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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).

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2.
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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).

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3.
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Genetic Study Of Slc6a4 Gene In Convicted Offenders From Prisons Of Punjab, Pakistan Exhibiting Antisocial Personality Disorder Traits

by Asima Saman | Dr. Saadat Ali | Dr. M. Yasir Zahoor | Dr. M. Imran.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Antisocial Personality Disorder (ASPD) is characterized by incapacity of an individual to adapt themselves to social norms. Patients with ASPD typically have irritability problems and aggressive feelings toward other people. The serotonin transporter gene (5-HTT orSLC6A4) has been associated with regulation of serotonergic neurotransmission, mood and behaviour traits.Low levels of the neurotransmitter serotonin are believed to affect judgement, planning, and impulse control in ASPD sufferers. Genetic polymorphism in selected intronic region of serotonin transporter SLC6A4 gene was associated with antisocial personality disorder (ASPD) in convicted criminal of Punjab, Pakistan. We have selected two regions from SLC6A4 gene, which was intron 1 and exon 3.After extraction of DNA Polymerase Chain Reaction was used to amplify the extracted DNA of criminals (n=20) and control (n=10) for selected segments of intron 1 and exon 3 of SLC6A4gene. Sanger’s DNA sequencing method (di-deoxy chain termination method) was used to sequence the amplified fragments. Statistical and bioinformatics tools were used to analyse the data. Intron 1 has shown 5-HTTLPR polymorphism S allele (0.85 frequencies), LA allele (0.05 frequencies) and LG allele (0.1 frequencies) and exon 3 did not show polymorphism in criminal’s sample. The study highlights the role of SLC6A4 gene polymorphism in criminals of Punjab having antisocial personality disordertraits. Availability: Items available for loan: UVAS Library [Call number: 2576-T] (1).

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4.
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Development Of Novel Mtdna Metabarcodes For The Species Differentiation Of Class Aves

by Syeda Rida Mehak Sherazi (2010-VA-477) | Dr. Muhammad Imran | Dr. M. Yasir Zahoor | Mr. Shahid Abbas.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: The Folmer COI mtDNA universal primers that are considered standard for DNA barcoding of life contain so many mismatches against the target sequences of vertebrate origin that they often end in failure to amplify many of vertebrate DNA extractions. This discrepancy favors for the selection and designing of new metabarcode primers that can be used to identify all individuals of vertebrates or at least all individuals represented in a class of Vertebrata such as Class Aves. The current study embarks on such an endeavor. In this study development of new mtDNA metabarcode (16SrRNA) that can be used as universal primers to amplify almost all species of Class Aves for different forensic and molecular biodiversity analyses. Blood/feather/tissue samples were collected from Class Aves (one specimen from every order reported to be present in Pakistan). DNA was extracted from the collected specimens through standard organic method, qualified and quantified and then PCR-amplified using novel universal primers selected from aligned mtDNA sequences originating from all Aves mitochondrial DNA genomes submitted to different online sequence databases such as NCBI nucleotide database. The sensitivity of PCR also be assessed using a range of DNA concentrations. The amplified products were sequenced on ABI Genetic Analyzer following Sanger’s dideoxy method of sequencing. The correctness of obtained mtDNA sequences were examined visually in Chromas Lite 2.1 software and then alignment of these sequences were performed against highly similar DNA sequences in NCBI nucleotide databases using BLAST in order to identify origin of unknown mtDNA sequences. With the help of sequencing and phylogenetic studies specificity of the universal primer set confirmed and presented as a novel metabarcode (16SrRNA) for species level identification of large number of Avian species In summary, we present universal method for species classification of Aves using a targeted parallel sequencing approach. Both sequencing and phylogenetic studies experiments confirm Summary 82 specificity of universal primer set. Although promising results were obtained with current settings, rapid improvement of bench top instruments will further develop method with less hands-on, fewer sequencing errors and lower detection limit. So, in future, this barcode can be used for species identification in various fields of study such as meat adulteration, illegal trade, food mislabeling and molecular estimation of biodiversity. Availability: Items available for loan: UVAS Library [Call number: 2617-T] (1).

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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).

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