Prof. Dr. Aftab Ahmad Anjum | Dr. Jawad Nazir | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Phytase enzyme now becomes more important commercially. Presence of phytate in food and feed make them less nutritive due to phytate complexes mainly with mineral ions and proteins. Phytase in monogastric animals and human stomach either produced in small amount or not. This leads to phosphorous Pi deficiency. Supplementation of food and feed with phytase enzyme full fill this deficiency through degradation of phytate complexes and release of Pi. Degradation of phytate complexes makes phosphorous other mineral ions and amino acids available for growth and development. It was proved that feed conversion rate in poultry increased due to supplementation of phytase in poultry feed. Feed of monogastric animals mostly at industrial level pelleted to give it a shape or to kill microorganisms (sterility). At industrial level enzyme production and processing cost about 2 billion. So this demands a thermostable phytase to use at industrial level or its cost effective production. Aspergillus niger have been used industrially for production of beneficial enzymes. A. niger isolates procured from department of microbiology were confirmed through macro and microscopic characteristics as A. niger. These isolate were screened for phytase production on phytase screening medium PSM agar. Positive isolates identified through noval staining using 2% cobalt chloride, 6.25% ammonium molybdate and 0.42% ammonium vanadate for contrast. Positive isolates next proceeded for phytase enzyme production in broth media (pH 5.6) using 0.5% sodium phytate as substrate. Incubation was done at 30oC for 5-7 days in shaking incubator 150rpm. After production quantification of enzyme was carried out through enzyme activity assay. There maximum (274.99±10.14 FTU/ml) and minimum (68.88±2.55 FTU/mL) activity of phytases from isolate PASN01 and PASN08 was observed. Phytases characterized through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) to know protein molecular weights. Highest molecular weight 107.82kDa was PASN06 and lowest was 35.21kDa of PASN 01. Aspergillus niger spores subjected to steam heat treatment at 30oC, 45oC, 60oC, 75oC and 90oC for 15, 30, 45, 60minutes to identify thermostability. At 30oC and 45oC temperature, spores of A. niger isolates found to be thermostable. But at 60oC, 75oC, or 90oC treatment spores become inactivated or there 6.0 logarithmic reduction in spore count was observed. Thermostability of phytases was found at 60oC, 75oC, 90oC for 15, 30, 45, and 60 minutes treatments. Enzyme from A. niger PASN01 and PASN08 observed as thermostable at 60, 75 and 90oC. Phytases from PASN01 and PASN08 showed 160.55±42.96 and 00±.00 FTU/mL decreased in activity after 45 minutes of treatment at 60oC temperature, respectively. PASN01 phytase displayed 163.88±23.35, 172.77±7.52 and 171.66±7.26 FTU/mL decreased in activity after 60minutes treatment at 60, 75 and 90oC. In case of PASN08 phytase at 60, 75 and 90oC temperature after 60minutes treatment, 13.33±10.41, 16.66±6.00 and 23.88±41.37 FTU/mL decreased in activities were observed, respectively. PASN08 phytase observed more thermostable than other phytases of A. niger isolates. Enzyme can bear pelleting and pre pelleting temperatures. Enzyme from PASN08 also observed stable during storage at room temperature. Conclusion: A. niger PASN08 spores inactivated or killed and phytase observed stable at 60oC temperature, after 60mins treatment. Temperature 60oC may be used industrially for cost effective thermostable phytase production from indigenous A. niger isolate PASN08. Availability: Items available for loan: UVAS Library [Call number: 2475-T] (1).

18. Role Of Maoa Polymorphism In Criminal Violence Among Convicted Offenders

by Shahpal Shujat (2010-VA-494) | Dr. Maryam Javed | Dr. Wasim Shehzad | Dr. Saadat Ali.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Criminal violence is the violent act of crime that can be of different category such as Homicidal, Sexual and Physical violence. Criminal violent behaviour is under the control of brain having serotonergic and dopaminergic system that is under the influence of genes .MAOA gene lies under the control of serotonergic system. There are 2 polymorphic forms of MAOA gene on the basis of its activity i-e MAOA-L (Low activity allele) and MAOA-H (High activity allele). MAOA-L in males but in females MAOA-H is linked with violent behaviour in combination with environmental factors. Blood/Saliva /Buccal swabs samples (n = 20) were collected from District Jail Sheikhupura, Pakistan Organic/Inorganic method of DNA extraction was used. Primers for PCR amplification was designed using Primer3 software. PCR products were sequenced by using Sanger method from CAMB. Violent behaviour was determined by using Buss and Perry aggression Questionare Physical aggression scoring. Sequencing results were analyzed using CHROMAS software. Sequence alignment tool like CLUSTAL was used for aligning multiple sequences of convicted samples and compared with control (DNA mixture amplification) and standard samples and detected for SNP. Then SNP were detected for the amino acid change by using ExPASy software.SNP statistical analysis was done by calculating POPGENE software and association analysis was performed by using ANOVA. SNP in exon 8 at locus 43591035 of MAOA was identified that was T instead of G while in exon 13 two SNPs were identified at locus 43603089 and 43603112 .Both SNPs for exon 13 was heterozygous and changes T into A .The synymous SNPs were at locus 43591035 and Summary 73 43603089 . But the SNP at locus 43603112 was non-synonymous .The association study showed that there was no association between SNP and violence scores among convicted offenders. Availability: Items available for loan: UVAS Library [Call number: 2486-T] (1).

19. Antibiotic Resistance Profiling Of Enterococcus Faecium Recovered From Retail Chicken Meat From Lahore City

by Amina Habib (2010-VA-313) | Dr. Ali Ahmad Shiekh | Dr. Fareeha Akhtar | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Enterococcus faecium is gram positive bacteria which are normal intestinal flora of warm blooded animals and humans beings. It is responsible for various types of diseases such as neonatal meningitis, endocarditis, urinary tract infections and intra-abdominal or pelvic wounds. Retail chicken could be a source of Enterococcus faecium. Irrational use of antibiotics in chicken rearing can lead to emergence of antibiotic resistant Enterococcus faecium. The chicken meat gets contaminated at the time of slaughtering and resistant bacteria may transfer to human beings through food chain. In present study prevalence of E. faecium recovered from retail chicken meat samples collected from various areas of Lahore city was estimated. A total 43 chicken sample (leg or wing) were processed for isolation of E. faecium. Identification of E. faecium was made using standard culturing and biochemical reactions. Out of 43 samples, 30 samples (69%) were found positive for Enterococcus faecium. Antibiotics resistance profiling showed that the isolates were resistant to following antibiotics mentioned as below: Ampicillin (100%) >Tetracycline (73%) > Erythromycin (53%) > Ciprofloxacin (46%) > Chloramphenicol (40%) > Rifampicin and Vancomycin (36%) > Teicoplanin (33%) > Doxycycline (20%) > Fosfomycin (0%). From the study it is concluded that retail chicken is the carrier of antibiotic resistant Enterococcus faecium and could transfer resistance to humans. Efforts should be made to use antibiotics wisely and hygienic practices should be followed during slaughtering and processing of chicken meat to avoid bacterial spread from animal source to human beings. Availability: Items available for loan: UVAS Library [Call number: 2489-T] (1).

20. Isolation And Antibiotic Resistance Profiling Of Enterococcus Faecium Recovered From Retail Fish In Lahore City

by Maria Butt (2010-Va-281) | Dr. Ali Ahmad Sheikh | Prof. Dr. Aftab Ahmad Anjum | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Enterococcus faecium is an enteric, gram positive and lactic acid bacteria which belongs to genus enterococcus and inhabit the intestinal tract of human, fish and other warm blooded animals. Due to irrational use of antibiotics in human and veterinary sector, antibiotic resistance has been developed in commensal bacteria including Enterococcal species. These resistant bacteria are released in environment through human and animal waste and transfer resistant genes to susceptible bacteria present in wetlands making them antibiotic resistant. E. faecium is considered to be involved in transmission of resistance genes, present on mobile genetic elements through conjugation to other bacteria. The resistant bacteria can be transferred to human through food chain. The present study was designed to evaluate the prevalence of E. faecium recovered from retail fish samples collected from various areas of Lahore city. Antibiotic resistance profiling of the isolates against commonly used antibiotics was also determined. In current study 65 fish samples (intestinal swabs) were processed for isolation of E. faecium through standard culturing and biochemical reactions. Out of 65 swab samples, 30 samples (47.69%) were found positive for Enterococcus faecium. Antibiotics resistance profiling showed that the isolates were resistant to antibiotics mentioned as below: Ampicillin (100%) > erythromycin (56.6%) > rifampicin (53.3%) > Chloramphenicol (30%), ciprofloxacin (30%) > tetracycline (20%), vancomycin (20%) > Teicoplanin (13.3%) > Doxycyclin (6.6%) > Fosfomycin (0%). E. faecium isolates showed resistant to at least 2 or 3 antibiotics of different group. In conclusion it is observed that retail fish is the carrier of antibiotic resistant Enterococcus faecium and Summary 51 could transfer resistant genes to wetlands and other aquaculture from where it could be transferred to human body. Efforts should be made to use antibiotics wisely and hygienic practices should be followed during slaughtering and processing of fish meat to avoid bacterial spread from animal source to human beings. Availability: Items available for loan: UVAS Library [Call number: 2493-T] (1).

21. Studies On Biological Control Of Salmonellosis In Poultry

by Kiran Imtiaz (2010-VA-296) | Dr. Ali Ahmad Sheikh | Prof. Dr. Masood Rabbani | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Contaminated poultry food products are the main cause of Salmonellosis all over the world. Salmonella may enter in different poultry farms through vertical or horizontal transmission. Previously, Salmonella infection and number were controlled in poultry and their products by using different methods such as usage of hazard analysis critical control point (HACCP), antibiotics, symbiotics, etc. But not any method gives the better result that shows the reduction of Salmonella species in poultry that cause infection. Therefore, the attention was diverted to find the alternative therapies such as the usage of bacteriophage, Bacteriophage specific to Salmonella used in reduction of Salmonella number as a biocontrol agent in poultry origin. It proves effective for Salmonella reduction. In the current study samples for isolation of Salmonella and bacteriophages were collected. Salmonella was isolated from liver, caeca and lungs of infected chickens collected from different infected poultry farms. After processing of sample according to the literature method, confirmation of salmonella from samples were done by inoculating them separately on S.S agar by streaking method. Further confirmation of Salmonella was done on the basis of biochemical testing. Bacteriophage was isolated from sewage water near to the poultry farms. For their isolation, sewage sample was centrifuged and then supernatant was collected in separate tube. Filtration was done of Supernatant, and then Filtrate was used as a bacteriophage source. Bacteriophages in filtrate were confirmed by spot test method. After confirmation of both, in vivo study was performed to evaluate the effect of bacteriophage on Salmonella when inoculated in combination or separately in chickens groups. 51 Summary Statistical analysis: The data will be transferred on spreadsheet using MS Excel 2010. The data will be analyzed through one way ANOVA test using Statistical Package for Social Sciences (SPSS) version 18.0. The present study was helpful to determine the effect of bacteriophage in reduction of Salmonella in commercial broilers sector. Antibiotic resistant Salmonella infection is difficult to control using conventional ways of antibiotic therapy and is responsible for huge economic losses in poultry. Chicken groups that were used for invivo study showed that bacteriophage was proved very effective in reduction of Salmonella either gives it in combination with Salmonella or separately to poultry. It is predicted that bacteriophage therapy is better than the conventional ways for reduction of Salmonella infection in poultry sector. It is essential that the research should be continue to study the effect of bacteriophage in reduction of specie specific Salmonella, and to determine the effect of different physicochemical factor on their activity. Availability: Items available for loan: UVAS Library [Call number: 2511-T] (1).

22. Development Of Dna Based Diagnosis Of Theileriosis In Cattle And Its Specificity With Blood Smear Microscopy

by Uzma Sarwar (2014-VA-777) | Dr. Wasim Shehzad | Dr. M. Yasir Zahoor | Dr. Ali Ahmed Sheikh.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Theileria annulata and Theileria parva are intra-erythrocytic parasites which are responsible for causing tropical theileriosis and East Coast fever in cattle respectively. This parasite is transmitted by ticks to vertebrate host i.e. cattle. Currently used diagnostic methods for diagnosis of bovine theileriosis are clinical symptoms, peripheral blood smear microscopy and serological tests (IFAT and ELISA). Current study was conducted to compare the specificity and sensitivity of blood smear microscopy and PCR techniques to diagnose bovine theileriosis. This study is comparative as well as developmental in nature. Although peripheral blood smears microscopy is cost effective and quick method of diagnosis in case of high or moderate parasitaemia in blood. But the limitations associated with microscopy include false negative diagnosis in case of low parasitaemia in chronic and asymptomatic infection, morphological similarity of Theileria with other species of Plasmodium and Babesia. These limitations may lead to misdiagnose the infection due to which disease may remain unnoticed. PCR based method, developed in this study, and is found to be more specific and sensitive than conventional microscopy. Fifty blood samples were collected from September, 2015 to November, 2015. These samples were screened microscopically as well as with PCR for presence of Theileria. Nine samples were found to be positive microscopically but 18 samples were found positive by PCR. The results obtained from the study clearly show that PCR is more reliable, precise and sensitive assay for diagnosis of bovine theileriosis then microscopy. It is hoped that proposed method to diagnose Theileria will help to nullify the problems associated with microscopy. This will ultimately facilitate in the formulation of effective treatment control and vaccine development strategies. Availability: Items available for loan: UVAS Library [Call number: 2547-T] (1).

23. Identification Of Genetic Variants In The Low Density Lipoprotein Receptor Gene Causing Familial Hypercholesterolemia And Its Sequence Homology With Mus Musculus

by Ameer Hassan (2014-VA-504) | Dr. Wasim Shehzad | Dr. Muhammad Yasir Zahoor | Dr. Muhammad Tayyab.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Familial hypercholesterolemia (FH) is caused by mutations in the genes coding for the low-density lipoprotein receptor (LDLR), apolipoprotein B-100, or proprotein convertase subtilisin/kexin type 9 (PCSK9). In this study, a molecular analysis of LDLR gene was performed in 20 unrelated patients from Pakistan. All patients were clinically diagnosed with definite or possible hypercholesterolemia according to a uniform protocol and internationally accepted WHO criteria. Preferable study was made to highlight the Genetic variation in Exon 4 of LDLR gene associated with defective catabolism of cholesterol effecting lipid metabolism which results in Familial Hypercholesterolemia. The extraction of genomic DNA was done from all selected blood samples. By selecting primers they were synthesized and optimized on extracted DNA samples. PCR product was sequenced and aligned. Mutations in the LDLR gene and its sequenced homology with Mus musculus were analyzed. We didn’t found any polymorphisms in the LDLR gene exon 4. So we concluded that there is no association between SNPs and increased levels of cholesterol in Pakistani population. More research should be carried out in Pakistan by increasing the sample size and considering the other regions of LDLR gene. This study will help the early detection and treatment of such cases and may ultimately reduce the incidence of mortality due to myocardial infarction. Apart from diagnosis, we also suggest it will be a potential therapeutic strategy to manage FH. Availability: Items available for loan: UVAS Library [Call number: 2538-T] (1).

24. Assessment Ofgenetic Polymorphism In The Tph Gene As Susceptible Factor For Aggressive Behavior In Criminals From Prisonsof Punjab, Pakistan

by Zonash Riaz (2010-VA-479) | Dr. Saadat Ali | Dr. Muhammad Yasir Zahoor | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Aggression is perceived as hostile, injurious, or destructive behavior often caused by frustration, can be collective or individual. Genetic studies have associated several genes with aggression in humans. One of the candidate genes that turned out to be associated with aggression, anger, and impulsivity is the tryptophan hydroxylase (TPH) gene. We investigated the polymorphism in the TPH gene in the unrelated male individuals in the Punjab ethnic backgrounds who were administered the Punjabi translation of Buss and Perry aggression questionnaire. The questionnaire measured four aspects of aggression: physical aggression, verbal aggression, anger and hostility (Buss and Perry, 1992).Scores ± SD of 83.544± 26.63 was obtained for Buss and Perry aggression questionnaire. TPH is a rate-limiting biosynthetic enzyme in the serotonin pathway and regulates levels of 5-hydroxytryptophan (5-HT) by converting tryptophan into 5-hydroxytryptophan, which is the direct precursor of 5-HT. It is conceivable that variations in the TPH gene could contribute to low activity of the 5-HT system. Single nucleotide polymorphisms (SNPs) that show associations to aggression and anger-related traits have been detected in intron 7 of TPH gene.DNA of individuals categorized into controls and criminal groups was extracted by organic method of DNA extraction.The targeted region of the TPH gene was amplified by the primers designed against intron seven. The amplified Pcr product was precipitated and it was sent for sequencing. The resultant sequenced data was then compared on the basis of Buss and Perry aggression scores. All unrelated male individuals from the Punjab ethnic groups were assessed on the scales showing scores for physical aggression, verbal aggression, anger and hostility. The minimum score for the respondents were 65 and highest score for the respondents were 135 among the criminal group while control have minimum scores of 50 and maximum scores of 113.Mean scores and standard deviations were calculated for criminals and control groups. Control group havephysical aggressionmean scores ± SD19.318 ± 6.21, verbal aggressionmean scores ± SD17.590± 4.41,angermean scores ± SD23 ± 6.868and hostility mean scores ± SD23.636± 9.12and total mean scores ± SD83.544± 26.63while criminals have physical aggressionmean scores ± SD28.2±8.134, verbal aggressionmean scores ± SD20.4±4.427, anger mean scores ± SD27.3±6.97and hostilitymean scores ± SD28.1±7.72and totalmean scores ± SD04.2±20.47.Mean aggression values for the criminals was 104 and for controls was 83, higher in criminals as expected. Criminals groups exhibited greater level of aggression as compared to that of control groups on the basis of four scales of aggression i.e. physical aggression, verbal aggression, anger and hostility. Observed genotypic frequencies among the control groups were 0.7 for CC, 0.3 for the AC and 0 for AA whereas genotypic frequencies amongst criminal group were 0.3 for CC, 0.6 for AC and 0.1 for AA. Controls carried higher genotypic frequencies for normal CC genotype than criminals whereas the genotypic frequencies for AA and AC genotypes were higher in Criminal group.Observed allelic frequencies amongst the control group was 0.8 for C and 0.15 for A whereas observed allelic frequencies amongst the criminal group was 0.4 for A and 0.6 for C. Controls carried higher allelic frequencies for the normal C allele while criminals carried higher allelic frequencies for A allele.In our study proportion of the less common (A or U) alleles was 40%, and the proportion of the more common (C or L) alleles was 60% in criminal group as compared to 15% of A allele and 85% of C allele in the control group. Statistical analysis has associated significantly Criminals and controls group at P value less than 0.05. Advances in the understanding of the genes modulating aggression can contribute meaningfully to a rational assessment and treatment of individuals with pathological aggression and a predisposition to violence. Results can be utilized for the screening of Aggression in the individuals for forensic applications. In future studies, other polymorphism in TPH and other aggression related genes may also be analysed in Pakistani population. Availability: Items available for loan: UVAS Library [Call number: 2595-T] (1).

25. Molecular Characterization Of Oca2 Gene In Correlation With Eye Color For Forensic Application

by Anam Noor (2014-VA-942) | Dr. Muhammad Yasir Zahoor | Dr. Allah Rakha | Dr. Saadat Ali | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: DNA phenotyping is the use of genetic information such as DNA to determine a phenotype. It helps forensic investigator to predict the physical appearance of an individual to find unknown perpetrators or to identify missing persons using molecular analyses from biological samples in cases where all other means of inquiry, including conventional DNA profiling are non-informative. In a non-forensic setting, it permits the prediction of the physical appearance of our ancestors, historical persons or any other deceased individual for whom the identification of appearance traits may be interesting, and it sheds light on human evolution. Based on current research there are only a few traits for which it is possible to make an accurate description based on underlying genetic variation. Eye color is a complex polygenic trait and is under the control of many genes. There are infinite number of eye colors with a multitude of patterns and mixtures. Almost 74% human eye color is under the control of OCA2 gene on chromosome 15. This gene correlates with the physical appearance of eye color as EVCs (externally visible characteristics) therefore it can be used as a parameter in forensic application. Samples collected from local areas of Pakistan is divided into two groups brown that include samples from 17 individuals and other than brown including 15 individuals. DNA of 32 samples was extracted and samples were amplified against a selected sequence of OCA2 gene containing SNP rs1800407, which was previously reported to be associated with eye color in European populations. These amplicons were sequenced using Sanger sequencing and chromatograms obtained were analyzed by pairwise and multiple alignment tools. The results show the presence of5 polymorphic sitesin various samples including SNPsrs1800407 and rs1900758. These polymorphic sites were further analyzed by applying t-test which shows no significant association between retrieved polymorphic sites and eye color.The results show no significantly associated marker with eye color to be present within the selected sequence so we need to analyze other markers or SNPs which could be found to be associated with eye color that would be very useful in forensics application. Availability: Items available for loan: UVAS Library [Call number: 2623-T] (1).

26. Microbiome Analysis Of Human Normal Specific Flora From Skin Of Laborers And Academic Professionals Of Lahore For Forensic Application

by Talha Umair (2014-VA-941) | Dr. Wasim Shehzad | Dr. Saadat Ali | Dr. Muhammad Yasir Zahoor.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Human microbiota or normal flora is the aggregate of microorganisms that resides on the surface of skin, oral mucosa, conjunctiva and GIT. Human skin has a complex variety of microbial system and varieties of microbes mean that they are potential source of forensic identification because human microbiome varies individual to individual due to differences in hygiene, professions and region to region because of some environmental factors and microbial flora can shed more frequently upon touching any kind of surfaces and microbes are left for long time at any surface so can be identified easily. Human microbiota varies individual to individual so it may become potential source for forensic identification of individuals through specific microbiome analysis. Fourty Samples were obtained by swabbing from the palm surfaces of hands and soles of feet of individuals of different professional groups in order to recover bacterial communities. Bacterial culturing and Bacterial DNA extraction followed by the implementation of 16S rRNA amplification by polymerase chain reaction (PCR) and sequencing of the PCR product, allowed an even more comprehensive broad range investigation of bacterial communities. Bioinformatics analysis was done to compare microbial communities. This research elaborated the significance of skin microbial communities in identifying individuals and can be a major contribution in forensic science to find and identify individuals when there is less major evidence, i.e. human DNA and body fluids. Availability: Items available for loan: UVAS Library [Call number: 2639-T] (1).

27. Development Of Novel MtDNA Metabarcodes For Species Differentiation Of Class Reptilia

by Imran Tariq (2014-VA-505) | Dr. Muhammad Imran | Dr. Wasim Shehzad | Dr. Saadat Ali.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: The FoImer COI: mtDNA. universal primers that are considered standard for DNA barcoding of life contain so many =matches against the target sequences of vertebrate men tat they often end in failure to amplify many of vertebrate DNA eurections. This disaepancy fawn foe the seaman and designing of new metabarcode panes that can be used m ideally all inditdrals of vertebrees or at least all individuals represented in a class of Vertebrate such as Cass Reprilia. The current study was embadang on such an endeavor The proposed study was develop new m5DNA membarc ode that may be used as universal Kilns; to amplify almost all species of Class Repalia for different formic and molectdr biodivesity analyses. Blood and tissue samples were collected from Class Repdha (at :east 24 species from every ceder reported to be present in Pakistan) DNA was extracted from the collected specimen through stacdasd organic method. qualified and =meted and then PCR-amplified using novel universal primers selected from aligned =DNA sequences origtadng from all repdlian mitochondria DNA pnomes submitted to diens online sequence databases such as NCB: micleotide database. Tne sensitiviry. of PCR was assessed using a range of DNA come:madam. The amplified products were sequenced on A131 Genetic Analyzed following Sarge's dideacy method of sequencing. The correctness of obtained croDNA sequences were examined visually in Chromes Lite 2.1 software and then alipmmt of these sequences were per: waxed agitinc highly similar DNA sequences in NCBI nu6eonde databases using BLAST in order to identify the coigin of la-noun =DNA sequences sequencing everimeas and phyla...net< studies was confirm the specificity of the universal primer set developed and present a novel metabarcode for species level identification of large number of reptelian species. So, In future this barcode can be used for species identification in various fields of study such as illegal trade and molecular estimation of boidiversity. Availability: Items available for loan: UVAS Library [Call number: 2628-T] (1).

28. Homology & Polymorphism Analysis Of Cc2d1a Gene In Human And Canine For Cognitive Function

by Hafiz Qamar Abbas (2014-VA-214) | Dr. Muhammad Yasir Zahoor | Dr. Wasim Shehzad | Dr. Saadat Ali.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Cognitive disability is a group of genetically heterogeneous abnormality that leads to variable degrees of cognition deficits. It has been shown that inherited disorders can be caused by mutations in large number of different genes and there is evidence for the presence of as yet unknown genes in a significant proportion of patients. This disease can affect 1-3% of overall population and higher in consanguineous families. We aimed to identifying the homology and polymorphism of the gene CC2D1A between human and canines. The present research work was carried out in four phases. The first phase was including enrolment of 10 affected non relevant families with disease history and consent was taken on consent forms as approved by IRB, UVAS. Secondly DNA extraction was done by using standard lab protocols. Thirdly amplification of the selected domains of selected gene (CC2D1A) was done through PCR amplification after designing primers of the selected domains. Sequencing of the amplified products has to be done through Sanger method and mutation analysis was conducted for variants We found two new asynonymous mutation one is deletion of c. 1664_1664delA which lead to the change in the normal function of protein (88%) and other is heterozygous mutation c.1921A/T that result in amino acid change from R to W (12%). Whereas homology analysis shows that deletion region is partially conserved as it code different amino acid but some key domains are conserved. This homology shows that deletion in this region can change the protein expression which can relate to unconscious condition like behavioral or mental retardation. This will be helpful in providing genetic counseling services to indigenous population for intellectual disability cases. Availability: Items available for loan: UVAS Library [Call number: 2627-T] (1).

29. Molecular Characterization Of Canine Babesiosis In Ticks And Dogs

by Tahira Sarwar (2014-VA-523) | Dr. Wasim Shehzad | Dr. M. Yasir Zahoor | Dr. Ali Ahmed Sheikh.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Babesia canis is an intra-erythrocytic parasite which cause canine babesiosis in both animals and humans. Currently, there are three sub-species of Babesia canis has been identified i.e Babesia canis canis , Babesia canis vogeli and Babesia canis rossi. Currently used diagnostic methods are clinical symptoms, peripheral blood smear microscopy and serological tests (IFAT and ELISA).Current study was conducted to compare the specificity and sensitivity of blood smear microscopy and PCR techniques to diagnose canine babesiosis. This study is comparative as well as developmental in nature. Although peripheral blood smear microscopy is cost effective and quick method of diagnosis in case of high or moderate parasitaemia in blood. But the limitations associated with microscopy include false negative diagnosis in case of low parasitaemia in chronic and asymptomatic infection,morphological similarity of Babesia with other species of Plasmodium and Theileria these limitations may lead to misdiagnose the infection due to which disease may remain unnoticed.Total 50 samples comprising of 25 blood samples and 25 ticks were collected randomly from infected dogs from June, 2015 to November, 2015. These samples were screened microscopically as well as with PCR. Out of 50 samples of dogs and ticks, 18 samples found to be positive for the Babesia canis. 11 samples are Babesia canis vogeli and 07 samples are Babesia canis canis were to be identified in positive samples of dogs and ticks.The results obtained from the study clearly show that PCR is more reliable, precise and sensitive assay for diagnosis of canine babesiosis then microscopy. It is hoped that proposed method to diagnose babesiosis will help to nullify the problems associated with microscopy. This will ultimately facilitate in the formulation of effective treatment control and vaccine development strategies which may eradicate babesiosis. Availability: Items available for loan: UVAS Library [Call number: 2642-T] (1).

30. Anthelmintic Activity Of Withania Coagulans Against Gastrointestinal Nematode Of Sheep In District Killa Saifullah, Baluchistan

by Yousaf Gul (2009-VA-145) | Dr. Muhammad Lateef | Dr. Saadullah Jan | Dr. Muhammad Imran Rashid | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Evaluation of anthelmintic activity of Withania coagulans was studied against GIT nematodes of sheep in district Killa Saifullah Baluchistan. Sheep of the district were screen out for the presence of GIT nematodes. Animal positive for GIT nematodes and having 150+ Egg per Gram (EPG) of feces was included in the drug trial. Animals were treated with extract(s) of locally available herbal plant (withania coagulans) and levamisole. Two types of plant formulations that is crude powder and crude methanole extract were prepared each with various dosages. The effect of both medicinal plant and levamisole was observed on different groups of animals and the results were analyzed with appropriate statistical tool. Eighty animals were randomly divided in to eight groups (10 animals in each group) i.e. A, B1, B2, B3, C1, C2, C3 and D. Animals in group A served as control untreated group. Animals in groups B1, B2 and B3 were treated with crude powder of Withania coagulans at the dose rate of 1, 2 and 3 g/kg body weight respectively. And Animals in groups C1, C2 and C3 were treated with crude methanol extract of Withania coagulans at 33.3, 66.6 and 100mg/kg equivalent dose rate of 1, 2 and 3 g/kg body weight respectively. Animals in group D were given Levamisole at the standard dose rate of 7.5 mg/ kg body weight. Data was analyzed by using SPSS version 20.0; comparative analysis was done by applying ANOVA. P value <0.05 was taken as significant. The analyzed data and the results revealed that Levamisole is still a better anthelmintic against ovine nematodes in district Killa Saifullah Balochistan. Efficacy of levamisole tested for 15 days in-vivo sheep was up to 92%. This efficacy was much higher than the various forms and dosages of medicinal plant. The efficacy of Levamisole was significantly higher (P<0.05) than all forms and dosages of medicinal plant. Group C3 treated with crude methanol extract of Withania coagulans at the dose rate of 10mg/kg equivalent to 3mg/kg showed highest efficacy of the plant that is up to 48%. The efficacy showed by the form of the medicinal plant used in group C3 against ovine GIT nematodes was significantly higher (P<0.05) than all other forms of the plant. Animals in group B1, B2, B3, C1 and C2 showed anthelmintic efficacy of 19.47%, 23.58%, 31.66%, 31.76% and 33.33% from day 0 to day 15th post-treatment. Gastrointenstinal nematodes of sheep have produced anthelmintic resistance against Levamisole at the dose rate of 7.5mg/kg. In previous studies Levamisole had showed efficacy of 99.99%, 99% and 98%. It is therefore recommended that further investigation on huge scale should be passed out concerning a great number of animals, quantities higher than those used in the present study, documentation of active principles, and calibration of dose and toxicity studies for drug development from the herbal plant. Availability: Items available for loan: UVAS Library [Call number: 2688-T] (1).

31. Comparison Of The Cryoprotective Effect Ofquailand Chicken Egg Yolk On The Freezability Of Nili-Ravi Buffalo Bull Spermatozoa

by Kamran Khaliq (2014-VA-812) | Prof. Dr. Nasim Ahmad | Prof. Dr. Mian Abdul Sattar | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Pakistan is one of the most important agricultural country. Livestock as a subsector of agriculture contributes 11.4% in the national economy and 56.3% in agriculture. Buffalo is honored as the black gold of Pakistan and Nili-Ravi is the most important animal in this regard. Approximately 60% of the total milk produced within a country comes from buffalo. Artificial insemination is considered to be the most important tool for the prompt genetic improvement of livestock. Mammalian spermatozoa undergo many structural and biochemical changes during cryopreservation which may leads to an impaired fertility. Buffalo bull spermatozoa are sensitive and more prone to damage than cattle. Various experiments have been conducted in order to improve the post thaw semen quality which includes supplementation of egg yolk from different bird species that tends to work as a cryoprotectent against cold shock. In the present study it is assumed that replacement of chicken egg yolk with quail egg yolk in cryodiluents improves the freezability of Nili-Ravi buffalo bull spermatozoa. Three adult Nili-Ravi buffalo bulls without any clinical reproductive anomaly and kept at Semen Production Unit (SPU), Qadirabad District Sahiwal were used in this study. Semen from these three experimental bulls was collected twice a day twice a week by using an artificial vagina already maintained at 42 ºC. Each bull was subjected to a minimum of seven replicas. Both semen samples were collected on each collection day from each of the three experimental bulls with an interval of about ten minutes between the ejaculates. After initial evaluation for sperm motility and concentration, both ejaculates collected from the same bull at the same collection day were pooled and again evaluated for sperm cell concentration. Every pooled semen sample was then divided into five aliquots and extended with one of the five experimental extenders namely A, B, C, D, and E in order to maintain a final concentration of 40 million spermatozoa per 0.54 ml of diluted semen. Instantly after dilution motility percentile for each semen sample was recorded. After filling, open ends of straws were sealed with polyvinyl pyrolidine powder and allowed to cool and then it was stored at 4-5 ºC for equilibration. Finally the samples were frozen and stored in liquid nitrogen. On post thaw evaluation, motility, live/dead count, plasma membrane integrity, acrosomal integrity, and DNA integrity along with CASA evaluation parameters for motility characteristics were performed. Data were analyzed by means of two way ANOVAand comparison between means was done by using Duncan’s multiple range test. Results showed that post-extension motility and DNA integrity did not differ significantly between extender A and E. While post-thaw motility, plasmolemma intactness, acrosomal integrity, and viable sperm count was found to be higher in extender E compared to extender A. Similarly, CASA evaluation factors like progressive sperm cells motility, straight line velocity, straightness, linearity index, and the percentile of rapidly moving spermatozoa were also significantly different among extender E and A. While curvilinear velocity, and amplitude of lateral head displacement was higher in extender B that contained 5% QEY. So it was concluded that substitution of 20% CEY with 20% QEY in cryodiluents resulted in an improved post-thaw motility, viable sperm count, plasmolemma intactness, and sperm kinematics. Furthermore, it was seen that reduction in the concentration of QEY from 20% to 5% resulted in decreased sperm motility characteristics. Availability: Items available for loan: UVAS Library [Call number: 2686-T] (1).

32. Comparison Of Cold Versus Warm Blood Cardioplegia In Diabetic Patients Undergoing Coronary Artery Bypass Grafting On Cardiopulmonary Bypass

by Komal Saeed Awan (2014-VA-953) | Prof. Dr. Habib ur Rehman | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: In cardiopulmonary bypass, inflammatory reaction started in the body as the blood cells comes in contact with the non­physiological surfaces. This response is more prominent in high risk diabetic patients. Since warm cardioplegia is more prone to an incidence of stroke, increase level of cardiac enzymes and is more controversial. However, cold blood cardioplegia gives better myocardial protection, less inotropic support and inflammatory response. Data were collected on pre designed proforma for the pre operative, per operative and post operative findings. Consecutive 140 patients were included in this study after fulfilling the inclusive criteria. In this seven day trial we compared hospital outcomes of diabetic patients undergoing CABG (coronary artery bypass grafting) on cardiopulmonary bypass with either intermittent antegradge cold or warm blood cardioplegia. Outcomes of this trial were divided into primary and secondary end points. Hospital mortality was primary end point and need of ionotropic support, left ventricular function, infection, inflammation and cardiac enzymes were secondary end point. Ionotropic support data indicated that cold blood cardioplegia is better than warm blood cardioplegia. Results have demonstrated that risk factors like smoking, hyperlipidemia, renal failure has shown no significant results in both cardioplegia techniques. Ventricular assist device Intra aortic balloon pump has shown no significant results. Use of inotropes has shown significant results in both groups like adrenaline, nor-adrenaline and dopamine doses were higher in WBC. Post operative ejection fraction has shown no significant results between groups. Effect of cardiac enzymes and c-reactive protein has shown significant results in both groups. Results have demonstrated that in CBC Cardiac enzymes and inflammatory response were significantly less. Post operative blood parameters hemoglobin, serum creatinine and blood sugar level have shown significant differences between groups. Bypass time and aortic cross clamp have shown no effect between groups. The result of this study indicated that cold blood cardioplegia is superior over warm blood cardioplegia in cardiopulmonary bypass diabetic patients. Availability: Items available for loan: UVAS Library [Call number: 2747-T] (1).

33. Comparison Of Cold Versus Warm Blood Cardioplegia In Diabetic Patients Undergoing Coronary Artery Bypass Grafting On Cardiopulmonary Bypass

by Komal Saeed Awan (2014-VA-953) | Prof. Dr. Habib ur Rehman | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: In cardiopulmonary bypass, inflammatory reaction started in the body as the blood cells comes in contact with the non­physiological surfaces. This response is more prominent in high risk diabetic patients. Since warm cardioplegia is more prone to an incidence of stroke, increase level of cardiac enzymes and is more controversial. However, cold blood cardioplegia gives better myocardial protection, less inotropic support and inflammatory response. Data were collected on pre designed proforma for the pre operative, per operative and post operative findings. Consecutive 140 patients were included in this study after fulfilling the inclusive criteria. In this seven day trial we compared hospital outcomes of diabetic patients undergoing CABG (coronary artery bypass grafting) on cardiopulmonary bypass with either intermittent antegradge cold or warm blood cardioplegia. Outcomes of this trial were divided into primary and secondary end points. Hospital mortality was primary end point and need of ionotropic support, left ventricular function, infection, inflammation and cardiac enzymes were secondary end point. Ionotropic support data indicated that cold blood cardioplegia is better than warm blood cardioplegia. Results have demonstrated that risk factors like smoking, hyperlipidemia, renal failure has shown no significant results in both cardioplegia techniques. Ventricular assist device Intra aortic balloon pump has shown no significant results. Use of inotropes has shown significant results in both groups like adrenaline, nor-adrenaline and dopamine doses were higher in WBC. Post operative ejection fraction has shown no significant results between groups. Effect of cardiac enzymes and c-reactive protein has shown significant results in both groups. Results have demonstrated that in CBC Cardiac enzymes and inflammatory response were significantly less. Post operative blood parameters hemoglobin, serum creatinine and blood sugar level have shown significant differences between groups. Bypass time and aortic cross clamp have shown no effect between groups. The result of this study indicated that cold blood cardioplegia is superior over warm blood cardioplegia in cardiopulmonary bypass diabetic patients. Availability: No items available

34. Molecular Diagnosis Of Brucella Zoonosis As Blood Transfusion Hazard In Metropolitan Population Of Lahore And Okara

by Amna Azam (2011-Va-3560 | Dr. Wasim Shehzad | Dr. Iahtasham Khan | Dr. Muhammad Imran | Dr. Imran Rashid.

Material type: book Book Publisher: 2017Dissertation note: Brucellosis and Coxiellosis are one of the most spreading zoonotic diseases. They both are facultative, intracellular, Gram negative and involved in bioterrorism and agro-terrorism attacks. Brucella abortus and Brucella melitensisare common among all specie of Brucella. Total two hundred Human Blood transfusion samples were collected from hospitals of Okara and Lahore. Blood was collected in vacutainers (without EDTA). After serum isolation serological test RBPT were performed of all samples. Eighty nine out of two hundred were positive to RBPT with seroprevalence of 44.5% (95% Confidence Interval [CI]: 37.61 – 51.4). DNA extraction was done. The concentration of DNA was analyzed through Nanodrop 2000. Then these samples were subjected to Genus specific Real-time PCR analysis. Forty one out of two hundred were positive to Genus specific Real-time PCR with seroprevalence of 20.5% (95% Confidence Interval [CI]: 14.9 – 26.09). Brucella genus positive samples were subjected to two species specific PCR Brucella abortus and Brucella melitensis respectively. Forty one out of forty one Brucella genuspositive samples were positive to Brucella melitensisand none of the sample was positive to Brucella abortus. These two hundred DNA samples were then subjected to Coxiella specific Real-time PCR analysis and 4 were found positive with seroprevalence of 2% (95% Confidence Interval [CI]: 0.06 – 3.94). Results were recorded in the form of Ctvalue. Results indicate that Real-time PCR is more efficient than RBPT and due to increasing seroprevalence in Blood transfusion samples its screening should be included in normal blood transfusion screening procedure through collaboration with Government to prevent transfusion transmitted infections (TTI’s). Availability: Items available for loan: UVAS Library [Call number: 2873-T] (1).

35. In-Vitro Effect Of Chromium Chloride On The Electrophysiological Indices Of Jejunum In Broilers.

by Maria Khalid Majeed (2014-VA-961) | Prof. Dr. Habib ur Rehman | Dr. Sajid Khan Tahir | Dr. Wasim Shehzad.

Material type: book Book Publisher: 2017Dissertation note: Chromium is a well-known component of glucose tolerance factor, which participates in glucose metabolism by enhancing the effects of insulin. The Ussing chamber is a very powerful technique to study ion transport across tissues. This study is to determine the effect of chromium chloride hexahydrate on electrophysiology of jejunum and its interaction with glucose transport. According the background we concluded the hypothesis i-e Chromium chloride may enhance the glucose transport in jejunum mucosa of the broilers. To approach this hypothesis, we performed an experiment into different groups. In overall experiment, we take Twelve broilers of 35 days of age was procured from local farm. After a week, birds were slaughtered; jejunum section of the birds were isolated then stripped off serosal layer. Each segment of the intestine was mounted and labelled on ussing chamber separately with groups viz, Group1 (0μM, control), Group2 (5μM CrCl3), Group3 (10μM CrCl3). After baseline, add CrCl3 to the mucosal side of UC and note the reaction. Then Add glucose after incubation period. The peak response obtained within 2-3 mins. Then the change in Isc (ΔIsc) was calculated approximately after addition of CrCl3 and glucose. The PD, Isc, and Gt were measured by voltage/current clamp. The data was analysed using one-way ANOVA and was presented as mean±SEM. Group differences were compared by Duncan multiple range Test (P<0.05). The current study showed that there are no significant differences shown in initial short circuit current and ▲Isc and tissue conductance before and after adding the chemicals. In conclusion, it is hoped that results presented in the current research would open avenues for further discussion and more concerted investigations into each of the area covered. More specifically, there are no previous research reports on the in vitro effect of chromium chloride hexahydrate in poultry. Availability: Items available for loan: UVAS Library [Call number: 2869-T] (1).

36. Identification Of Pkhd1 Gene Mutation In Polycystic Kidney Disease And In-Silico Molecular Characterization In Different Mammals

by Taslim Un Nisa (2015-VA-1105) | Dr. Wasim Shehzad | Dr Muhammad Yasir Zahoor | Prof.Dr. Aftab Ahmad Anjum.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Fibrocystin is a large, receptor-like protein that is involved in the tubulogenisis and maintenance of duct-lumen architecture of epithelium. Fibrocystin has a combination with the primary cilia of epithelial cell. Renal tubules (small tube) of kidney where urine is formed lined by tiny hair like projection. Twenty five suspected patient was selected and DNA extracted through organic extraction method from the suspected patient blood. Primers were designed PKHD1gene’s coding sequence located at 6p12.2 in human. The coding region sequenced using the ready mate terminator Sequencing Reaction Kit by Perkin Elmer/ABI and read in an automated sequencer. The allele’s variants have been only reported for Fibrocystin protein in human. All of the sequences are evaluated by using Chromas and Bioedit software for sequence analysis. The in-silico protein analysis is done for normal and mutated alleles through UCSC and RAPTROX. Homology analysis was also done between human and mammals DNA sequence. We found mutations which are associated with ARPKD disease and these variants are most common in other population whereas we also found some new variants. There are some reported mutations which we found in our study such as (c3790C>T),(c3891G>T),(c3790C>T). We found three new mutations in PKHD1 gene. The new mutations which we found are (c3681G>A),(c3804C>T),(c3931A>C).These mutations (c3790C>T) and (c3931A>C) in the exon 32 show significant effect on the gene and protein function. Geneticanalysis of PKHD1gene show thatPakistanifamilies have mutations as compared to other population along with some common exonic regions such as exon 32 whichisalsodescribed by others in two different studies.We also analyze the pedigrees of these patients which are consanguineous families and autosomal recessive polycystic disease. We found total six mutations in this gene including missense/ synonymous mutations. In which, three novel mutations and others are reported mutations. These variations from the results are due to the population and consanguineous families’ pattern.In our study, we also found that Mouse and Chickencan preferably be used as a modal organisms in pathology.This study will also help us in the development of molecular genetic testing for their detection in Pakistani families and population. Availability: Items available for loan: UVAS Library [Call number: 2941-T] (1).



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