1.
Genetic Study Of Myp6, Mpy7, And Myp8, Loci Of Myopia In Punjabi Families
by Maria Arshad | Prof.Dr.Masroor Elahi Babar | Dr. Abu Saeed | Dr. Ali Raza Awan.
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Publisher: 2010Dissertation note: Myopia is a refractive abnormality of the eye in which the parallel light rays from an object at optical infinity are focused by the eye in front of the retina rather than on it. It may be syndromic or non-syndromic. An extreme genetic heterogeneity is associated with this disorder. This is the first experimental study on Myopia in Pakistan. So, investigating the loci of myopia here is very important because this disease is spreading day by day with prevalence rate of 36.5%.
Microsatellite markers have been proved as an efficient and powerful tool for discovering any diseased locus. So a panel of these markers was used in this study. Blood samples of various myopic families were collected from various areas of Punjab and their DNA was extracted with the standard protocol. The amplification of DNA was done with primers of microsatellite markers belonging to the loci MYP6, MYP7 and MYP8. Genotyping was done for linkage analysis through PAGE. Haplotypes were made manually by observing the alleles of all the individuals on the gel. The results showed potential linkage against MYP7 locus for the family Myo-3 with autosomal dominant mode of inheritance. This family belongs to the caste "Khawaja" and was enrolled from PCSIR Phase II, Lahore, Punjab. All the affected individuals carried the same allele that was not present in the normal subject. Later the LOD Score for this family was calculated and maximum LOD score came out to be 0.0803 at the marker D11S904 that showed very low percentage of linkage. This can be confirmed by extending the family by further sampling.
Availability: Items available for loan: UVAS Library [Call number: 1157,T] (1).
2.
Clinical And Genetic Study Of Myopia In Myopic Families From Lahore.
by Nabeeha Moeen | Prof.Dr.Masroor Elahi Babar | Dr. Ali raza awan | Dr.Aftab ahmad.
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Publisher: 2010Dissertation note: Myopia is described as the common cause of impaired vision and visual disability. In this disease the image is not focused sharply on the retina causing a blur vision to be formed and this condition of eye is referred as myopia. It is highly prevalent eye disease with its prevalence estimated to be I trillion throughout the world and approximately four billion in Pakistan. It is multi factorial disease with 19 loci identified up to date. Five myopic families were identified and selected for this study from different areas of Lahore. Linkage analysis of these families was done by MYP3, MYP4 and MYP5 loci (each consisting of a set of 3 microsatellite markers) of myopia that were selected from the panel of 19 loci. A total number of 9 microsatellite markers were used to analyze 24 samples from five families. After DNA extraction and PCR amplification, linkage analysis was carried out by genotyping through PAGE and haplotypes were constructed for the families.
Through the haplotype analysis of pedigree it was found that none of the families was found linked on any of the loci. The comparison of linkage analysis past studies with this study yielded no evidence for the presence of linkage in any of the family genotypes on the three loci. Also the LOD score calculation suggested that as all the pedigrees were found to be unlinked, the LOD score values calculated was less than 1 which suggests that markers also do not support the linkage. This may be due to the less availability of normal samples and total number of affected samples.
Moreover according to clinical factors, the individuals selected had low cylindrical component which suggest that these individuals are having simple to moderate myopia. Whereas, increase in spherical component with age shifts the lens more towards positive value (hyperopia) was also observed.
It is concluded from this study that no linkage was identified in any of the family. Both clinical and genetic factors are involved in development of myopia. Further detail study on the loci of myopia is required especially focusing the families with consanguineous marriages. Because in such families the probability of presence of linkage is more as the chances of transmission of disease allele are more in cousin marriages. From the presence of unlinked pedigrees it can also be proposed that any novel locus is present and through the identification of this novel locus, a novel gene can also be identified. Moreover, there is a probability that through genome wide screening, any other loci on any other families of Lahore may show an inherited pattern.
Availability: Items available for loan: UVAS Library [Call number: 1158,T] (1).
3.
Linkage Analysis Of Myp12 And Myp14 In Families From Lahore
by Maryam Zahra | Prof.Dr.Masroor Elahi Babar | Dr. Ali Raza Awan | Prof. Dr.
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Publisher: 2010Dissertation note: Myopia is one of the most common refractive errors of the eye worldwide that can effect clarity of vision, limit occupational choices and contribute to increased risk to vision threatening conditions. Six families of different casts were enrolled from Lahore (Punjab). Total of six autosomal dominant families were screened for linkage to the known nonsyndromic autosomal dominant and QTL myopia locus, MYP12 and MYP14 respectively. 5mL blood sample were collected aseptically in a 5Oml falcon tubes containing EDTA. DNA extraction was done by inorganic method. Three markers for each locus were selected from literature and redesigned by using 'primer 3' software. These markers were optimized for their annealing temperature and specific concentration of PCR ingredients by gradient PCR. After that all of the markers were amplified separately on genomic DNA samples of each family. PCR products of each of the marker were run on 1.2 % agarose gel along with 50 base pair ladder to visualize the bands of amplified products at 110 volts for 30 minutes. Linkage analyses were carried out by genotyping through PAGE unit of Major Science, model no. MV-2ODSYS. PCR products were run on Polyacrylamide Gel Electrophoresis (PAGE) to examine amplified bands of microsatellites. A standard 5Obp DNA ladder was run along with the sample PCR products as a reference. By reading the alleles appeared on gel haplotypes were constructed for each member of these families. The overall results of this study did not show evidence for linkage of myopia in thee families to the selected loci MYP12 and MYP14 on chromosomes 2 and 1 respectively. It might be possible any other identified locus or any new locus involved in this population of Pakistan. The findings represented here do not represent the conclusion of this study but do provide ongoing data for further investigation into the exact gentic causes of mypia in Pakistan.
Availability: Items available for loan: UVAS Library [Call number: 1216,T] (1).
4.
Study Of Genetic Association Of Pax Gene To Waardenburg Syndrome In Pakistani Patients
by Huma Tabassam | Dr. Ali Raza Awan | Miss. Sehtish Faryal | Mrs. Shagufta.
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Publisher: 2011Dissertation note: Waardenburg syndrome is an auditory-pigmentary disorder that combines clinical features of pigmentary abnormalities of the skin, hair and irides, sensorineural hearing loss, and Hirschsprung disease. Patients with WS have been shown to have mutations in the PAX gene as well as in other genes. In the present study, the locus specific polymorphisms of human PAX gene isolated from healthy and diseased Pakistani individuals was investigated for genetic association of the polymorphism with the Waardenburg syndrome. Mutation in PAX gene was identified from Pakistani patients with Waardenburg syndrome.
For the purpose, blood samples of patients suffering from Waardenburg syndrome were collected from different hospitals. DNA was isolated from WBCs suspended in the preserved samples using standard organic DNA extraction protocol. Primers were designed using Primer3 programme. PCR conditions were optimized and mutation discovery was performed on all DNA samples. Analysis of the sequences and mutations was done with the help of appropriate bioinformatics softwares. Analysis of the variable sites revealed T?C transitions, (mutations) at position number 244 was found in exon 2 of the gene. All 17 patients exhibit this mutation at the same position. Results of normal patients where there was no change found and PAX3 gene is not mentioned as it was not significant to mention. PAX gene has not been studied in the Pakistani patients earlier. The mutational investigation and association study will help to understand the genetic basis of the Waardenburg syndrome not only in Pakistan but it will also contribute to the global efforts to understand the human genetics.
Availability: Items available for loan: UVAS Library [Call number: 1359,T] (1).
5.
Association Of Genetic Polymarphism Of Cyp 2D6 Gene With Generalized Tonic Clonic Seizures In Pakistani Ptients
by Rana Manzoor Ahmad | Dr. Ali Raza Awan | Prof. Dr.Masroor Elahi Babar.
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Publisher: 2011Dissertation note: Epilepsy is chronic neurological disorder in which hyperexcitibilty of neurons cause seizures. It is a serious disorder as there is an association between the increased mortality and epilepsy. The etiology of epilepsy can be genetic. There are two main types of epilepsy, partial and generalized. These two types are further categorized into different type of seizures. Its prevalence is more in developing countries like Pakistan. The generalized tonic clonic seizure (GTCS) is a type of epilepsy prevelant in Pakistan. Cytochrome P450 (CYP2D6) enzyme has been reported to be associated with GTCS. CYP2D6 is encoded by a 4.6 kb gene named as CYP2D6. This is a highly polymorphic gene having 09 exons.
In this study CYP2D6 gene (exon 1-5) was characterized for polymorphism and the polymorphism was evaluated for asssociation with GTCS in Pakistani patients. Patient data and blood samples of different epilepsy patients were collected. DNA was isolated by inorganic method. PCR amplification was used for amplification of CYP2D6 (exon 1-5) and sequencing was performed on ABI 3130 XL Genetic analyzer. Two mutations, 214 G>C and 232 G>C in intron1 of CYP2D6 gene have been found. These mutations were only found in Pakistani patients suffering with GTCS. Absence of these mutations in 10 healthy individuals (control group) confirmed association of these mutations with GTCS. This outcome of study will help to add information in international gene data. The mutations found in this study will also lead to gene therapy of GTCS, genetic counseling and develop prenatal diagonastic tests.
Availability: Items available for loan: UVAS Library [Call number: 1390,T] (1).
6.
Identification Of Single Nucleotide Polymorphisms In Olri Gene And Its Association With Milk Composition In Cattle Breeds in Pakistan
by Nusrat Majeed | Prof.Dr.Masroor Elahi Babar | Dr. Ali Raza Awan | Prof. Dr. Abu.
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Publisher: 2011Dissertation note: Pakistan being agriculture based country has a great potential in livestock sector. it plays
an important role in the economy of the country. Milk is a balanced diet because it
contains all the essential nutrients like carbohydrates; fats, proteins, vitamins, minerals
and enzymes required for health. In these milk nutrients fat is second most important
component of milk. Primary component of milk fat is triglycerides (triacylglycerols or
TAG) a typical storage form of lipids. OLRI is the major protein that binds, internalizes
and degrades oxidized low density lipoprotein. Ol.Rl as a protein important for oLDL
metabol ism may contribute to these effects. Oxidized fat inhibits the expression of
lipoprotein lipase and fatty acid transporter genes that causes a reduced uptake of fatty
acids into mammary glands. As a result the concentration of triacylglycerols in milk is
reduced. OLRl as a protein important for oLOL metabolism may contribute to these
effects. OLR J identified as a functional and positional candidate gene product for milk fat
percentage and milk fat yield. Polymorphism in Ol.Rl gene increases the milk fat
percentage. If polymorphism in Ol.Rl gene is found in Pakistani Sahiwal and Ohanni
cattle breeds these were help to screen the animals at younger age and also be used to
characterize the different cattle breeds as a milk fat production marker. Unrelated animals
were selected for this study. Blood samples were collected from different Go\'1. livestock
farms/experimental stations. DNA was extracted by organic method. Primers were
designed by using Primer3 software. After DNA amplification by polymerase chain
reaction. peR product was sequenced. Sequencing results of the full length OLR I gene
were analyzed by alignment of sequences with the help of Blast2sequence software.
Novel S Ps were identified. Total of twenty one SNPs were identified in the samples of
all breeds that were confirmed at population level by sequencing of each sample. Then
total twenty one SNPs were found in all breeds. Most of SNPs were found in intronic
regions. In Sahiwal only 5 SNPs are exonic and one of them is common in both Sahiwal
and Red Sindhi breeds. Out of total five exonic SNPs, only two were found synonymous
and rest of the three exonic SNPs were found to be non synonymous. while all other
SNPs are intronic in nature. An intronic SNP is also common between Sahiwal and
Dhanni breed. In Red Sindhi a SNP was observed at 3'UTR. Results were analyzed by.
using the statistical software POPgene32. The mean value of expected heterozygosity that
is 0.3595 is greater than the mean of observed heterozygosity that is 0.0300 that shows
low variabil ity in breeds. The mean value for Shannon's Information index (1*) is 0.5421.
The aim of research is to identify gene that underlie the genetic variation in bovine milk.
fat composition.
Availability: Items available for loan: UVAS Library [Call number: 1396,T] (1).
7.
Association Of Katg Gene With Isoniazid Resistance In Multiple Drug Resistant Tuberculosis
by Farouk Qamar Malik | Dr. Ali Raza Awan | Prof. Dr.Masroor Elahi Babar.
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drama
Publisher: 2012Dissertation note: TB has been announced as a global emerg~ncy of this millennium. It is one of the leading causes of death among adults due to a single infectious agent. Pakistan is sixth among the twenty two Eastern Mediterranean Region countries with the highest burden of disease which is approximately 181 per 100,000. The emergence of drug resistant MTB poses a serious threat to the ongoing efforts to control the disease epidemic. Drug resistance to the first line drugs such as INH and RIF needs to be investigated. In this respect the role of various genes conferring resistance should be studied to find better treatment alternatives. The current practice of drug sensitivity testing requiring approximately three weeks (total turnaround time for MTB culture and sensitivity is around 90 days) is time consuming and a major cause of treatment delay. In this research sputum samples were collected in wide mouth transparent containers from suspected TB patients. After decontamination samples were inoculated onto LJ medium. The colonies grown on the slopes were identified as MTB by standard biochemical test. Isolates were tested on LJ medium for in vitro DST (Drug Sensitivity Testing). MDR was described as resistance to INH and RIF with or without resistance to other drugs. DNA was extracted from the grown samples using kit method. After extraction of DNA, the region from base 2714 to 3232 of katG gene was amplified through peR and the amplified products were sequenced. Analysis of the DNA sequences and mutations was done with the help of BLAST - alignment software. A total of 24 MDR MTB samples were sequenced. Sequence analysis revealed the reported mutation Ser - Thr in katG codon 315 in five samples (21 percent of the total sample size). In this study, an authentic molecular analysis (test) was developed and validated for identification of INH resistant strains in Pakistani population. By studying genetic mutations in katG gene and its association With INH resistance, an alternative can be provided whereby specimens can be tested for these mutations and timely decisions taken. This will not only save the patients from unnecessary treatment delays but will also prevent the administration of drugs to which MTB is resistant and in the long run decrease drug resistance and disease burden.
Availability: Items available for loan: UVAS Library [Call number: 1402,T] (1).
8.
Mutation Screen Of "Gamma-Aminobutyric Acid (Gaba)-A Receptor, Gamma 2" In Punjab Population
by Muhammad Javed Iqbal | Dr. Muhammad Wasim | Dr. Abu Saeed | Dr. Ali Raza Awan.
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drama
Publisher: 2012Dissertation note: Epilepsy is a formidable form of neural disorder that can impose its long lasting effect on person's life and development. To date, it lacks any effective therapy and is multistep disease strengthened by an overwhelming number of genetic and epigenetic mechanisms that streamline epileptic attacks. This particular study encompasses two major types of epilepsy, CAE and GTCS by targeting a GABRG2 gene. Mutation analysis of the coding exons (exon 3, 5 and 9) was performed by direct sequencing of GABRG2 in order to sought out complex biological entities in both types of epilepsies. GABRG2 is a molecule that has recently been characterized as the culprit for epileptic seizures onset. GABRG2 encodes GABA receptor that is fundamental inhibitory neurotransmitter in mammalian brain and is a ligand-gated chloride channels. This ligand-receptor coupling results in the inward shuttling of chloride ions through the channels and this hyperpolarizes the neurons, which induce the inhibitory effect of neurotransmitters. Direct sequencing of candidate gene "GABRG2" traced out a single polymorphic site in the exon 3 of the CAE as well as GTCS cases. However, this single nucleotide alteration is more commonly identified in childhood absence epilepsy patients as compared to the generalized cases. Silent mutation was identified at locus 27909 C>T of 46.66% of the total screened or analyzed cases.
Availability: Items available for loan: UVAS Library [Call number: 1500,T] (1).
9.
Dna Barcoding Of Pakistan Avian Families (Sturnidae & Columbidae)
by Emma Umar | Dr. Ali Raza Awan | Dr. Muhammad | Ms. Saeeda Kalsoom.
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Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1515,T] (1).
10.
Identification Of Polymorphism Of Cyp2D6 Gene In Pakistani Patients Of Epilepsy
by Sana Kulsoom | Prof. Dr. Masroor Ellahi Babar | Dr. Ali Raza Awan | Mr. Zahid.
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drama
Publisher: 2011Dissertation note: Epilepsy is a neurological disorder which causes seizures by sudden burst of hyperactivity in the brain. It is a global health problem and 50 million people have epilepsy all over the world. The majority of epilepsy patients can pass a good life if they receive proper treatment. Various types of epilepsy are present in Pakistan but Generalized tonic-clonic seizure (GTCS) is the most frequent type of epilepsy present in Pakistani patients. GTCS is a type of seizers involving the entire brain. It is also called a grand mal seizure. CYP2D6 is a gene associated with GTCS.
CYP2D6 is a 4.6 kb gene which encodes the protein cytochrome P450, a metabolizing enzyme. CYP2D6 is a highly polymorphic gene with 09 exons. Patient data and blood samples of different epilepsy patient were collected. DNA was isolated by Organic method. PCR amplification was performed for CYP2D6 gene (exon 6-9) and sequencing was performed on ABI 3130 XL Genetic analyzer and the results were analyzed. In this way the mutations, 2850 C>T in exon 6 of CYP2D6 gene have been caricaturized in Pakistani patients of GTCS encoding Cyctine in place of Argine at 296 in CYP2D6 enzyme. This change may be associated with GTCS in Pakistani Patients. Above study will help medical scientists in Molecular diagnosis, Genetic counseling and Gene therapy.
Availability: Items available for loan: UVAS Library [Call number: 1519,T] (1).
11.
Genetic Characterization Of Pakistani Lathy Pigeons Using D-Loop, Cyt B And 16S Rrna Genes As Genetic Marker
by Muhammad Umair Latif | Ms. Sehrish Firyal | Dr. Ali Raza Awan | Dr. Muhammad.
Material type: ; Format:
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not fiction
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1399,T] (1).
12.
Molecular Charaterization Of Ampk Gene Of Pakistan Buffalo
by Waqas Ahmed Khan | Prof. Dr. Masroor Ellahi Babar | Dr. Aftab | Dr. Ali Raza Awan.
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Publisher: 2011Dissertation note: Pakistan is an agriculture country and its economy is mainly dependent on agriculture, agriculture products. Livestock has been playing an important role in the economy of the country. Livestock sector contributed approximately 51.8 percent of the agriculture value added and 11.3 percent to national GDP. Buffalo which is known as black gold of Pakistan is famous for its largest milk production in the world. A better understanding of the genetic control of energy metabolism in farm animals can have far-reaching implications for molecular breeding programs. It can allow the implementation of knowledge-based breeding to increase feed efficiency and to improve meat quality. In addition, because of the high degree of evolutionary conservation of these genes, the information gained about the genetic control of animal nutrition can be extrapolated back to questions about human nutritional genomics and disease. This study was performed to discover the single nucleotide polymorphism at AMP-activated protein kinase (AMPK) gene in Nilli Ravi and Kundi Buffalo and their possible association with milk production. As AMPK is a sensor of energy metabolism so genetic variations in AMPK gene may also have effect the feed utilizing efficiency of animals. Buffalo is popular for utilizing low quality roughages in a better way. Buffaloes are popular in the world for high fat content and low cholesterol content as compare to cattle. A total of 128 single nucleotide polymorphisms were discovered at AMPK gene in Nilli-Ravi and Kundi Buffalo. Out of which 10 are in exonic region and 118 are in Intronic region. Most of the SNPs are Intronic it also shows that AMPK is highly conserved as it has been shown by many studies. The Intronic SNPs may have role in regulation of AMPK gene. Forty-six SNPs were discovered in Intronic region of A1 subunit of AMPK gene. Out of these 46 SNPs. Forty-four SNPs are same in both Nilli-Ravi&Kundi buffalo.
Two SNPs found at position 11908 and 12217 was present only in Kundi buffalo. These two SNPs can be used for breed characterization of Nilli-Ravi&Kundi buffalo. The numbers of SNPs discovered in exonic region are 6. These all SNPs are non-synonymous mutations and changes amino acids at position 23333 from Histidine>Tyrosine, at 23387 from Glutamic acid>Lysine, at 23402 from Valine>Isoleucine, at 23426 from Ser>Pro, at 23489 from Stop codon>Arg and at 23612 from Ala>Thr. Forty SNPs were discovered in Intronic region of A2 subunit of AMPK gene. Out of these 43 SNPs 28 are same in both Nilli-Ravi & Kundi buffalo. SNPs at positions 71371, 71382, 71383, 71396, 71558, 42736, 42766, 42881, 41661, 41900 and 42021 are only present in Kundi buffalo while SNPs at position 70900, 71613, 42935 and 42944 are present only in Nilli-Ravi buffalo. These SNPs can also be used for breed characterization of Nilli-Ravi and Kundi buffalo. The B1 subunit of AMPK gene has 21 SNPs in Intronic region, which is common, both in Nilli-Ravi and Kundi buffalo. These polymorphisms may have role in regulation of AMPK gene. The SNPs found in exonic region are 3 which are all non-synonymous mutations and changes amino acids at position 4362 from Histidine>Tyrosine and at positions 8193, 8195 from Glycine>Serine. All exonic SNPs are non-synonymous mutations, which show that it will change the function of protein and might be associated with milk production and feeding efficiency in Nilli-Ravi & Kundi buffalo. This study is an example of candidate gene approach to find some novel variations at population level. It is the first study conducted for Molecular Characterization of AMPK gene in Buffalo. The only way to associate these polymorphisms to the trait under consideration (energy metabolism) by back tracing the sampling groups. This study is first in finding some molecular markers for energy metabolism in Nilli-Ravi and Kundi buffalo that can be used for future selection and breeding programs. More the population will be diversified for the trait and showing trends of heterozygosity, better will be the chances of selection of animals with suitable genetic makeup.
Availability: Items available for loan: UVAS Library [Call number: 1796,T] (1).
13.
Genetic And Evolutionary Characterization Of Pakistani Pigeons And Parrots Through Mitochondrial D-
by Sehrish firyal | Dr. Ali raza awan | Prof, Dr. Aftab | Prof, Dr. Tahir yaqub.
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Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1873,T] (1).
14.
Mitochondrial Nd2 Gene Based Molecular Classification Of Various Pakistani Domestic Pigeons
by Muddasar Saeed khan | Dr. Sehrish firyal | Dr. Ali raza awan | DR.Muhammad.
Material type: ; Format:
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not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1983,T] (1).
15.
Molecular Phylogeny And Diversity Analysis Of Gazella Bennettii (Chinkara) Deer Of Pakistan Using D- Loop Region Of Mitochondrial DNA
by Maleekah zaheer | Dr. Muhammad yasir zahoor | DR. Ali raza awan | Miss. Faiza.
Material type: ; Format:
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not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1985,T] (1).
16.
Differential Expression And Mutation Analysis Of Heat Shock Proteins (Hsps) And Tumor Suppressor Gene (P53) In Differemt Cancer Types of Pakistani Dogs and Cats
by Rashid Saif | Dr. Muhammad Wasim | Dr. Ali Raza Awan | Dr. Muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2170,T] (1).
17.
Polymorphisms Of Bovine Tumor Necrosis Factor Alpha Gene And Its Association With Mastitis In Sahiwal Cows
by Huma Sattar (2013-VA-03) | Dr. Sehrish Firyal | Dr. Ali Raza Awan | Dr. Muhammad Tayyab.
Material type: ; Literary form:
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Publisher: 2015Dissertation note: Mastitis is one of the shocking maladies of milch animals causing high production losses to livestock industry of Pakistan (Kenyanjui et al. 2011). It is an inflammatory condition of udder; represent a major problem in dairy cow management. It is one of the most common and frequent disease of dairy industry. Producers suffer a huge loss due to veterinary treatment costs and necessary culling of the infected animals. It negatively affects the milk production, quality of milk, and farm economics (Fourichon et al. 2005). Increasing the disease resistance among dairy cattle is therefore desirable because without controlling mastitis, the national goals of developing dairy farming on commercial and scientific lines and production of wholesome milk which conforms to the standards of WTO Accord would remain elusive.
Mastitis is inflammation of udder that caused by physiological and metabolical changes (Schalm and Noorlander 1957). There are two main types of mastitis; clinical mastitis (characterized by classical symptoms i.e., swelling of udder, redness, clumps and clots in milk etc) and sub-clinical mastitis (not show any symptoms, Milk appear normal, udder appear normal) (Schrick et al. 2001). Mastitis is ranked as a top disease of dairy herds (Rinaldi et al. 2010). This mammary gland infection caused by pathogenic micro organisms such as Staphylococcus aureus, Streptococcus uberis, and Esherichia coli in the mammary gland (Heringstad et al. 2000).
India, China and United States are the larger producer of milk and Pakistan is on forth number in milk yield. Pakistan almost produces 36.5 million tons of milk yeild per year (Cady et al. 1983).The Sahiwal breed is well known among for its superior dairy qualities (Barker et al. 1998). Both cross and pure breed Sahiwal cows have high milk production rate (Khan et al. 2013).
It is very difficult to comprehend this disease because numerous environmental and genetic factors are involved in the origin and development of mastitis (Bradley 2002; Carvajal et al. 2013). Susceptibility and resistance to mastitis is a complex trait influenced by genetic variation of animals. Among these variations, the polymorphisms in immunity genes are principal key factors in defensive mechanism of mammary gland (Ibeagha-Awemu et al. 2008).
The mammary gland tissue is protected by immune system by two defense system; innate and acquired immunity. Innate immunity response by the host is a quick response of bacterial defense system (Mesquita et al. 2012). Innate system is a rapid and effective mechanism that activated on recognition of antigen (Akira et al. 2006). Innate immune system is activated when specific pattern recognition receptors (PRR) that are present on the surfaces which are attach to the specific pathogen (Shuster et al. 1996). PRR are presnt on leucocytes in milk and on the epithelial cells lining of udder. It is reported that T- lymphocyte subset i.e., CD4+, CD8+ and ɤδT are present in infected bovine mammary glands. (Goldammer et al. 2004; Strandberg et al. 2005).
Innate defense (nonspecific) of the mammary gland is stimulated by the physical barrier such as teat end, natural killer (NK) cells, neutrophils, macrophages and certain other soluble factors. The teat cannals are considering the main line of defense. Microorganisms enter from teat canal in milk. The main roles of teat sphincter muscles are to remain orifice close so that bacteria cannot enter. This teat canal also lined with keratin, whose estrified and non estified fatty acid function as bacteriostatics that provide protection and play role to eliminate bacteria causing mastitis (Oviedo-Boyso et al. 2007).
If a pathogen is not eliminated by the physical barrier, the acquired immune system is triggered. In comparison, this system is much faster than other immune response. The memory response is significantly stronger, long durable and more efficient to kill the pathogen. The acquired immune system (memory response) have ability to differentiate self or nonself cells and produce antibodies only against antigens through membrane bound protein called major histocompatibility complex (MHC) molecules. Specific immune system activate only when antigens bind with an MHC that is present on the surface of certain cells, this process is referred as antigen presentation. Recognition of pathogenic factors for elimination is mediated by macrophages, several lymphoid, and immunoglobulins (Ig) or antibodies (Sordillo and Streicher 2002).
The most acute responding macrophages and T-cell cytokines are TNF-α, LTF, IL1, IL6, IL8, and IFN-ɤ present in intramammary infection in cows. These genes play important role in improvement of immunity to mastitis (Burton and Erskine 2003).
Tumor necrosis factor alpha is main pro-inflammatory adipokine that is part of systematic immune defense. The main function of TNF-α gene is responsible for proliferation, differentiation and activity of many immune system cells; B lymphocytes, NK (natural killer). It also induces the production and release of many other cytokines (Wojdak Maksymiec et al. 2013) and also enhances the chemotactic and phagocytic effects of immune response. TNF-α gene contains four exons and three introns that are present on chromosome BTA23q22 (Bannerman 2009; Moyes et al. 2009).
TNF-α is a member of a group of cytokines that stimulate the specific immune system. TNF consist of 212 amino acid arranged in stable homotrimers (Kriegler et al. 1988; Tang et al. 1996). The 17-kilodalton (kDa) TNF protomers are composed of two β-pleated sheets and β-strands, joined together antiparallel (Tang et al. 1996).
TNF-α is a component of natural protection systems of humans and animals. Milk gives nourishment and disease resistance to the new born. Various cellular and soluble immune components are important for protecting the mammary gland from infectious diseases like mastitis. Mastitis affects one third of all dairy cows and cost the dairy industry about 2 million dollars annually (National Mastitis Council (1996). Dairy cattle are especially susceptible to mastitis due to diminished mammary gland defense mechanisms (Sordillo and Streicher 2002).
TNF-α is not only produced by activation of macrophages, but also other cell types such as CD4+ lymphocytes, NK cells, neutrophils, mast cells, eosinophils, and neurons. Large amounts of TNF are released in response to lipopolysaccharide, other bacterial products, and Interleukin-1 (IL-1).TNF-α stimulates the proliferation, differentiation and activity of many immune system cells; B lymphocytes, NK (natural killer). TNF-α induces the release of many other cytokines (Wojdak-Maksymiec and Mikolajczyk 2012). TNF-α also enhance the chemotactic and phagocytic effects of immune response.
. The present study is designed to determine the genetic polymorphism in exon 4 of TNF-α gene of mastitic cows and its association resistance and susceptibility towards mastitis.
Availability: Items available for loan: UVAS Library [Call number: 2224-T] (1).
