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1. Comparative Evaluation Of Anti-Hyperglycemic Effect Of Herbal Medicinal Plants Extracts On Alloxan Induced Diabetic Albino Rats

by Sonia Aziz (2011-VA-03) | Ms. Asma Waris | Ms. Faiza Masood | Dr. Maryam Javed.

Material type: book Book; Literary form: not fiction Publisher: 2014Dissertation note: Diabetes mellitus is a clinical syndrome described as inappropriate hyperglycemia triggered by a relative or absolute deficiency of insulin or by a resistance to the action of insulin at the cellular level. It is the most shared endocrine disorder, upsetting 16 million individuals in the United States and as many as 200 million worldwide (Debra, 1991). The word diabetes was devised by the Greek physician Aeretaeus in the first century A.D. In the 17th century, Willis detected that the urine of diabetics as ideally sweet as if infused with honey or sugar. The existence of sugar in the urine of diabetics was established by Dobson in 1755 (Straton et al. 2000). Diabetes mellitus is a global health crisis, which has been obstinately disturbing the humanity, regardless of the socioeconomic profile and geographic location of the population. According to an estimate, one person is identified with diabetes every 5s somewhere in the world, while someone dies of it every 10s. Diabetes mellitus has achieved a pandemic form. Hence, it is very vital to control diabetes and its difficulties to lessen the human suffering (Wild et al. 2004). Alloxan a glucose equivalent and is toxic by selectively abolishing insulin-producing cells in the pancreas (that is beta cells) of many animal species. This produces an insulin-dependent diabetes mellitus (called "Alloxan Diabetes") in these animals, with features similar to type 1 diabetes in humans. Alloxan is selectively toxic to insulin-producing pancreatic beta cells because it preferentially amasses in beta cells through uptake via the GLUT2 glucose transporter. Alloxan, in the presence of intracellular thiols, produces reactive oxygen species which start toxicity by its redox reaction (Lenzen et al. 1998). There are diverse methods to the management of diabetes, like insulin treatment in type 1 diabetes: Sulphonylureas, which discharge insulin from pancreas by blocking the ATP-sensitive potassium channels; Biguanides, which reduce the insulin resistance; Thizaolidinediones, which upsurge the insulin sensitivity; alpha-glucodase inhibitors like acarbose, which lessen glucose absorption from intestine, thus reducing postprandial hyperglycemia; metiglinides like repaglimide and nateglamide, which are insulin secretogogues (Aslam, 1988). In spite of the statistic that synthetic drugs such as insulin, investigators have been building efforts to find insulin-like substances from plant sources for the treatment of diabetes. More than 1200 plant species have been suggested for the managementof diabetes (Radha et al. 2011) Natural resources such for example plants are cherished source of bioactive compounds. A large number of drugs have been recognized in medicinal practice from natural products (Philipeon et al. 2010). Recent scientific research and clinical studies have established the usefulness of some medicinal plants and herbal preparations in the development of standard glucose homeostasis. Herbal treatment have been used in patients with insulin-dependent and non-insulin dependent diabetes, diabetic retinopathy, diabetic peripheral neuropathy and other penalties of this metabolic disease (Ahmed et al. 2006). The herbal drugs are recommended extensively because of their effectiveness, less side effects and comparatively low cost (Lezney et al. 2004). Ethno pharmacological reviews show that more than 1200 plants are used in customary medical systems for their suspected hypoglycemic activity (Marles and Farnsworth, 1995, Dey et al. 2002, Grover et al. 2002). The hypoglycemic activity of a huge number of these plants/plant products has been appraised and inveterated in animal models (Gupta et al. 2005, Kesari et al. 2006) as well as in human beings (Herrera et al. 2004, Jayawardena et al. 2005). In some circumstances the bioactive principles have also been secluded and identified. However, the mechanism of action whereby most of these plants and yields lesser the blood glucose level rests hypothetical. This study reveals the comparative effect of different herbal plants effect on alloxan induced diabetic rats. Six different herbal plants have been used in this study to investigate the hypoglycemic activity. These plants areAllium sativum (Garlic), Aloe vera(Kanwargandal), Gymnemasylvestre (Gurmar), Momordicacharantia (karela), Trigonellafoenum-graecum (Methidana), and Syzigiumcumini(Jamun). Table 1: Plants used in present study Plant and family Plant part used Active ingredient Mechanism of action Reference Allium sativum, Alliaceae Garlic gloves S-methyl cysteine sulphoxide-precursor of allicin and garlic oil Arouse in vitro insulin discharge,Hinder glucose making by the liver Sheela et al. 1992, Augusti and Shella 1996. Aloe vera, Aspholedeceae Leaf pulp Phytosterols Excite production or discharge of insulin Modify action of carbohydrate processing enzymes Rajasekaran et al. 2004, Tanaka et al. 2006. GymnemaSylvestre, Asclepiadaceae Leaves Gymnemosides and gymnemic acid (from the saponin fraction) Triterpene glycosides Kindle exudation of insulin from rat islets. Declines the activity of gluconeogenicenzymes,Induce beta cell regeneration. Shanmugasundaram, 1990, Chattopadhyay, 1999. Mormordicacharrantia, Cucurbitaceae Fruit pulp Charantin (a peptide),Insulin like polypeptide P ("vegetable insulin") Encourage insulin secretion, Quash the activities of gluconeogenic enzymes Rises the quantity of beta cells in diabetic rats Rao et al. 1981, Day et al. 1992, Sarkar et al. 1996. Trigonellafoenumgraecum, Fabaceae Seeds  Alkaloid-trigonelline, nicotinic acid, and coumarin  4hydroxyisoleucine Galactomannan depress digestion and absorption of carbohydrates Upsurge glucose induced insulin release Khosla et al. 1995, Hannan et al. 2007. Syzigiumcumini Seeds  Mycaminose Kindle kinases intricate in peripheral utilization of glucose  Achrekar et al. 1992.  Kumar et al. 2008 1.1: Allium sativum(Garlic) is a common zesty flavoring agent used since prehistoric times. Garlic has been cultured in all over world for its distinctive flavor, foodstuff, and medicinal properties. It has mostly been ascribed to its hypoglycemic, anticoagulant, antibiotic, hypo-cholesterolaemic, antihepatotoxic, anticancer, immune system modulatory and antioxidant possessions (Bakri and Douglas, 2005). Figure 1: structure of allicin 1.2: Aloe vera(Kanwargandal)is one of the therapeutic plants which are conventionally well accredited plant in the controlling of diabetes. It fits to family Liliaceae (sub-family of the Asphodelaceae). Many studies titles that the high innards of phenolic compounds, glycosides (aloins), 1,8-dihydroxyanthraquinone derivatives,β -1,4 acetylated mannan, mannose-phosphate and alprogenglucoprotein in the A. vera is vital for its biotic action. Through past two years, Aloe vera used as helpful beneficial agent which defensively act as a free radical scavenging and other antioxidant characteristics on diabetic patients, by monitoring raised anions in an alloxan or STZ-induced diabetic animal models (Nakamura, et al. 2011). Figure 2: structure of phytosterole 1.3: Gymnemasylvestre (Gurmar) is a plant used in Asia as a usual cure for diabetes or “sweet urine.” The hypoglycemic action of Gymnema leaves was first recognized in the late 1920s. Gymnema is testified to upsurge glucose uptake and utilization. It also mends the utility of pancreatic β-cells and may also decline glucose captivation in the gastrointestinal tract. Phytochemically the plant has been described to comprehend gymnemagenin- the sapogenin. Gymnemic acid was sequestered in pure states from the hot water extract of leaves of G. sylvestre (Puratchimani and Jha, 2004). Figure 1: structure of gymnemic acid 1.4: Momordicacharantia (Bitter Melon) also known as karela, is one of the plants normally used for its glucose-lowering properties (Ahmed et al., 1998). The slices of the plant usually used contain the entire plant, its fruit or seeds, all of which are bitter due to the manifestation of the chemical momordicin. The anti-diabetic constituents in bitter melon comprise charantin, vicine, polypeptide-p, alkaloids and other non-specific bioactive components such as anti-oxidants (Beloin, et al. 2005). Figure 4: structure of momordicin 1.5: Trigonellafoenum-graecum (Fenugreek L. Leguminosae) is one of the ancient therapeutic plants, originating in India and Northern Africa.The leaves and seeds, which ripe in long pods, are used to formulate extracts or powders for medicinal use.The hypoglycemic properties of fenugreek have been recognized to numerous mechanisms.The amino acid 4-hydroxyisoleucine in fenugreek seeds amplified glucose-induced insulin release in human and rat pancreatic islet cells. Fenugreek seeds apply hypoglycemic effects by exciting glucose-dependent insulin discharge from pancreatic beta cells, as well as by impeding the actions of alpha-amylase and sucrase, two duodenal enzymes involved in carbohydrate breakdown (Gupta, et al. 2001). Figure 5: structure of 4-hydroxyisolucine 1.6: Syzigiumcumini(Jamun) tree belongs to the Myrtaceae family. This is also called as Jamun, Jambul and Jambol in Pakistan, India and Malaya. The barks, leaves and seeds extracts of SC have been testified to have anti-hyperglycemic, anti-inflammatory, antibacterial and anti-diarraheal effects. A complex mycaminose is extracted from its seeds which display anti-diabetic characteristic (WL Li, et al. 2004). Figure 6: structure of mycaminose There are numerous potential mechanisms through which these herbs can perform to regulate the blood glucose level (Tanira, 1994). The mechanisms of action can be associated, commonly to the capability of the plant in question (or its active principle) to lesser plasma glucose level by meddling with one or more of the procedures involved in glucose homeostasis. The described mechanisms whereby herbal antidiabetic remedies decrease blood glucose levels are more or less alike to those of the artificial oral hypoglycemic drugs and are abridged as follows (Acharya et al. 2008, Bastaki, 2005, Bnouham et al. 2006). i) Stimulation of insulin production and/or discharging from pancreatic beta-cells ii) Revival of impaired pancreatic beta cells iii) Development of insulin sensitivity iv) Imitating the action of insulin v) Modification of the action of some enzymes that are tangled in glucose metabolism reducing the absorption of carbohydrates from the gut. The effectiveness of herbal drugs is substantial and they have insignificant side effects than the synthetic antidiabetic drugs. There is growing demand by patients to use the natural products with antidiabetic activity. In recent times there has been improved concern in the plant remedies. Plants grasp certain potentials in the organization of Diabetes mellitus. Isolation and documentation of active ingredients from these plants, preparation of unvarying dose and dosage schedule can play a noteworthy role in improving the hypoglycemic action (Jung et al. 2006). Availability: Items available for loan: UVAS Library [Call number: 2209,T] (1).

2. Identification Of Genetic Variations In Toll Like Receptor 1(Tlr-1) Gene To Evaluate Its Potential For Enhanced Resistance To Bovine Tuberculosis

by Shehar Bano (2013-VA-09) | Dr. Maryam Javed | Prof. Dr. Tahir Yaqub | Miss Huma Mujahid.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2015Dissertation note: Bovine tuberculosis is a disease caused by the species included in the Mycobacterium tuberculosis complex. Toll-like receptors (TLRs) are a family of conserved innate immune recognition receptors that trigger adaptive immune responses. TLR1 play an important role in host defense against mycobacteria, especially by mediating the response to mycobacterial triacylated lipopeptides. The objective of this study is the identification of single nucleotide polymorphisms within the coding region of TLR1 gene to evaluate its potential for enhanced the resistance to bovine tuberculosis in Nili-Ravi buffalo breed. Fifty blood samples of Nili-Ravi breed were collected from UVAS Pattoki Campus, Research Farm B and Buffalo Research Institute (BRI) Pattoki. Inorganic method was used for DNA extraction, for amplification of the coding region of TLR1 gene PCR (Polymerase Chain Reaction) was used using specially designed primers and the PCR products were sequenced through Sanger’s Chain Termination method. For the analysis and alignment of sequencing the results obtained after sequencing were analyzed and aligned using the CLUSTAL W and BLAST software. After all these analysis Ten SNPs were identified in the coding region of TLR1 mentioned in table. The Ten SNPs identified in the coding region of TOLL LIKE RECEPTOR 1 were in this order P1 C>T, P2 T>C, P3 T>C, P4 T>C, P5 T>C, P6 C >T, P7 T>C, P8 C >T, P9 A>G and P10 A>G. The one SNP found in the current research is in compliance with the (Sun et al. 2012) research on TOLL LIKE RECEPTOR 1 hence Nine SNPs found in the current research are novel in Nili Ravi buffalo. The SNPs in the exonic region that is P1 C>T, P2 T>C, P3 T>C, P4 T>C, P5 T>C, P6 C >T, P7 T>C, P8 C >T, P9 A>G and P10 A>G were all transitions i.e. the conversion of purines to purines. Population genetic analysis and allelic distribution at all loci was analyzed using POPGENE 32 software indicated that at [P3=0.243009> 0.05] followed the assumptions of the Hardy-Weinberg equilibrium indicating that the alleles were randomly distributed throughout the population, no migration had occurred, no bottlenecks happened and population remained large in numbers.This Non-significant and obeying HWE, so can be potential marker for genetic selection.At [P1= 0.040418< 0.05], [P2=0.033603< 0.05], [P4=0.000649< 0.05], [P5=0.000262< 0.05], [P7=0.015112< 0.05] and [P9=0.000111< 0.05] the probability value below 0.05 indicated that population at these polymorphic sites was not obeying Hardy-Weinberg equilibrium. This indicated that at these positions alleles were not equally distributed in population. It can be concluded from my research that the SNPs identified in the current research may also hold potential for marker-assisted breeding programs, with the aim of breeding more BTB-resistant animals and herds within both the national farms and the private sector. Availability: Items available for loan: UVAS Library [Call number: 2335-T] (1).

3. Mutation Anlysis Of DTNBP1 Gene In Pakistani Patients With Schizophrenia Disorder

by Hafiza Sidrah Yasin (2013-VA-11) | Dr. Asif Nadeem | Dr. Maryam Javed | Dr. Abu Saeed Hashmi.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Schizophrenia (SCZ) disorder is a mental complex, heterogeneous and chronic neurodegenerative disorder with a cumulative prevalence of 1%. SCZ is an idiopathic disorder of the cortex and hippocampus. Environmental as well as genetic factors contribute to its complex pathogenesis. A functional repeat polymorphism in the Dystrobrevin Binding Protein 1 (DTNBP1) gene promoter conveys susceptibility for SCZ disorder. The DTNBP1 has been implicated in rare autosomal dominant forms of SCZ disorder because of mutations associated with severe disease progression and a typical physical signs and symptoms, indicative of neurodegeneration. Mutation in DTNBP1 gene has association with change in dysbindin protein which leads to change in abnormal neurotransmitter trafficking which leads to decrease in neuronal size, brain atrophy and reduced glutamate release in schizophrenia disorder. A systematic approach was applied to proceed the present study in order to identify the single nucleotides polymorphisms in schizophrenic patients. Blood samples (n=40) were collected from schizophrenia disorder patients. DNA was extracted by organic method. Primers were designed using Primer3 software. The amplification of gene was done by Polymerase Chain Reaction. PCR products were sequenced bi-directionally on ABI 3130XL Genetic analyzer. Sequence alignment was performed for polymorphism identification. The analysis of identified polymorphism was done by CHROMAS software. Sequence was aligned by Blast tool of NCBI. Difference between allele and genotype frequency of studied gene was evaluated and analyzed by using “SNPator”. The present study provides information about the susceptibility and genetic basis of the individual towards this disease and identified polymorphisms provides the opportunity to diagnose the disease earlier on the basis of particular SNPs in Pakistani patients. Availability: Items available for loan: UVAS Library [Call number: 2382-T] (1).

4. Genetic Effect Of Interferon Gamma On Bovine Resistance Against Mycobecterium Bovis

by Syed Ahmed Raza Rizvi (2012-VA-819) | Dr. Maryam Javed | Dr. Tanveer Hussain | Dr. Muhammad Tayyab.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Bovine tuberculosis is a disease caused by the species included in the Mycobacterium tuberculosis complex. IFN-GAMMA are a family of conserved innate immune recognition receptors that trigger adaptive immune responses. IFN-GAMMA play an important role in host defense against mycobacteria, especially by mediating the response to mycobacterial triacylated lipopeptides. The objective of this study is the identification of single nucleotide polymorphisms within the coding region of IFN-GAMMA gene to evaluate its potential for enhanced the resistance to bovine tuberculosis in Nili-Ravi buffalo breed. Fifty blood samples of Nili-Ravi breed were collected from UVAS Pattoki Campus, Research Farm B and Buffalo Research Institute (BRI) Pattoki. Inorganic method was used for DNA extraction, for amplification of the coding region of IFN-GAMMA gene PCR (Polymerase Chain Reaction) was used using specially designed primers and the PCR products were sequenced through Sanger’s Chain Termination method. For the analysis and alignment of sequencing the results obtained after sequencing were analyzed and aligned using the CLUSTAL W and BLAST software. After all these analysis Ten SNPs were identified in the coding region of IFNG mentioned in table. The Eight SNPs identified in the coding region of INTERFERON GAMMA were in this order P1 C>T, P2 T>C, P3 T>C, P4 T>C, P5 T>C, P6 C >T, P7 T>C, and P8 C >T. The one SNP found in the current research is in compliance with the (Sun et al. 2012) research on INTERFERON GAMMA hence Nine SNPs found in the current research are novel in Nili Ravi buffalo. Research on IFN-GAMMA hence Seven SNPs found in the current research are novel in Nili Ravi buffalo. The SNPs in the exonic region that is P1 C>T, P2 T>C, P3 T>C, P4 T>C, P5 T>C, P6 C >T, P7 T>C, P8 C >T, among them four were transitions and four were transversion . Population genetic analysis and allelic distribution at all loci was analyzed using Summary 57 POPGENE 32 software indicated that at [P3=0.354539>0.05] , [P5=0.365524>0.05]followed the assumptions of the Hardy-Weinberg equilibrium indicating that the alleles were randomly distributed throughout the population, no migration had occurred, no bottlenecks happened and population remained large in numbers. This Non-significant and obeying HWE, so can be potential marker for genetic selection. At [P1= 0.000032< 0.05], [P2=0.038766< 0.05] and [P7=000394< 0.05] the probability value below 0.05 indicated that population at these polymorphic sites was not obeying Hardy-Weinberg equilibrium. This indicated that at these positions alleles were not equally distributed in population. It can be concluded from my research that the SNPs identified in the current research may also hold potential for marker-assisted breeding programs, with the aim of breeding more BTB-resistant animals and herds within both the national farms and the private sector. Availability: Items available for loan: UVAS Library [Call number: 2419-T] (1).

5. Characterization And Phylogenetic Analysis Of Hemagglutinin Gene Of Avian Influenza Virus Subtype H9n2 Isolated In 2015

by Arslan Mehboob (2009-VA-76) | Prof. Dr. Tahir Yaqub | Dr. Jawad Nazir | Dr. Maryam Javed.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: H9N2 Avian influenza outbreaks has caused great economic losses to poultry industry resulting in decrease egg production, high morbidity and mortality. Due to different antigenic variants H9 has become problematic. It has the ability to cross species barrier and increase in pathogenicity. Hemagglutination inhibition (HI) test is employed extensively for subtyping and detection of antibody titre against the virus. Continuous mutations in the HA gene transforms AIV subtype H9N2 into more pathogenic virus that may have pandemic potential and can cross species barrier. Thus, it was necessary to identify various antigenic variants of H9 virus. It was important to study the HA gene as it plays vital role in viral attachment, release of genetic material and pathogenicity. In present study, a sum of four H9 virus samples were isolated. Both serological and molecular confirmation was done. 200 samples from different areas were collected and properly labelled. They were then processed for egg inoculation in embryonated eggs. Virus was grown in embryonated eggs and harvested fluid is then proceeded for confirmatory testing. Haemagllutination and Haemagllutination Inhibition testing was done. RNA was extracted by Kit method and cDNA was synthesized. Reverse Transcriptase (RT-PCR) was performed using specific primer sets and then the amplicon were run on agarose gel. The bands obtained was sent for sequencing and Phylogenetic analysis was obtained using software and tree was constructed. Protein analysis was also performed. The present study enabled us to characterize and construct Phylogenetic tree of HA gene of currently prevailing H9N2 Avian Influenza isolates in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2474-T] (1).

6. 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).

7. Seroprevalence And Risk Factor Analysis Of Bluetongue Virus In Lahore And Faisalabad Districts Of Punjab Province, Pakistan

by Syeda Marriam Maqbool (2014-VA-522) | Dr. Muhammad Zubair Shabbir | Dr. Ali Ahmad Sheikh | Dr. Maryam Javed.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Domestic animals play an important role in the rural and agricultural economies of developing countries. Therefore, animal diseases pose a threat to country’s economy, animal welfare, the environmental and public health. One of the important animal diseases is Bluetongue, listed as notifiable disease by OIE. Causative agent is Bluetongue virus (BTV) an arbovirus that belongs to genus Orbivirus with in family Reoviridae. The main route of transmission is through the bite of Culicoides biting midges. Disease is enzootic and widely distributed in areas where susceptible animals and vector species are prevalent. It has been reported worldwide including the neighboring countries of Pakistan. BTV is also considered an endemic in Pakistan but little information is available on its epidemiology in this area. Serological tests can detect antibodies produced against infection and helpful to analyze the prevalence of a pathogen in circumstances when there lacks vaccination practice to ruminants in a given geographical area. Competitive ELISA was used to identify antibodies to BTV in the sera samples of animals in Faisalabad and Lahore districts. Blood samples were collected from randomly selected villages of both districts and processed for serum separation by using gel/clot activator tubes. Separated serum was analyzed by competitive ELISA. Further, statistical analysis was done by OpenEpi to check the association between BTV seroprevalence and potential risk factors. Later the BTV prevalence has been mapped in relation to different villages of both districts. Results of present study revealed that Bluetongue virus is prevalent in Faisalabad and Lahore districts with high seropositivity observed for Faisalabad district. Antibodies to BTV were detected in all studied animals irrespective of their age, sex, parity and breed. Risk factor analysis is implicating the association of BTV seroprevalence with breed, sex and age for sheep, SUMMARY 44 cattle and buffalo respectively. Further studies should be conducted to expand the geographical area for the assessment of Bluetongue prevalence and to explore the genetic diversity of Bluetongue virus. Availability: Items available for loan: UVAS Library [Call number: 2497-T] (1).

8. Nucleotide Sequence Variation In Heat Shock Protein 70-1 Gene Of Capra Aegagrus Blythi

by Fehmeeda Fatima (2014-VA-775) | Dr. Asif Nadeem | Dr. Maryam Javed | Dr. Abu Saeed Hashmi.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Heat shock protein 70 (HSP 70) plays a vital role in survival of an organism by providing cytoprotection against various kinds of stresses. Among all the HSPs present in the cell, the ubiquitous HSP 70 proteins are the most abundant and temperature sensitive. Considering the importance of HSP70-1 gene in conferring thermotolerance, present study has been designed to characterize this gene in Sindh ibex which is a wild goat species of Pakistan. The characterization of HSP70 gene might be helpful for deriving phylogenetic relationship among different species and identifying new functions among the related species. Blood/meat samples (n=25) were collected from Kirthar national park, Sindh. Standard DNA extraction method was used for DNA extraction. PCR primers were designed by Primer3 software and amplification of gene was done by Polymerase Chain Reaction. PCR product was sequenced bi-directionally by Big DyeTM Terminator on ABI 3130XL Genetic Analyzer. Multiple sequence alignment was performed for polymorphism identification. Genetic diversity was calculated by using DnaSP v.5.0. Phylogenetic analysis using the MEGA v.6.0 software package was performed and neighbor joining and UPGM trees were constructed. The results indicated that Sindh ibex HSP70.1 gene was highly similar to of domestic goat, sheep, cattle, buffalo, camel and horse which indicates their origin from a common ancestor. The results of this data might be helpful in designing effective conservation strategies for Sindh ibex. Availability: Items available for loan: UVAS Library [Call number: 2524-T] (1).

9. Molecular Exploration Of Zbed6 Gene For Growth Trait In Lohi Sheep

by Usman Sagheer (2014-VA-03) | Dr. Maryam Javed | Dr. Akhtar Ali | Prof. Dr. Tahir Yaqub.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: ZBED6 gene is a central transcription factor. It is as a repressor of IGF2 (insulin-like growth factor II) interpretation in skeletal muscle myogenesis and development. It is essentially included in organism development, signaling, cell to cell collaboration, hepatic fibrosis, clathrin intervened endocytosis and tight intersection signaling falls. Chromatin immune precipitation (ChIP) sequencing utilizing C2C12 cells recognized around 2,500 ZBED6 binding locations in the genome, and the derived accord theme gave an immaculate match with the set up tying site in IGF2. Silencing of ZBED6 in myoblast cells influences IGF2 expression, wound healing, cell proliferation and myotube arrangement. Genes connected with ZBED6 binding sites demonstrated a very huge advancement for certain Gene Ontology groupings, including improvement and transcriptional regulation. Forty two blood samples were collected. DNA extraction was done by using organic extraction method. Primers for PCR amplification designed using Primer3 software. PCR products were sequenced and then analyzed by using BioEdit software. Expasy translational tool for translation and POPGENE 32 software for analysis of population genetics at all the loci were used. Using this software the overall allele frequency, heterozygosity, probability using Chi-square test and Likelihood ratio test and Hardy-Weinberg equilibrium, genotype distribution at all SNP position, summary of genetic variation statistics for all loci and association were calculated. After this, for the association one way ANOVA was performed. Single nucleotide polymorphism within ZBED6 could be potential candidate gene to be serving as genetic marker for the selection of animals with higher tendencies towards weight gain. Availability: Items available for loan: UVAS Library [Call number: 2539-T] (1).

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

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

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

11. Molecular Investigation Of Low Density Lipoprotein Receptor Gene Causing Familial Hypercholesterolemia And Its Evolutionary Relationship With Pan Troglodytes

by Rida Zainab (2014-VA-808) | Dr. Maryam Javed | Dr. Asif Nadeem | Prof. Dr. Tahir Yaqub.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Familial Hypercholesterolemia (FH) phenotype is related to improper metabolism of low density lipoproteins due to mutations in Low-density lipoprotein receptor (LDLR) gene with increased risk of ischemic heart disease. Genetic variants in LDLR gene are associated with defective catabolism of cholesterol effecting lipid metabolism which results in familial hypercholesterolemia. It occurs in both forms: Homozygous Familial Hypercholesterolemia and Heterozygous Familial Hypercholesterolemia. Patients having high cholesterol were identified by observing the values of their serum lipid profile test reports. Their detailed history was taken and blood samples from the identified patients of familial hypercholesterolemia were collected. DNA extraction was done by Organic method. Primers were synthesized and PCR was conducted using optimized recipe and conditions. PCR products were sequenced. Sequenced data was analyzed using Chromas or BioEdit software. BLAST was performed and sequences were aligned individually by comparing it to the reference sequence. This showed difference in any specific position of a mutated sequence against the reference sequence. CLUSTALW aligned all the sequences together in one time. Sequences were compared with reference sequence to detect the presence of any mutation or SNPs. SNPs were identified manually and the peaks were observed in order to determine if the genotype is heterozygous or homozygous. Statistical Analysis was done and any amino acid change due to the observed SNPs was determined by using Expasy Translate Tool. It was found that both the SNPs showed amino acid changes. In the end, homology analysis was done which showed that Homo sapiens had their LDLR gene closest to that of Gorilla gorilla gorilla. Availability: Items available for loan: UVAS Library [Call number: 2551-T] (1).

12. Prevalence Of H9n2 In Biotic And Abiotic Factors Post Avian Infleunza Outbreak In Different Districts Of Punjab

by Iqra Mahfooz (2010-VA-301) | Prof. Dr. Tahir Yaqub | Dr. Aamir Ghafoor | Dr. Maryam Javed.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Avian influenza H9N2 is not only a potential threat to the poultry industry but it is also a disease of zoonotic importance. In the past it caused very high rate of mortality in the poultry and cause huge economic loses. Present investigations were aimed to find out the uncommon resting points of avian influenza H9N2 virus in the environment. This virus is very dangerous for the poultry industry of the country so it is important to find out the hiding places of virus so that we can stop or control the future outbreaks of virus in the poultry and minimize the economic loss of the country. To rule out the above condition a total of 150 biotic and 200 abiotic samples were collected. Refrigerated samples were processed in the Influenza laboratory. Virus isolation and propagation was done through egg inoculation technique. Presence of virus was confirmed by using Polymerase chain reaction (PCR). The biotic samples (11/150) 7.0 percent reacted positive to HA, HI and also confirmed by PCR. All the abiotic samples were found negative for any evidence of presence of avian influenza virus. This study helped us in understanding the natural reservoirs of avian influenza virus. This study design revealed the hibernation of H9N2 virus in the apparently healthy flock production of broilers. Availability: Items available for loan: UVAS Library [Call number: 2563-T] (1).

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

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

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

14. Study Of Tyrosine Hydroxylase (Th) Gene Sequence Variations In Association With Δ9-Tetrahydrocannabinol (Thc) Dependence

by Ali Raza (2015-VA-446) | Dr. Maryam Javed | Dr. Asif Nadeem | Prof. Dr. Tahir Yaqub.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Anti-Social Personality Disorder (ASPD) is ability of an individual to adopt social norms. These ASPDs are driving force in majority of criminal activities. Dopamine being important neurotranmiter of nervous system controls major behavioral traits. Low level of dopamine can be causative factor for an individual to start drug abuse to restore it because majority of drug are proven to enhance dopamine production. In this study genetic exploration of genes coding for dopamine producing enzymes. Tyrosine Hydroxylase (TH) gene was selected and its two regions (Intron 1 and exon 3) were amplified and analyzed through Sanger’s sequencing method followed by statistical analysis. Total five SNP were recorded at locus TH1, TH2, TH3, TH4, and TH5. One insertion, three transversion and only one mutation was transition. No exonic mutation was recorded hence no change in protein structure was found. Mutations at TH1 and TH4 were found to be highly associated with addiction. Mutant “B” allele were also present but still wild “A” was most common allele in our population. TH1, TH2, TH4 have positive correlation with addiction, TH3 is correlated with Nicotine and TH5 shown protective role against nicotine. There are few genetic changes in our population that can be associated with drug addiction statistically but still there prevalence in our gene pool is very low. We can conclude on the basis of these findings that drug addiction in our population is more likely a social issue rather than genetical. Limitations of our research was sample size. There are further possibilities for this project to investigate on mRNA level. Advanced neurobiological techniques can also be applied on subjects to analyze dopamine level in different brain regions. For genetic studies epistatic role of few other genes can also be considered for validation. Availability: Items available for loan: UVAS Library [Call number: 2858-T] (1).

15. Comparative Efficacy Of Pre And Post Exposure Prophylaxis Using Indigenous Rabies Vaccine By Im Route

by Kaneez Fatima (2010-VA-202) | Prof. Dr. Tahir Yaqub | Dr. Aamir Ghafoor Bajwa | Dr. Maryam Javed.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2017Dissertation note: Rabies is a viral zoonosis that is known to be present in more than 150 countries, including Pakistan. It is a very serious health problem especially in countries with limited resources and poor awareness. It has significant economic impact and almost 100 % mortality if not properly managed.Dogs are responsible for up to 99% of all rabies transmissions to humans. Rabies is vaccine preventable viral disease. The vaccine is very expensive and a significant factor in patient’s compliance in Pakistan especially in rural areas where the main problem exist i.e. more stray dogs and increased probability of being bitten. Availability of a cheap indigenously produced effective vaccine can be very helpful in reducing the cost and overall improvement of the rabies problem in Pakistan. We randomly selected a total of 50 patients visiting IPH. Among them 10 of pre-exposure prophylaxis and 40 for post-exposure prophylaxis. Twenty patients of the post-exposure group were children and twenty were adults. The NIH-anti rabies vaccine was administered intramuscularly to the persons visiting the IPH for pre and post exposure prophylaxis using Essen regimen. For pre exposure patients three doses on day 0,7,28 was given and for post exposure patients five doses on day 0, 3, 7, 14, and 28 was administered. The 3ml blood was collected on day 0, 28, 60 and 90 following vaccination. Serum was examined by ELISA Kit (Bio Rad Platelia rabies II Kit) for protective antibody titer. Pakistan is importing anti-rabies vaccine which is much costly, and sometimes unavailability is a serious concern for patients. In the present study we concluded that indigenous rabies vaccine was very effective and protective levels of rabies antibody titers was detected following vaccination in all patients of the study. By widespread utilization of this vaccine we can reduce demand of imported vaccine, thus lessen the economic burden. Availability: Items available for loan: UVAS Library [Call number: 2875-T] (1).

16. A Thesis Submitted In The Partial Fulfillment Of The Requirements For The Degree

by Ayesha Saddiqa (2011-VA-367) | Dr. Asif Nadeem | Dr. Maryam Javed | Dr. Muhammad Zubair Shabbir.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2017Dissertation note: AIDS epidemic is increasing rapidly in the Eastern Mediterranean Region. Quoting fresh authorized figures collected by the Punjab health department were 97,000 to 125,000 HIV positive people in Pakistan. Number of patients with HIV/AIDS rapidly increased in Punjab. 310 HIV/AIDS cases (35 women and 13 children) have been stated in Punjab in 2014. CCR5 gene is associated HIV infection. Mutation in this gene delayed the progression towards AIDS. In this study blood samples were collected from the laboratory of Punjab Aids Control Program (PACP), primary and secondary health care department, Government of Punjab. Genomic DNA was extracted by using the using FavorPrepTM Blood/Cultured Cell Genomic DNA Extraction Mini Kit. Specific set of primers were designed for the amplification of the targeted gene. The amplified PCR products were precipitated and sequenced for the identification of polymorphisms. Bidirectional sequencing was done for result confirmation. Alignments of sequences were done with the help of NCBI BLAST. Chromas software, Clustal W, UCSC, Bio Edit and SNPedia and Mega 6.0 was used to compile this study. CCR5 32 base pairs allele deletion was found absent in all HIV positive and negative individuals. So, susceptibility of human immuno-deficiency virus type one infection is high in Pakistani population. Genomic comparison was done with non-human primates. Alignment result showed human CCR5 gene homology, 95%, 99%, 94% and 94% with Maccaca mulata (Rhesus Monkey), Pan troglodytes (chimpanzee), Cercocebus atys (sooty mangabey) and Rhinopithecus bieti (black snub-nosed monkey) respectively. So, this homology analysis showed that these non-human primates can be used for development of therapeutic strategies related to human immune deficiency virus. Availability: Items available for loan: UVAS Library [Call number: 2872-T] (1).

17. Association Of Tryptophan Hydroxylase 2 Gene Polymorphisms With Risk Of Depression In Homo Sapiens And Equus Caballus

by Sher Sarmad (2015-VA-1062) | Dr. Asif Nadeem | Dr. Maryam Javed | Prof. Dr. TahirYaqub.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: In the biosynthetic pathway for brain serotonin, Tryptophan hydroxylase-2 (TPH2) acts as the rate-limiting enzyme. It is a key element in maintaining adequate serotonin neurotransmission in the central neuron system (CNS). It is broadly discussed as an important candidate gene in multiple psychiatric disorders, especially suicidal behavior and depression. A relationship between TPH2 and major depressive disorder (MDD) has been reported by multiple gene-disease association studies in different populations. Horse can be employed as a valuable candidate for an animal model of depression because it shares environmental factors which are known to cause depression in humans.The hypothesis of this study was that, there is association between TPH2 gene polymorphisms and risk of developing MDD.Blood was collected from each participant.Human (Experimental and Control group) and Horse (Experimental and Control group).DNA wasextracted usingthe standard Phenol Chloroform Isoamyl alcohol (PCI) protocol. Specific set of primers were designed for the amplification of TPH2 gene exons and partial region of the introns. The amplified PCR products was precipitated and sequenced for the identification of variants. For sequence data analysis Chromas Software(2.1) was used along with BLAST available at NCBI and Clustal W program. Multiple alignments were performed for polymorphism identification and association of identified polymorphisms was performed using SPSS.The significant association of the variant rs7305155 with Major Depressive Disorder indicates that TPH2 has a role in disease etiology.Understanding the extent of the role different genes play in MDDmay help in tailoring medication. “Gene-Response to drug” association studies have also shown that patients with certain polymorphisms might respond better to certain class of drugs. It is useful to know which variants are prevalent in our population. Availability: Items available for loan: UVAS Library [Call number: 2939-T] (1).



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