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1. Parentage Analysis And Breed Characterization Of Dogs By Microsatellite Markers

by Muhammad Sajid Tahir | 5/31/2010 | Mr.Tanveer Hussain | Mrs.Saeeda.

Material type: book Book; Format: print Publisher: 1990Dissertation note: Pakistan has vast population of dogs belonging to different breeds. Most of the dogs have no pedigree record which is a great threat to conservation of different breeds. No study on DNA fingerprinting of dogs has been conducted in Pakistan. DNA fingerprinting of dogs is necessary to overcome the problems like forensic cases, sale & purchase, individual identity in case of fertilization by more than one male and ownership disputes. Microsatellite markers have been proved as an efficient and powerful tool for parentage testing and breed characterization of dogs. In this study, a panel of microsatellite markers, having high polymorphism information content (PlC) values, was developed. Blood samples were taken from cephalic vein of two breeds of dogs (German shepherd and Labrador retriever). DNA was extracted by Inorganic method. Primers of microsatellite markers were optimized for successful amplification conditions in the Bio-Rad thermocycler. Multiplex PCR was performed, for amplification of these microsatellite markers on 46 samples belonging to 20 families. Genotyping analysis was performed for the PCR products of microsatellite markers on non denaturing polyacrylamide gel. These results were analyzed statistically software "POPGENE 3.3 and POWER STAT". Allele frequency, heterozygosity, homozygosity, polymorphism information content (PlC), power of discrimination and power of exclusion of all microsatellite markers were calculated. Average power of discrimination among non parents, average hetrozygosity, average observed homozygosity and average polymorphism information content (PlC) value for all alleles was 0.809, 0.6345, 0.2913 and 0.724 respectively. Moreover combined power of exclusion reached a significant value of 0.9998. Almost all of the microsatellite markers showed significant variations in both German shepherd and Labrador retriever breeds. Microsatellite "REN41D2Ob" showed maximum variation i.e. 17 alleles and microsatellite"REN49F22b" showed the least variation among all microsatellite markers i.e. 4 alleles. Genotyping results of microsatellite markers were clearly different for two different breeds showing a distinct genetic distance between German shepherd and Labrador retriever breeds. Results of this study lead to development of a panel of microsatellite markers which can be used for parentage analysis and breed characterization of dogs. This was a preliminary study on dogs in Pakistan. This facility can be provided on commercial basis to pet owners and kennel clubs. Moreover this study can become the basis for further research investigations in canines in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 1090,T] (1).

2. Horses Parentage Analysis And Breed Characterization By Microsafellipe Markers

by Javed Iqbal | Prof.Dr.Masroor Elahi Babar | Dr.Ahmad Ali | Mrs.Saeeda Kalsoom.

Material type: book Book; Format: print Publisher: 2002Dissertation note: Horses (Equus caballus) have been considered one of the most significant domesticated animals in human use, including sports, urban and rural transportation and military logistics. Paternity confirmation and breed identification is the basic pre-requisite for rearing specific breeds and maintaining the pedigree records of horses. DNA finger printing has been successfully used for paternity and breed confirmation. Mainly DNA finger printing is done through microsettalite markers for paternity analysis and breed characterization. The aim of this study was to develop and apply a panel of microsettalite markers for paternity analysis and selected horse breeds characterization. Three horse breeds Percheron, Pak Arab and Thoroughbred were selected for this purpose. Sampling of 20 families (Foals, Mare and Stallion) of three selected horse breeds was conducted from Remount Depot Mona. The blood samples was collected in sterilized Falcon tubes each containing lOOjiL EDTA (0.2 mM). DNA was extracted from all blood samples by using inorganic method. The microsatellite markers were selected from ISAG (International Society of Animal Genetics), Indian and TKY recommended panels. The primers were designed for these microsatellite markers using primer3 free ware. All Microsatellite markers used were direpeats. Conditions for successful amplification by PCR (Polymerase Chain Reaction) were optimized and microsatellite markers were grouped into 6 multiplexes (tn and diplex). PCR products of optimized multiplexes were visualized on U V illuminator after gel electrophoresis. DNA amplification was done through PCR by using optimized multiplexes of microsatellite markers for all the samples. Polyacrylarnide Gel Electrophoresis (PAGE) was used for genotyping of amplified DNA samples. Allele sizes of all amplicons were calculated by relative flow method. Alleles for all microsatcilite markers were analyzed statistically by "POPGEN 32 and POWER STAT" software. The investigation revealed average PlC value 0.97, average observed heterozygosity 0.923 1, average observed homozygosity 0.0769, combined power of exclusion (P.E) 0.9999 and combined polymorphic loci percentage for 13 microsettalite markers was 100%. Perchron breed showed genetic identity with Pak Arab and Thoroughbred up to 0.3470 and 0.6157 respectively. Pak Arab exhibited genetic identity with Thoroughbred horse up to 0.6616 where as Perchron breed showed genetic distance with Pak Arab and Thoroughbred up to 1.0585 and 0.4850 respectively. Pak Arab exhibited genetic distances with Thoroughbred up to 0.413 1. Results of analysis were used to describe the new microsettalite markers assay for parentage confirmation and breed characterization of selected breeds (Perchron, Pak Arab and Thoroughbred) horses. 'Ihis panel of microsetallite marker was useful and reliable tool for individual identification and parentage analysis in horses. This microsettalite marker panel can be used on commercial basis in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 1092,T] (1).

3. A Study Of Plasma Homocysteine And Copper In Patients Of Coronary Artery Disease

by Umer Saeed Ansari | Prof. Dr. Ijaz Ahmad | Dr. Habib-ur-Rehman.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2010Dissertation note: The present study was carried out on 60(56 males and 4 females) stable coronary artery disease patients selected from the angiography department of Shaukat Khanum Medical & research Laboratories. Only those patients were selected as cases, who verified angiographically, as having coronary artery disease. Thirty controls were also selected from angiography department of Shaukat KhanumMedical & research Laboratories. These were the patients who on angiography were verified as having normal coronary arteries. The patients were between the ages of 30-60 years. Mean age of the cases was 43.95±5.6 years and the mean age of controls was 42.87±7.27 years (Table 1). No significant difference was found between the distribution of patients age among cases and controls. Among the cases 93% were males and 7% were females. Various risk factors which predispose to coronary artery disease were also . recorded in our study such as history of hypertension, smoking, history of hyperlipidemia, family history of coronary artery disease, obesity, serum cholesterol, serum triglyceride, serum copper and plasma Homocysteine. Regarding the history of smoking, there were 51.7% smokers among the cases. In the control group only 30% were smokers and this difference was statistically not significant (p value 0.05) (Table 3). History of hyperlipidemia was present in 17 cases and 4 controls. The family history of coronary heart disease was seen in 33 cases and 11 controls. There was no statistical difference between the distribution or these factors among cases and controls (Table 3). The cases had a mean BMI of 27.38±3.75and the controls had a mean BMI of 27.l4±5.56. In the control group 63.30/0 were overweight and obese and among the cases 71.6°1<> were overweight and obese. A number of biochemical tests including serum cholesterol, serum triglyceride, serum copper and plasma Homocysteine, were done on the study population. The mean serum cholesterol among the cases was 184.23±37.83mg/dl and in the controls it was 171.07±48.24mg/dl. No difference was found between the distribution of mean cholesterol levels in cases and controls (Fig 5). The mean triglyceride level was 207±84.71mg/dl among the cases and 160±71.27mg/dl in controls. The difference was statistically significant (Fig 6). The principal observation of this study is that mean plasma tHcy of cases was significantly higher (15.21±2.67Ilmol/l) as compared to controls (10.88±1.88Ilmol/l) (p value <0.01) (Fig 7). The other major observation was that there was a significant difference in the distribution of serum copper among cases and controls when serum copper was divided into groups (Table 8). This study observed more patients with conventional risk factors in hyperhomocysteinemic subjects (n=36) than the patients having low Homocysteine level (n=54). In spite of this no association was found between hyperhomocysteinemia and these risk factors except serum copper (p value <0.01). The mean serum copper in subjects with normal plasma Homocysteine level was 81.96~g/dl and in the patients with hyperhomcysteinemia it was 1 00.82~g/d1. A positive correlation was found between serum copper and plasma Homocysteine (r=0.44) Coronary artery disease is associated with moderate hyperhomocysteinemia in our study and it shows a positive correlation with serum copper. It does not show any association with other risk factors. Since hyperhomocysteinemia is commonly seen in our patients, it is prudent to manage these subjects with vitamin supplements and adequate nutrition. Availability: Items available for loan: UVAS Library [Call number: 1094,T] (1).

4. Breed Characterization Of Red Sindhi And Tharparkar Cattle Breeds By Mitochondrial D-Loop And Gytochrome

by Nabeela Akhtar | Mr. Tanveer Hussain | Prof. Dr. Masroor Elahi Babar.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Livestock plays an important role in economy of Pakistan. Different livestock animals used for for meat, milk, draught, and sports. The genetic data of different cattle breeds like Red Sindhi and Tharparkar is not available which needs to be established for their genetic identification, conservation and to find their genetic diversity among them. Blood samples of pure bred animals were collected from their respective home tracts. The Red Sindhi cattle samples were collected from (Barani Livestock Production Research Institute, Kherimurat, Attock, Govt. Red Sindhi Cattle Breeding Farm, Tando Muhammad Khan and Department of Livestock Management, Sindh Agriculture University, Tandojam) and Tharparkar cattle samples were collected from (Department of Livestock Management, Sindh Agriculture University, Tandojam, Tharparkar cattle Farm at Nabi Sar Road and from Tando Qaiser in Sind). DNA was extracted with the standard protocol in Molecular Biology and Genomics Laboratory of Institute of Biochemistry and Biotechnology. Specific primers were designed by using special softwares Primer 3 for mitochondrial D-loop region and Cytochrom b gene from NCBI accession no. AF492350. Then after primers optimization PCR amplification was done. Then sequencing of target fragments was carried out. Sequences were alligned with the help of software blast2sequence. Single Nucleotide Polymorphisms (SNP5) were identified and comparison of 5 mitochondrial DNA haplotypes of two cattle breeds was done. Sequences were analyzed and compared with already reported sequence of Mitochondrial DNA of Bos indicuss, Bos taurus and Bubalus bubalis available at NCBI. Phylogenetic tree was constructed using MEGA 4.1 software (http://www.megasoftware.net/MEGA4.1.html) showed that Pakistani, European and Asian cattle are genetically same but different from Buffalo. This work provided the genetic data which is very helpful for determining the genetic diversity of cattle population, breed identification, animal forensic and paternity cases and making effective breeding policies and conservational activities in future. This work is very helpful about breed characterization of two cattle breeds (Red Sindhi and Tharparker) and developing understanding about genetic architecture of cattle breeds as present study conclude that six SNPs were present in both breeds, four private to Red Sindhi and 22 were private to Tharparkar. Availability: Items available for loan: UVAS Library [Call number: 1155,T] (1).

5. 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.

Material type: book Book; Format: print 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).

6. 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.

Material type: book Book; Format: print 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).

7. Identification Of Novel Snps Of Mitochondrial D- Loop And Cytochrome B In Pakistani Goat And Sheep Breeds

by Haleema Sadia | Prof.Dr.Masroor Elahi Babar | Dr. Ahmad ali | Dr. Ali awan.

Material type: book Book; Format: print ; Nature of contents: biography; Literary form: Publisher: 2010Dissertation note: Pakistan has approximately 53.79 million goats and 26.49 million of Sheep. Goats AND Sheep are kept for milk meat and wool production and contribute significantly to the income of the farmers. Thirty recognized breeds of goats and twenty eight breeds of sheep found in Pakistan. Improvement of livestock productivity per unit animal remains the primary concern of research and development efforts. The purpose of this research work was the genetic improvement of Sheep and Goat breeds. In this present study of Goat and Sheep, Ten different breeds of Goat: Barbari, Beetal, Pahari hairy, Kamori, Damani, Khurasani, L.Hairy, Teddy, Lehri goat, Nachi and ten different breeds of Sheep: Bulkhi, Dumari, Kachi, Kaghani, Salt range, Awassi, Thalli, Lohi, Krakul, Shenwari were selected but the genetic data of different goat and sheep breeds is lacking which need to be established for their genetic characterization. Blood samples of unrelated true representatives of all breeds were collected from their respective home tract. DNA was extracted with the use of standard protocol and amplification of the mitochondrial. D-loop and Cytochrome b region was done with specific primers in Molecular Cytogenetics and Genomics Laboratory in the deptt of Molecular Biology and Biotechnology. Sequencing of amplified portion of mt.DNA D-loop and Cytochrome b was done. Sequences were analyzed with the help of software blast 2 sequence. Single Nucleotide Polymorphisms (SNPs) were identified and comparison of 50, 50 samples of Goats and Sheep of Cytochrome b gene and mitochondrial D-loop region were compared with their respective reference sequences. Genetic identity between ten goat breeds were calculated by BioEdit. 50 goat haplotypes and 49 sheep hapoltypes were identified. All haplotypes were rich in AT contents. 22 conserved region were identified which were common in Goats and Sheep (BioEdit, 7.0). Goals and Sheep sequence comparison was made by using sheep Ovis aries as a referenece. 405 varibale sites were already present in Goat and Sheep. Capra hircus (AF533441) had 41 insertions and 9 deletions with respect to Ovis aries (AFO 10406.1). Phylogenetic tree was constrncted using Mega 4.1 software showing the relationships between different haplotypes. Haplotpes of this research work were compared with some world wide haplotypes of Goat and Sheep separately. Goat showed very close relationship with haplotypes of Africa, Asia and Europe, while Sheep showed close relation with Europeaon and Asian haplotypes. Both Goat and Sheep seemed to have domestication from Asia. From analysis of Goat and Sheep with their wild reported haplotypes, it was confirmed that Goats and Sheep used in this research work were domestic and had close relation with Capra aegagrus and Capra sibrica was present at the bottom of phylogenetic tree. While Sheep (Ovis aries) showed a close relation with wild Mouflon (Ovis musimon). Availability: Items available for loan: UVAS Library [Call number: 1159,T] (1).

8. Study Of Autosomal Recessive Non Syndromic Mental Retardation Locus By Linkage Analysis

by Sajjad Ali Shah | Prof.Dr.Masroor Elahi Babar | Dr. Aftab | Mr. Tanveer Hussain.

Material type: book Book; Format: print ; Nature of contents: biography; Literary form: Publisher: 2010Dissertation note: Mental retardation (MR) is the retarded conditions of mind in which the intelligence quotient (IQ) is lower than 70, associated with a deficiency in adaptive behavior such as communication and daily living skills. Mental retardation is either the only consistent handicap (non-syndromic) or is combined with other physical and br behavioral abnormalities (syndromic). It is one of the most common disorders and it affects about 1-3% of the human population, with a proportion higher in males than females. In the present study 10 families with two or more affected individuals were selected from different areas of Malakand Division and district Mardan of Khyber Pakhtunkhwa. Family history was taken and pedigrees were made personally by visiting the families and using specially designed proformas after their consent. The blood was collected from the selected families aseptically. Then DNA was extracted by standard inorganic protocol. Short Tandem Repeat (STR) markers (D3S3630, D3S3050, D3S1620) in vicinity of MR locus (MRT2CRBN gene) were selected, optimized and amplified by Polymerase Chain Reaction. The affected families were screened for linkage to MRT2A locus using Polyacrylamide Gel Electrophoresis (PAGE). The haplotypes were then constructed to determine the linkage of families to MRT2A locus. Out often selected families two families (MR-02 and MR-07) showed linkage to autosomal recessive nonsyndromic mental retardation locus MRT2A. This is the first report of MRT2A phenotype linkage in families from Malakand Division where consanguineous marriages are very common. Further study is needed to explore the other linkages in mentally retarded families in local population. The present study will help us to determine the genetics basis of mental retardation in affected families of Pakistan. It will also help us to screen out carrier individuals in our population that would help to develop genetic counseling strategies to prevent the progression of mental retardation in the country. Availability: Items available for loan: UVAS Library [Call number: 1162,T] (1).

9. Dna Typing Of Pakistani Cattle Breeds (Tharparkar And Red Sindhi) By Microsatellites

by Amber Azam | Miss Sehrish Firyal | Prof. Dr.Masroor Elahi Babar.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Pakistan has vast population of cattle belonging to different breeds. No study on DNA typing of cattle has been conducted in Pakistan. DNA typing of cattle is necessary to overcome the problems like forensic cases, sale & purchase, individual identity in case of fertilization by more than one male and ownership disputes. Microsatellite markers have been proved as an efficient and powerful tool for breed characterization of cattle. In this study, a panel of microsatellite markers, having high polymorphism information content (PlC) values, was developed. Blood samples were taken from jagular vein of two breeds of cattle (Tharparkar and Red Sindhi). DNA was extracted by Inorganic method. Primers of labeled microsatellite markers were optimized for successful amplification conditions in the Bio-Rad thermocycler. Multiplex PCR was performed, for amplification of these labeled microsatellite markers on 44 samples of cattle. (ienotyping analysis was performed for the PCR products of labeled microsatellite markers on agarose gel and then by the genotyper. These results were analyzed statistically software "POPGENE 3.3 and POWER STAT". Allele frequency, heterozygosity, homozygosity and polymorphism information content (PlC) of all microsatellite markers were calculated. Average hetrozygosity, average observed homozygosity and average polymorphism infonnation content (PlC) value for all alleles was 0.60, 0.40 and 0.91 respectively. Almost all of the microsatellite markers showed significant variations in both Tharparkar and Red Sindhi breeds. Microsatellite "1NRAOO5" showed maximum variation i.e. 19 alleles and microsatellite"INRAO23" showed the least variation among all microsatellite markers i.e. 2 alleles. Genotyping results of microsatellite markers were clearly different for two different breeds showing a distinct genetic distance between Tharparkar and Red Sindhi breeds. Results of this study lead to development of a panel of 19 microsatellite markers which can be used for breed characterization of cattle. This was a preliminary study on two cattle breeds (Tharparkar and Red Sindhi) in Pakistan. This facility can be provided on commercial basis to owners. Moreover this study can become the basis for further research investigations on cattle in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 1165,T] (1).

10. Single Nucleotide Polymorphisms &Haplotypic Diversity In The Prkabi Gene Of Pakistani Buffalo Breeds

by Ambrin Fatima | Prof.Dr.Masroor Elahi Babar | Asif Nadeem | Prof.Abu Saeed Hashmi.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2010Dissertation note: Abstract Availability: No items available

11. Bioconversion Of Agriculture Waste To Butyric Acid Through Solid State Fermentation With Clostridium

by Tasleem Akhtar | Dr. Abdu Saeed Hashmi | Dr. Ali Raza | Miss Shagufta Saeed.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Butyric acid is widely used in chemical, food, pharmaceutical industries, in pure form to enhance butter-like notes in food flavors or in the form of esters, as aromatic compounds for the production of perfumes, in dairy products, in the manufacture of cellulose acetate butyrate plastics which is used for textile fiber production, in the treatment of hemoglobinopathies, cancer, and gastrointestinal diseases It is produced in the fermentation by microbial flora living in the large intestine of humans and other monogastric animals. The butyric acid production at industry scale is dominated by chemical synthesis as the starting materials derived from crude oil is currently more attractive due to its low production cost and large scale supply. With the decreasing supply of world crude oil, the increasing supply of food industry by-products which can be used for butyric acid production and the increasing consumer demand for organic natural products in food additives, pharmaceutical products, and preservatives, the production of butyric acid through microbial fermentation has generated again a favorable business. The production of butyric acid was carried out by anaerobic solid state fermentation by C. tyrobutyricum culturing on wheat bran, rice polishing, molasses and corn steep liquor was used as additives.Before its production the proximate analysis of wheat bran, rice polishings, and molasses was carried out to know their inherent nutritional potential. The fermented organism Clostridium tyrobutyricum was isolated from rumen liquor of fistulated buffalo bull .Growth media employed to culture C. tyrobutyricum for the production of butyric acid have been developed. The optimizing conditions of growth medium such ionic concentration of growth medium, source of nitrogen ,substrate to water ratio ,fermentation period and carbon-nitrogen ratio in the medium, for maximum butyric acid production was determined on micro scale at 37°C .Detection and estimation of butyric acid carried out by organic analysis method. This method is based on the catalytic oxidation of butyric acid into diacetic acid, which gives red coloration with sodium nitroprusside. The optimum conditions for the production of butyric acid thus determined on micro scale was applied on higher scale in 7 litre capacity frementer. Availability: Items available for loan: UVAS Library [Call number: 1169,T] (1).

12. Estimation Of Heavy Metals In The Drinking Water Of Residential/Industrial Areas Of Lahore By Atomic Absorption

by Waheed Ahmad | Dr. Abu Saeed Hashmi | Dr. Sualeha | Miss Shagufta Saeed.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2010Dissertation note: Heavy metals are chemical elements with a specific gravity that is at least 5 times the specific gravity of water. The elements studied were mercury, lead, arsenic, cadmium and chromium. Heavy metals have no useful biological function in the body but might be highly toxic as they cause precipitation of proteins especially the enzymes. This investigation was therefore carried out to estimate concentration of these metals and their influence on biological system. For this purpose drinking water samples were collected in one litre polyethylene bottles adding 5 mL of concentrated HNO3 as preservative to adjust the PH<2.00 to maintain heavy metal concentrations during analysis. Samples were marked with unique numbers with dates for the study of Acid Extractable metals. Similarly samples were prepared and preserved for micro biological testing. The metallic ions were estimated by Atomic absorption spectrophotometer of Perking Elmer Model A. Analyst; 2003 at recommended wavelengths for metal ion. Acetylene gas was used as fuel (at 8 psi) and air as an oxidizer. Statistical analysis was done. The calibration curves were prepared separately for all the metals by running suitable concentrations of the standard solutions. It was evident that concentration of chromium, lead, mercury, arsenic and cadmium were high in several drinking water sources in Lahore. This problem is particularly alarming for ground water sources. Almost all water sources are contaminated with lead. According to WHO maximum acceptable limit 10 ppb ,8 water sources had mean chromium concentration in water samples above maximum acceptable limit of WHO (50 ppb), 94 water samples were contaminated with cadmium according to WHO maximum acceptable limit (10 ppb), 13 water sources had arsenic concentration above maximum acceptable limit according to WHO (50 ppb) where as 7 water samples were having concentration of arsenic less than minimum acceptable limit according to WHO (10 ppb) and only 5 water sources meet the criteria of WHO for concentration of mercury, the acceptable limit of 2 ppb. Multitube Fermentation Technique/MPN Method as described by Mackie & McCartney was used for microbiological analysis i.e. Colifcrm bacteria. The results of this study revealed that both samples i.e. tap and ground water do not show conformity with the standards for safe portable water recommended by WHO. The most frequently encountered pathogen in this study was Escherichia Coli which was isolated more in ground water than tap water. It is therefore concluded that by using Atomic Absorption Spectrophotometer concentration of heavy metals in water can be determined and thus on the bases of this work precautionary measures can be taken to prevent the health hazards of these toxic metals. Similarly microbiological analysis of drinking water has provided the evidence that most of the water sources are contaminated with microbes. Availability: Items available for loan: UVAS Library [Call number: 1170,T] (1).

13. Molecular Investigation Of Mental Retardation Locus (Mrti)/Gene Prss12 By Linkage Analysis

by Zafar Ali | Prof.Dr.Masroor Elahi Babar | Dr. Aftab | Mr. Muhammad Asif.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2010Dissertation note: Mental retardation (MR) is a condition in which a person having an intelligence quotient (IQ) lowers than 70. It is also associated with a deficit in adaptive behavior such as communication and daily living skills. Mental retardation is either non-syndromic or syndromic. It is one of the most common genetic disorders and it affects about 1-3% of the human population, with a ratio of males higher than females. The present study was can-ied out to determine the prevalence of families having mental retardation in Pakistani population. In the present study, 7 MR families with three or more affected individuals with MR were enrolled. Family history was taken and pedigree was made personally by visiting the families. The blood samples were collected from the enrolled families. Then DNA was extracted from the blood samples collected from these families by standard inorganic protocol. After isolation of DNA from blood samples, 3 STR markers (D4S191, D4S2392 and D4S3024) in vicinity of mental retardation (MR) locus (MRT1)/gene PRSS12 were amplified on each sample of each family by PCR. The amplified PCR product was first checked on agarose gel and then genotyping analysis (linkage analysis) was performed on non denaturing polyacrylamide gel (PAGE). After polyacrylamide gel electrophoresis, picture of the gel was taken and alleles were read manually with larger allele donated by 2 and smaller by 1. After that haplotype was constructed to determined the pattern of inheritance among the affected and normal individuals of each family under study and also to determined that a family was linked or unlinked to mental retardation locus (MRTI)/gene PRSS12. None of the family was linked to mental retardation locus/gene PRSS12. The families which remain unlinked to the reported loci during screening signifies extreme genetic heterogeneity of MR which is not surprising because about 50% of human protein coding genes are expressed in the brain and it provides an excellent resource material for mapping of the new genes which will shed light on the complex pathways involved in the development of learning and memory in those population. The pedigree of each family in the present study showed that most of the marriages are cousin marriages; therefore this study may play a role in creating awareness about the effect of cousin marriages that is the first step towards decreasing socio-economic burden of the country by genetic counseling and also to prevent mental retardation in Pakistan due to inbreeding. Mental retardation locus (MRT1)/gene PRSS12 was studied for linkage analysis in seven families from different areas of District Swat and Peshawar of Khyber Pakhtunkhwa province of Pakistan. None Out of seven families was linked to mental retardation locus (MRT1 )/gene PRSS 12. All the seven families remain unlinked to this locus. It is concluded that Mental retardation is a complex genetic disorder and needs further studies to identify the already known locus or to explore novel loci through genome wide scan responsible for mental retardation in these population. This will provide opportunities of genetic counseling to these populations and will ultimately result in prevention of mental retardation in Pakistani population. Availability: Items available for loan: UVAS Library [Call number: 1171,T] (1).

14. Identidiation Of Genetic Susceptiblity Of Myopic Loci In Families From Punjab

by Maria Fareed Siddique | Prof.Dr.Masroor Elahi Babar | Dr. Sehrish Firyal | Prof. Dr. Abu.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2010Dissertation note: Myopia, or nearsightedness, is a condition in which the eye cannot focus on distant objects and sometimes closer ones too. In past different authors reported different loci responsible for myopia. They used specifically synthesized markers for different loci and after conducting linkage analysis through genotyping the myopic families were found to be linked for those loci, whereas, in some studies the cause of myopia was environmental. Till now, linkage studies have identified at least 18 possible loci in 15 different chromosomes associated with myopia, although some of these remain to be confirmed. In past, no study was done in Pakistan on myopic families for finding responsible myopic locus in this regard. So, more conclusive and well-designed studies on family pedigrees of individuals with high myopia were needed to be conducted in Pakistan by using genetic markers associated with myopia. In this study, a panel of microsatellite markers was developed. Blood samples were taken from six myopic families. DNA was extracted. PCR was performed for amplification of these I microsatellite markers on 34 samples belonging to 6 families. Genotyping analysis was performed for the PCR products of microsatellite markers. These results were studied by constructing and analyzing haplotypes on the basis of PAGE gel bands. Heterozygosity, homozygosity, polymorphism with all microsatellites markers, specific for two loci were checked. One family MYO-4 was found to be potentially linked with markers for the locus MYP-18. Another family MYO-5 showed potential linkage for the locus 2q37.2. Remaining four families (MYO-l, MYO-2, MYO-3 and MYO-6) were totally unlinked with all the markers (D14S984, D14S63, D14S999, D2S2202, D2S2968 and D2S338 for both loci demonstrating genetic insusceptibility of myopic loci in developing myopia and thus suggesting the complex genetic variability of myopia. This study will serve as the pioneering database for further research on identifying the genetic heterogenic complexity of myopia. Results of this study lead to development of a panel of microsatellite markers which can be used for linkage studies of more myopic families in Pakistan. This study opens the door for new geneticists as the results can also be helpful in carrying out genetic counseling for the myopic persons who are going to be married and specifically for those who have dominant inheritance. This was a preliminary study on myopic patients in Pakistan and data produced during this study will be helpful for drawing and determining genetic inheritance of expected babies with affected parents and siblings. Moreover this study can become the basis for further research investigations on myopics in Pakistan. CONCLUSION This was a pioneering study to develop panel of microsatellite markers for conducting linkage analysis and genetic characterization of myopic patients in Pakistan. As a result of this successful study a reliable, efficient and very informative panel of microsatellite markers was successfully developed which was capable to interpret individual diseased allelic identity, to be used for conducting linkage analysis through genotyping of myopics in Pakistan. This study will serve as the database for further research on identifying the genetic heterogenic complexity of myopia and also these successful results can be further analyzed in future on more myopics from different areas of Pakistan. This work provokes the need for further research purposes in identifying the genes influencing myopia that could help develop targeted treatments for children who are genetically predisposed to developing myopia. Availability: Items available for loan: UVAS Library [Call number: 1177,T] (1).

15. Dna Fingerprinting Of Pakistani Buffalo Breeds (Nili-Ravi, Kundi) Using Microsatellite And Cytochrome B Gene

by Rashid Saif | Prof.Dr.Masroor Elahi Babar | Mr. Asif | Prof. Dr. Abu Saeed Hashmi.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Customarily, classification of breed was based on phenotypic traits. In some cases, recent genetic studies have found differences in the structure proposed. In buffalo ,one has to bear in mind that morphological changes were not the result of adaptation to the environment, but have a social significance thus may not be indicative of the genetic relationship. In recent years microsatellites have proved to be very useful for the determination of genetic relationship among population. Comparative studies beiween microsatellite and protein markers have highlighted the advantages of the former. The water buffalo (Bubalus bubalis) holds tremendous potential in livestock sector in many Asian countries, particularly in Pakistan but the genetic data of different buffalo breeds like Nili-Ravi and Kundi is lacking, which need to be established for their genetic identification. Blood samples of unrelated true representative of both breeds (Nili-Ravi and Kundi) were collected from different government livestock farms in Punjab and Sindh respectively. DNA was extracted by inorganic method and amplification of the mitochondrial Cytb gene and microsatellite was done with especially designed primers in Molecular Biology and Genomics Laboratory at Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore. Cytochrome b gene markers have been proved as an efficient and powerful tool for breed characterization and species identification of buffaloes. Several panels of microsatellite markers have also been reported for this purpose. In this study, a panel of nine microsatellite markers has highly Polymorphism Information Content (PlC) were selected, Specific primers were designed for these microsatellite and Cytb gene partial amplification using primer3 software. Then primers were optimized for successful amplification with minimum reagent concentration. PCRs were performed for amplification of these microsatellite and Cytb markers on each sample, Genotyping and sequencing was conducted on all amplicons to find out the different SNP to design haplotypes with the help of bioinformatics software e.g. Blast 2sequence and Chrornas Lite, Further statistical analysis was done by the help of some other software e.g. Popgene version 1.31, Power Stat., Genetic diversity, Allele frequencies, observed and expected homozygosity and heterozygosity, Hardy Weinberg equilibrium, F-Statistics and Gene Flow for all Loci, population's dendogram, Nei's genetic identity and genetic distance! diversity was calculated. The results obtained from this study can contribute to the establishment of routine DNA typing services, beneficial for the buffalo industry as well as in animal forensics for litigation and expedite the police investigation services in Pakistan, which will also be useful for breed characterization and phylogenetic study of aforementioned breeds of buffalo. Availability: Items available for loan: UVAS Library [Call number: 1183,T] (1).

16. The Study Of Gene Gjb2/Dfnb1 Causing Deafness In Humans By Linkage Analysis From District Peshawar

by Noor Badshah | Prof.Dr.Masroor Elahi Babar | Dr. Aftab | Mr. Muhammad Asif.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2010Dissertation note: Hearing impairment is the partial or complete inability to hear that leads to compromise the development of normal language skills. Among all the sensory impairments in humans, hearing impairment is the most common. It is estimated that at least 50% of the cases are due to genetic factors. Hereditary hearing loss may be syndromic or non-syndromic; about 30% of deafness cases are syndromic, while 70% is non-syndromic. It is estimated that the prevalence of profound bilateral hearing loss is 1.6 per 1000 in Pakistan and 70% of hearing loss arises in consanguineous families. The main pattern of inheritance of deafness in Pakistani population is autosomal recessive and to date more than 145 loci and 26 genes have been identified for non-syndromic recessive deafness. More than 400 disorders associated with hearing loss shows extreme genetic heterogeneity and complexity of the mammalian inner ear. As more genes are identified, the elucidation of the function of the proteins that these genes encode contributes greatly to the understanding of cochlear mechanisms and their role in disease causation. The gene involved, GJB2, encodes the connexin26 molecule. Connexin26 is a component of gap junctions, the links that allow small molecules to pass from one cell to the next, and this protein is found in several places in the body, including the epithelial supporting cells surrounding the sensory ear cells of the cochlea.The sensory ear cells of the cochlea allow potassium ions to pass through their upper surface during normal reception of sound, and these potassium ions must be recycled through the base of the ear cells and the supporting cells and fibrocytes back into the high-potassium endolymph that bathes the tops of the ear cells. The aim of this study was Linkage analysis for DFNB1 locus involved in causing hereditary deafness in families from Khyber Pukhtunkhwa. A total of 10 families were enrolled from different areas of Khyber Pukhtunkhwa province. I have studied 8 families of these 10 (i.e.) family no. 2, 3, 4, 5, 6, 8, 9 and 10. The families have at least three affected individuals. All the families showed recessive mode of inheritance. For linkage analysis studies for DFNB1 locus, three STR markers D13S175, D13S292, and D13S787 were genotyped using Polyacrylamide gel electrophoresis (PAGE) and haplotypes were constructed to determined, linkage with DFNB1 locus. From a total of 8 families, a single family was linked to DFNB1 locus. The DFNB1 locus is the first non-syndromic deafness locus mapped to chromosome 13q12. Availability: Items available for loan: UVAS Library [Call number: 1191,T] (1).

17. Molecular Diversity Analysis Of Sheep And Goat Breeds Of Pakistan Using Microsatellites.

by Misbah Shaheen | Prof.Dr.Masroor Elahi Babar | Mr. Tanveer Hussain | Prof. Dr. Azhar.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Pakistan is rich in Animal Genetics Resource (AnGR) and has various breeds of sheep and goat but the genetic data in these different breeds is lacking which needs to be established for their genetic identification. The advent of molecular techniques has led to an increase in the studies that focus on the genetic characterization of domestic breeds using genetic markers. Due to their reliability and availability, the microsatellites have become preferred method for the genome mapping. Microsatellites or STRs are the 1-6 nucleotide tandem repeats present in both coding and non coding regions of both prokaryotes and eukaryotes. Microsatellites are powerful tools in genome mapping, forensic DNA studies, paternity testing, population genetics and conservation! management of biological resources. The present study was conducted on the molecular diversity analysis of sheep and goat breeds of Pakistan using FAQ recommended unlabelled microsatellites. Blood samples of unrelated true representative animals of two sheep and goat breeds were selected from their breeding tracts and different Government Livestock Farms throughout the country. DNA was extracted with the standard protocol and amplification of DNA was done with a set of 16 microsatellite markers in Molecular Cytogenetics and Genomics Laboratory in the Institute of Biochemistry and Biotechnology. The products of touch-down PCR were examined on non denaturing Polyacrylamide Gel Electrophoresis (PAGE). Genotyping results were analyzed through the sofiware POPGENE version 3.3 for calculating the number of alleles, expected and observed heterozygosity, homozygosity, Polymorphic Information Content (PlC). Average observed heterozygosity, average observed homozygosity, observed and effective number of alleles for all loci and populations were 0.8394, 0.1606, 3.6875 and 2.8693 respectively. Almost all of the microsatellite markers showed significant variations in both breeds of sheep and goat. Genotyping results of microsatellite markers were clearly different for four different breeds showing a distinct genetic distance between sheep and goat breed's. This work provided the genetic data which will be helpful in breed identification and making effective breeding policies and conservational activities in future according to FAO global Farm Anithal Genetic resource data. Moreover this study can become the basis for further research investigations in sheep and goat in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 1200,T] (1).

18. Production, Purification And Concentration Of Rabbit Anti Goat I&G Antibodies

by Yasir Ashrif | Dr. Abdu Saeed Hashmi | Dr. Ali Raza | Ms. Faiza Masood.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Antibodies are not only important in medical research but these are also important in treatment. In this study, the production, purification and concentration of polyclonal immunoglobulin G (IgG) antibodies against goat IgG immunoglobulins in rabbits was carried out. Partial purification of goat IgG obtained at 33 % ammonium sulphate saturation was 2.43 mg/mL. It was followed by Diethylaminoethyl (DEAE) cellulose ion exchange chromatography which gave purified fraction of IgG. Then it was concentrated by polyethylene glycol (PEG) and now the IgG concentration was found 3.17 mg/mL. After purification, different doses of IgG in combination with adjuvant were injected into nine, 8 months old rabbits. After immunization of rabbits, the blood samples were collected and antigoat IgG was purified as described above, this rabbit antigoat IgG concentration after purification was found 3.26 mg/mL. Production of these anti-IgG antibodies were tested by agar gel precipitation test (AGPT) and radial immunoassay. The titer of AGPT with goat and rabbit serum was 256 and the titer with IgG was 32. The purity of IgG was confirmed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), by obtaining 50 KDa bands of IgG heavy chains and 25 KDa bands of light chains. However, this purified rabbit anti-goat IgG when conjugated with horse radish peroxidase can serve to diagnose various microbial infections of goat through ELISA. Availability: Items available for loan: UVAS Library [Call number: 1197,T] (1).

19. Dominant Inheritance Of Myopia Linked Loci Myp10 And Myp11

by Saira Nabi | Mr. Asif Nadeem | Mr. Muhammad Imran | Prof.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Myopia, a refractive error is one of the most common ocular disorders worldwide with elongation of axis of the eyeball. Genetic plays an important role in the development of myopia. Familial myopia is common in Pakistan. The aim of the study was to find out the dominant inheritance through linkage analysis and molecular characterization of Myopia in Pakistani myopic families and also to determine the presence of loci (MYP 10 and MYP1 1). Six families with three or more affected individuals in two or more ioops were enrolled from different areas of Lahore. The persons with refractive error equal or worse than -l were considered as myopic. Out of the total 36 samples, 26 were myopic. For the each locus, 3 markers were designed .Polyacrylamide Gel electrophoresis (PAGE) was used for genotyping of amplified DNA samples. Haplotypes of all the families were constructed based on the PAGE results to check weather a family is linked or unlinked to those loci. Result of allele haplotyping were analysed to evaluate the linkage of Myopia loci and dominant inheritance in Pakistani families. No linkage was found for any of those selected families. The negative results for these chromosomal regions have several possible reasons; sample size, family size and ethnicity of the families are the major reasons for these negative results. Although there was no linkage for the loci MYP10 and MYP11, this would be the first molecular investigation of the Myopia loci MYP10 and MYP11 in Pakistani families. The findings of the proposed study will be vital for victim familes in terms of genetic testing, genetic counseling, for designing a management plan and resource alloction for victims. Availability: Items available for loan: UVAS Library [Call number: 1215,T] (1).

20. Linkage Analysis Of Myp12 And Myp14 In Families From Lahore

by Maryam Zahra | Prof.Dr.Masroor Elahi Babar | Dr. Ali Raza Awan | Prof. Dr.

Material type: book Book; Format: print 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).

21. Genetic Study Of Ushic/Dfnb18 By Linkage Analysis

by Muneer Ahmad | Mr. Tanveer Hussain | Prof. Dr. Masroor Elahi Babar.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2010Dissertation note: Deafness refers to conditions in which individuals are fully or partially unable to detect or perceive at least some frequencies of sound which can typically be heard by members of their species. In human beings, the term hearing impairment is usually reserved for people who have relative insensitivity to sound in the speech frequencies. In the present study, six families were identified which were collected from FATA, Lahore and Sheikhupura and were consist of at least two deaf people. Pedigrees of the affected families were drawn using Cyrillic software. Blood samples were collected from these families. DNA was extracted through Inorganic protocol. The creening of the affected families was done for known deafness locus, USH1C/ DFNBI8. Then the PAGE (Polyacrylamide Gel Eletrophoresis) was done and haplotypes were constructed to determine whether a family was linked to deafness locus or not. Out of six families, no family was linked to USHIC/ DFNB18 locus. Availability: Items available for loan: UVAS Library [Call number: 1223,T] (1).

22. To Study Influence Of Butyric Acid On The Performance Of Broiler Chicks

by Asiya Akbar | Dr.Abu Saeed Hashmi | Miss Shagufta Saeed | Prof.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Poultry have been on the earth for over 150 million years, dating back to the original wild jungle fowl. Poultry provide humans with companionship, food and fiber in the form of eggs, meat and feathers. There is a large commercial chicken industry that provides us with eggs and meat. Because poultry products are in demand around the world and because chickens and other poultry can be reared in almost any part of the world, a renewed interest in poultry projects for schools, 4-H groups and in the home has developed. The butyric acid is an excellent growth promoter as it is an efficient nutrient for the intestinal mucosa increasing the density and length of villi, enlarging the adsorption surface of the intestine. It is also known as antibacterial agent against pathogenic microorganisms including Salmonella, Escherichia coli, Brachyspira etc. and as modulator of the intestinal flora supporting useful microorganisms such as lactobacilli. Rice polishings was used as a substrate for the production of butyric acid and corn steep liquor as additive. Solid state anaerobic fermentation process was used for butyric acid production through C. tyrobutyricum according to the predetermined optimized conditions. Estimation of butyric acid was carried out by organic analysis method by Deniges (1918).This method is based on the catalytic oxidation of butyric acid into diacetic acid ,which gives red coloration with sodium nitroprusside. Effect of butyric acid on the performance of Broiler chicks was studied by conducting feeding trial on 132 day old broiler chicks purchased from commercial hatchery and was randomly subdivided into 12 units, 11 chicks each. Four diets "A", "B", "C" and "D" (table) were constructed whose composition is as follow. "A" diet was served as control, while the "B", "C" and "D" diets were supplemented with 0.2%, 0.4% and 0.6 % butyric acid respectively. These diets were randomly assigned to the above chicks for a period of 42 days as feeding trial. The performance of birds was monitored in terms of weight gain, feed efficiency, protein efficiency and dressing percentage. From the results obtained it is concluded that O.4 % butyric acid gave maximum gain in weight, feed efficiency ratio and dressing percentage. Hence it can be stated profoundly that the ration containing 0.4 % butyric acid has stimulated the gastric secretions which ultimately had improved the performance of broiler chicks. Availability: Items available for loan: UVAS Library [Call number: 1251,T] (1).

23. DNA Typing And Quantification From Different Saliva Stained Substrates In Central Punjab

by Mudassar Naseer | Dr.Atif Hanif | Dr.Aftab | Prof.Dr.Masroor Elahi Babar.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Biological evidences have most significant place in forensic analysis. These biological evidences may be blood, hair, tissue, nails, and saliva. Pakistan is one of the highest populated country having poor Law & Order Situation. DNA typing is the best tool for human identification all over the world which is unfortunately is at scant in Pakistan at the Moment. Biological evidences have most significant place in forensic analysis. These biological evidences may be blood, hair, tissue, nails, and saliva. In present study the extraction, quantification and typing of DNA was done from different fruit pits and peels from forensic point of view. DNA as biological evidence have been collected from cigarette buts, bottle necks, bottle straws, tea cups but not from any fruits pits or fruit peels so it will be a new study in forensic field. DNA extraction from these saliva stained samples was done by organic method. Quantification was done by Real Time PCR using Quantifiler Kits. Identifiler Kit having 15 STRs was used for Genotyping of amplified products. Statistical analysis done using POP GENE and Power Stats softwares. Allele frequency, heterozygosity, homozygosity, polymorphism information content (PIC), power of discrimination and power of exclusion of all microsatellite markers were calculated. Average power of discrimination, average hetrozygosity, average observed homozygosity and average polymorphism information content (PIC) value for all alleles 0.856, 0.7158, 0.2238, 0.71, respectively. Moreover combined power of exclusion reached a significant value of 0.9998. Almost all of the microsatellite markers showed significant variations in population size Microsatellite "D8S1179, D2S1338, and FGA" showed maximum variation i.e. 8 alleles and microsatellite"D18S51 and THO1" showed the least variation among all microsatellite markers i.e. 4 alleles. Genotyping results of microsatellite markers were clearly different for each donor of the saliva stains. This study was done on preliminary basis in Pakistan and successful results were obtained for forensic analysis of the any crime scene. This facility can be provided on commercial basis as forensic tool and the research based study can be extended to a number of stains of the biological secretions in the same directions. Availability: Items available for loan: UVAS Library [Call number: 1253,T] (1).

24. Age And Gender Related Response To Antiviral Therapy Against Hcv Genotype 3A

by Saba Manzoor | Prof.Dr.Masroor Elahi Babar | Ms Aseeda | Prof.Dr Muneer Saleemi.

Material type: book Book; Format: print Publisher: 2010Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1271,T] (1).

25. Decolorization Of Effluents From Textile Dyes By Fungi

by Ghulam Rasool Anjum | Dr.Abu Saeed Hashmi | Dr.Ali Raza | Mr.Zahid Mushtaq.

Material type: book Book; Format: print ; Nature of contents: biography Dissertation note: Textile sector is one of the large exporting and the most important industrial sector of Pakistan. It accounts about 64% of the total export from our country. At present there are 650 textile processing units working in Pakistan. Textile industries consume large amount of water for wet processing of textile. Out of many contaminants present in wastewater, such as acids, bases, toxic organic and inorganic dissolved solids, and colors, Colors are considered the most undesirable and are mainly caused by dyes which are used in textile dying industry. The objective of the study was to decolorize the cotton industry waste water effluents by treatment with microorganisms like Trichoderma harzianum and Aspergillus flavus, an extent to make it least harmful to water habitats and also to make it fit for irrigation purposes. The influencing parameters that affect the percentage of decolorization rates are optimized in still culture fermentation. Spectrophotometric analysis method was used to estimate decolorization of textile industry effluent at its ?max (350nm). The optimal values of the parameters such as effluent to water ratio, fermentation time, pH, and carbon to nitrogen ratio are found to be 1:0, 72 hours, 4.0, and 1: 2.33 for Trichoderma harzianum and 1:1, 72 hours, 5.0, and 1: 2.33 for Aspergillus flavus respectively. The concentration of different ions like Ca+2, Mg+2 and H2PO4-1 were also optimized for maximum decolorization and the optimized conc. were 0.025%, 0.025% and 0.025% for Trichoderma harzianum and 0.05%, 0.1% and 0.05% for Aspergillus flavus respectively. The maximum percentage of decolorization at the optimized conditions on large scale was found to be 83.31% and 80.02 % with Trichoderma harzianum and Aspergillus flavus respectively. Availability: Items available for loan: UVAS Library [Call number: 1279,T] (1).

26. Decolrization Of Textile Dyeing Industries Effluent By Ganoderma Lucidum And Arachniotus

by Sobia Majeed.

Material type: book Book; Format: print Publisher: 2011Dissertation note: The textile industry is a water intensive industry with water being used in every stage of wet processing from sizing, designing, scouring and bleaching of fibers to the dyeing, finishing and printing of fabrics. The objective of this study was to decolorize the cotton industry waste water effluents by treatment with microorganisms like Ganoderma lucidum and Arachniotus sp. to an extent to make it least harmful to water habitats and also to make it fit for irrigation purposes. The influencing parameters that affect the percentage of decolorization rates are optimized in still culture fermentation. Spectrophotometric analysis method was used to estimate decolorization of textile industry effluent at its ?max (410 nm). The optimal values of the parameters such as effluent to water ratio, fermentation time, pH, and carbon to nitrogen ratio are found to be 1:0, 72 hours, 6.0, and 1: 2.33 respectively. The concentration of different ions like Ca+2, Mg+2 and H2PO4-1 were also optimized for maximum decolorization and the optimized conc. were 0.05%, 0.05% and 0.1% for Arachniotus sp. and 0.1%, 0.05% and 0.1% for Ganoderma lucidum respectively. The maximum percentage of decolorization at the optimized conditions on large scale was found to be 82.24% and 81.32 % with Arachniotus specie and Ganoderma lucidum respectively. Availability: Items available for loan: UVAS Library [Call number: 1282,T] (1).

27. Characterization Of Dgati Gene For K232A Polymorphism In Pakistani Cattle Breeds

by Rashid Hussain | Prof.Dr.Masroor Elahi Babar | Dr.Aftab Anjum | Mrs.Saeeda Kalsoom.

Material type: book Book; Format: print Publisher: 2011Dissertation note: 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. DGAT1 gene has important role in milk fat percentage. This gene is located on the centromeric end of the bovine chromosome 14. It was recently studied that nonconservative dinucleotide (AA?GC) substitution in exon 8 of DGAT1 gene, change lysine to alanine at position 232 (K232A mutation) of the encoding protein. Animals that have K (Lysine) at position 232 in amino acid sequence of DGAT 1 will have high fat percentage and low milk yield as compared to animals that have A (Alanine) at this position. The objectives of this study were to identify K232A (Lysine232?Alanine) polymorphism in DGAT1 gene in Pakistani cattle breeds. To find relationship between milk production and K232A polymorphism. Blood samples were collected from different Govt. livestock farms/experimental stations. DNA was extracted by organic method. Specific primers were used for the amplification of exon8 of DGAT1 gene. After DNA amplification by Polymerase Chain Reaction, restriction digestion was done by using CfrI enzyme. Fourty animals belonging to Sahiwal and Dhanni breeds were genotyped for K232A polymorphism by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) technique. Sequencing was carried out to confirm the results of restriction digestion. From the data analyzed it was observed that all the animals of Sahiwal and Dhanni breeds had K allele and no A allele was identified. It has been shown that a missense mutation (Lys232 ? Ala) in the bovine DGAT1 gene in Pakistan cattle breeds is not common. Availability: Items available for loan: UVAS Library [Call number: 1283,T] (1).

28. Production Of Methane Gas From Effluent Of A.B Murie By Methanobacterium Ruminantium

by Huma Shabbir | Dr.Abu Saeed Hashmi | Miss Saeeda | Miss Shagufta Saeed.

Material type: book Book; Format: print Publisher: 2011Dissertation note: There is extreme shortage of energy in our country. Homes, private sectors and industries are suffering a lot due to this shortage. There are different sources of energy like fossil fuels which include coal, petroleum and natural gas etc. Apart from it there are number of renewable sources but fact is that all these sources are getting diminished because of excessive use due to growing population of the world. In order to increase energy in our country it is imperative to utilize the industrial waste. At present a lot of industrial effluent is being wasted which can be used for the production of fuel in the form of methane gas. The production of methane was carried out by anaerobic fermentation by Methanobacterium ruminantium culturing on Effluent of A.B Murie. Before its production the proximate analysis of effluent was carried out to know its inherent nutritional potential. The fermented organism Methanobacterium ruminantium was isolated from feces of buffalo bull. The optimizing conditions of growth medium such as temperature, pH, ionic concentration of growth medium (Ca+2, Mg+2) and Carbon-Nitrogen ratio in the medium, for maximum methane production was determined on micro scale. Detection and estimation of methane was carried out by burning and by measuring pressure on pressure gauge. The optimum conditions thus determined on micro scale was applied on higher scale in a 3 liter flask. The pressure of methane gas obtained was 1.3 psi. It is therefore concluded that effluent of A.B Murie contain sufficient potential of methane production. The methane thus produced can be used as a fuel and light source for their own use. Availability: Items available for loan: UVAS Library [Call number: 1285,T] (1).

29. Genetic Diversity Analysis Of Sahiwal And Dhanni Cattle Breeds By Cytochrome B Gene And Microsatellite Markers

by Zahoor Ahmed | Prof.Dr.Masroor Elahi Babar | Mr. Tanveer Hussain | Prof.Dr.Muham.

Material type: book Book; Format: print ; Nature of contents: biography; Literary form: Publisher: 2010Dissertation note: Pakistan has various dairy breeds of cattle but the genetic data of different cattle breeds including Sahiwal and Dhanni is lacking which need to be established for their genetic identification. Blood samples of unrelated true representative of breeds (Sahiwal and Dhanni) were collected from their respective home tracts and different Government livestock farms. DNA extracted with the standard protocol (Inorganic Method) in Molecular Biology and Genomic Laboratory, Institute of Biochemistry and Biotechnology (IBBT), University of Veterinary and Animal Sciences, Lahore. Nine fluorescent dye labeled microsatellite markers having high polymorphism information content (PIC) values were used and genotyping was done. These results were analyzed statistically by softwares "POPGENE 1.31 and POWER STAT" 2.1. Allele frequency, heterozygosity, homozygosity, polymorphism information content (PIC), power of discrimination, power of exclusion, F-Statistics and Gene Flow for all Loci, population's dendogram, Nei's genetic identity and genetic distance/ diversity were calculated. The average observed heterozygosity was 0.5845 and 0.5911 in Dhanni and Sahiwal respectively. The mean observed homozygosity was 0.4155 and 0.4089 in Dhanni and Sahiwal respectively. The average PIC (Polymorphic Information Content) values of nine loci showed by Dhanni and Sahiwal cattle are 0.61 and 0.77 respectively. Genotyping results of microsatellite markers were clearly different for two different breeds showing a distinct genetic distance between Dhanni and Sahiwal cattle breeds. For further confirmation of the breeds amplification of the mitochondrial Cyt b gene was done with especially designed primers which were designed by using Primer3 software. Sequencing of PCR fragments was done. Analysis of the sequences was performed by multiple sequence alignment with the help of Blast 2sequence and BioEdit soft wares. Identified SNPs were analyzed and haplotypes were formed. Phylogenetic tree was constructed by MEGA 4.1. The use of genetic markers provided the information on population genetic structures of the indigenous cattle breeds even if they lack detailed pedigree recording data. The study on the genetic diversity showed the differentiation of breeds and individual breeds have unique combinations of genes as a result of phylogenetic tree. This work will provide the genetic data which will be helpful in breed identification and making effective breeding policies and conservational activities in Pakistan in future according to FAO global Farm Animal Genetic resource data. Availability: Items available for loan: UVAS Library [Call number: 1289,T] (1).

30. Detoxification Potential Of Yeast Sludhe Ahainst Ochratoxin In Broiler Chicks

by Huma Mujahid | Dr. Abu Saeed Hashmi | Dr. Aftab | Miss Asma Waris.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Ochratoxin the fungal secondary metabolite is a potent natural contaminant of poultry feed. Mycotoxins present in poultry feeds from the raw material used in their production is the major cause of toxic feed. The intake of very low levels of Ochratoxin-A result in overt ochratoxicosis resulting in impairment of immune system and acquired resistance to infections causing health problems which lead to economic losses in the form of reduced productivity The research study was conducted to study the harmful effects of Ochratoxin on broiler chicks and the adsorptive potential of yeast sludge against Ochratoxin in broiler chicks . Aspergillus ochraceus was grown on Sabraud's Dextrose Agar and ochratoxin was produced on fermented wheat grains .One fifty day old Chicks of broiler breed were purchased from Big birds hatchery and were raised on commercial broiler diet till 7 days. Four diets A,B,C and D were formulated A diet serve as control, B diet contained OTA 500ppb, C diet contained OTA 500ppb and 1% Yeast sludge and D diet contained OTA 500ppb and 2% Yeast sludge. These four diets were assigned randomly to the chicks, such that there were three replicates on each ration and each replicate contained 10 chicks. Vaccination against N.D and IBD was performed according to the schedule. During feeding trial weight gain , feed consumed, FCR and mortality rate was determined. Group B (500ppb OTA) showed a decrease in weight gain and feed consumption as compared to group A (control diet) , C (1% yeast sludge and 500ppb OTA) and D (2% yeast sludge and 500ppb ochratoxin). Group D showed more improvement in weight gain, feed consumption and FCR as compared to group C. Blood serum and tissue samples were collected from the birds slaughtered at the end of experimental trial. Concentration of serum total protein, albumin and activity of alanine transaminase were determined. Blood Serum levels of total protein and albumin were lower in the group B (500ppb OTA) than group D having 2 % yeast sludge but the group C fed on 1% yeast sludge did not show much improvement in those parameters. Activity of ALT was found to be significantly higher (P<0.05) in group C as compared to all other groups. Whereas blood serum ALT activity of the birds fed on ration B was significantly high (P< 0.05) as compared to blood serum ALT of group A The Level of Ochratoxin in Liver and Kidneys was also determined and it was found to be highest in Group B (500ppb OTA) and lowest in Group D (500ppb OTA + 2% yeast sludge). Based upon the observations obtained in this study it can be concluded that ochratoxin-A is a nephrotoxic and hepatotoxic agent. But supplementation of 2% yeast sludge in the broiler diet can effectively detoxify the effects of ochratoxin as compared to supplementation of 1% yeast sludge in the chicks diet. Availability: Items available for loan: UVAS Library [Call number: 1313,T] (1).

31. Antibacterial Activity Of Indihenous Hernal Exteacts Ahainst Urease Profucinh Bacreria

by Rubina Yasmeen | Dr. Abu Saeed Hashmi | Dr. Aftab | Miss Shagufra Saeed.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Poultry farming is a profitable business but is facing serious ammonia environment particularly during winter season when ventilation frequency is reduced to maintain the shed's temperature. Urease producing bacteria in droppings are main cause of emitting ammonia in the sheds. The ammonia poultry environment is inducing reduced weight gain, immuno-suppression, enhanced susceptibility to respiratory pathogens, etc. Aqueous and alcoholic extracts of 14 local herbs (Aloe Vera, Azadirachta indica, Allium sativum, Calotropis procera, Cannabis sativa, Carum capticum, Eucalyptus camaldulensis, Lantana camara, Mangifera indica, Mentha piperita, Nigella sativa, Opuntia ficus indica, Piper nigrum and Zingiber officinalis) and four commercial herbal products (Mentofin, Suduri, Safi, Yucca) were evaluated for their in-vitro antibacterial activity against Proteus mirabilis by serial dilution method. It was observed that with reference to rise in pH, Ammonia concentration and urease activity in aqueous and alcoholic extracts of Allium sativum (pH: 8.5560, 8.8480, Ammonia:4.42, 3.52 µg/mL, Urease: 0.009, 0.007 U/mL respectively) had shown best results as compared to control positive (pH: 9.03, Ammonia: 6.7µg/mL, Urease: 0.013 U/mL). Alcoholic extracts of Mangifera indica (8.8820, 5.42µg/mL, 0.010 IU/mL), Mentha piperita (8.8880, 4µg/mL, 0.008 U/mL) Carum capticum (8.9540, 4.84µg/mL, 0.009 U/mL) and aqueous extract of Opuntia ficus indica (8.8100, 5.22µg/mL, 0.010 U/mL) had weak activity against P. mirabilis. Both aqueous and alcoholic extracts of Eucalyptus camaldulensis (pH: 8.91, 8.96, Ammonia: 5.16, 5.06 µg/mL, Urease: 0.01, 0.01 U/mL) has weak inhibitory effect. All commercial products had shown strong antibacterial activity (pH: 4.8-6.8, Ammonia: 0µg/mL, Urease: 0 U/mL). Results of remaining herbal extracts were not significantly different (p<0.05) from positive control. It was concluded that all herbal products had strong antibacterial activity against P. mirabilis. Mentofin had shown best results with optimum inhibitory concentration (1/1000 mL). Alcoholic extracts of few herbs had shown weak bactericidal activity. These herbs might give better results in-vivo. Availability: Items available for loan: UVAS Library [Call number: 1314,T] (1).

32. Isolation And Characterization Of Collagen Type Ii From Poultry Trachea

by Sidra Ashraf | Dr. Abu Saeed Hashmi | Dr. Sualeha Riffat | Zahid Mushtaq.

Material type: book Book; Format: print Publisher: 2011Dissertation note: This project was designed to use poultry waste to isolate and characterize collagen type II from its trachea. Collagen type II is being used along with condroitin sulfate and glucosamine for the treatment of osteoarthritis and is also available as a neutraceutical product in the market. For project purpose, trachea of slaughtered broiler birds were collected from the market and after removing adhering tissue and debris, it was then washed thoroughly first with distilled water and then with deionized water. Tracheal cartilage was then cut into small pieces and defattened with chloroform: methanol (2: 1 v/v) solution. After this, the cut pieces were properly cleaned with deionized water. 0.5% Pepsin solution in 0.5 M acetic acid was prepared. Cartilage was then hydrolyzed by the already prepared 0.5 % pepsin (in 0.5 M acetic acid) at 4 ° C for 48 hours. The extract was then separated from the tracheal pieces and the viscous solution obtained was centrifuged at 12000 rpm for 1 hr at 4 "c. Now the collagen was expected to be in the supernatant which was salted out by adding NaCI to a final concentration of 2.5M and kept for almost 12-16 hrs. This collagen was again centrifuged at 12000 rpm for 1 hr at 4 C. The obtained collagen pallet was redissolved in 0.5 M acetic acid and then it was dialyzed against 0.1 M acetic acid followed by dialysis with distilled water. The sample after dialysis was put in petri dishes and kept in freezer for overnight to let it be prepared for lyophilization. The frozen collagen sample was then lyophilized. After lyophilization, the sample gave an appearance of a white mesh. This sample was reconstituted in PBS with pH 8 to run it on SDS-PAGE. The procedure of SDS-PAGE in non reducing conditions was adopted for the characterization of collagen type II in the sample. The description of results of SDS-PAGE is given below: Lane M contains protein markers of different molecular weight. Lane 1, 2 and 3 contains samples at different steps of the whole procedure showing clear bands of collagen type II. Lane 4 contains lyophilized sample of collagen type II showing the thickest band (alpha chain of collagen type II). In this research, poultry waste has been used for making health improving product. As in our country poultry is used in bulk quantity so if its waste might be used in any medicinal product then it might not only be useful but also economical for such a developing country as ours. Another thing is that as this collagen Type II has been extracted from poultry trachea, it shows that tracheal cartilage is a rich source of such collagen type. Collagen Type II is used in the cure of arthritis especially rheumatoid arthritis so through this research, it has been made clear that poultry waste can be utilized in a positive way in medicinal industry and also that collagen Type II acts as an effective neutraceutical. Availability: Items available for loan: UVAS Library [Call number: 1330,T] (1).

33. A Study On Prevalance Of Hcv Genotypes And Risk Factors Of Hepatitis C Virus In Punjab

by Tahira Tarar | Miss Faiza Masood | Dr. Muhammad | Mr. Zahid Mushtaq.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Hepatitis C virus (HCV) was marked as major agent which causes non-A, non-B hepatitis. Various partial and complete sequences of HCV nucleotide had been identified in the world. When these sequences were compared a marked genetic heterogeneity was revealed, that suggested the existence of HCV genotypes. Recent studies have pointed out the association of different HCV genotypes with different profiles of pathogenecity, infectivity and response to antiviral therapy. In our study we used typing system based on genotype specific primers focused on HCV 5´-UTR by using PCR. Genotype specific primers were designed for genotype 1, 2a, 2b, 3a, 3b and 4. 100 samples of HCV positive patients were collected. The frequency of occurrence of genotype 2a was 4 %, 2b was 5 %, 3a was 71 %, 3b was 11% and untypeable was 9 %. Blood transfusions, therapeutic injections, reuse of needles, dental procedures, shaving practices, body piercings, jaundice, dialysis, surgery and other multiple risk factors associated with HCV were studied. Major risk factor among females was therapeutic injections and among males the major risk factor was absence of shaving precautions. The study concludes that the most prevalent genotype in Punjab province is 3a. As each genotype sequence is different, the antiviral therapy against that particular genotype is also different. The treatment would only be successful if the genotype of HCV infected patient is known. This study will help in correlating efficacy of interferon therapy with different HCV genotypes and to understand the mode of transmission for hepatitis C virus. Availability: Items available for loan: UVAS Library [Call number: 1336,T] (1).

34. Dna Typing And Quantification Of Lip Cosmetic Samples For Forensic Casework Analysis

by Rabia Umer | Mis. Saeeda Kalsoom | Prof. Dr.Masroor Elahi Babar.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Biological evidences have most significant place in forensic analysis. These biological evidences may consist of blood, hair, tissue, nails and saliva. While lip cosmetics can also become important non-biological source in certain cases. Considering this fact, present study was designed to get DNA from different sources of lipstick and its impressions. DNA as biological evidence had been collected from lips after using lipstick, used lipstick, eaten-apple, bite-mark from arm, facemask and tissue with lipstick impressions, cigarette butt, pen, kiss and glass from female donors. Buccal swabs were also taken as reference. DNA extraction from these lipstick stained samples was done by organic method while quantification was performed through RT-PCR by using Quantifiler assay. All amplifications were performed using (IdentifilerTM Kit having 16 STRs) GeneAmp PCR System 9700 thermal cycler for 28 cycles and the amplified product was analyzed with ABI Prism 3100 Genetic Analyzer and GeneMapper® ID analysis software v3.2. All profiles were successfully matched with their reference DNA. Present study may help to develop a tool for extraction and quantification of DNA from trace evidence like lipstick obtained from crime scenes in forensic analysis. This study was done on preliminary basis in Pakistan and successful results were obtained for forensic analysis. Furthermore, this research may be extended by increasing number of samples, gender discrimination and detection of SNP in samples generating partial profile. Availability: Items available for loan: UVAS Library [Call number: 1338,T] (1).

35. Genetic Study Of Myp 2, Myp 15, Myp 16, And Myp 17, Loci Of Myopia In Families Of Province Punjab

by Uzma Naureen | Ms. Saeeda Kalsoom | Prof. Dr.Masroor Elahi Babar.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2011Dissertation note: Myopia is said as the common cause of impaired vision and visual disability. In this disease the image is not focused sharply on the retina causing a dim 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 [our billion in Pakistan. It is multi factorial disease and 19 loci identified up to date. Eight myopic families were identified and selected for this study from different areas of Punjabprovince. Linkage analysis of these families was done by MYP 2, MYP J 5, MYP J 6 and MYP J 7 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 12 microsatellite markers were used to analyze 40 samples from eight families. After DNA extraction and peR amplification, linkage analysis was carried out by gcnotyping through PAGE and haplotypes were constructed for the fami lies. Availability: Items available for loan: UVAS Library [Call number: 1347,T] (1).

36. Production Of Alginate By Azotobacter Vinelandll By Submerged Fermentation

by Muhammad Awais | Miss Shagufta Saeed | Miss Asma Waris | Miss Sehrish.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Alginate is a linear coploymer of 1, 4,-beta-D-mannuronic acid and alpha -L-guluronic acid. They are usually obtained from brown algae which contains of D-mannuronic and L-guluronic acids. Alginates are important polysaccharides with many application in the food, pharmaceutical, textiles and paper industry. Azotobacter vinelandii is a gram negative, aerobic and free-living soil microbe which play an important role in the nitrogen cycle in nature. A. vinelandii was grown on wheat bran for the production of alginate. The optimal condition like substrate:water ratio, pH, incubation period, ionic concentration (Calcium, magnesium, sodium and phosphate ions) and addition of corn steep liquor was carried out for the production of alginate by A. vinelandii through submerged fermentation. The cell biomass and alginate concentration was determined to estimate the concentration of alginate. The alginate thus produced was estimated by determining its concentration spectrophotometrically at 546 nm. The effect of pH, incubation period, ionic concentration and addition of nitrogen source like corn steep liquor was analysed by statistical linear regression method. Alginate is an important polymer with wide applications in pharmaceutical, food and textile industry. It has also medical application in encapsulating hormones. Availability: Items available for loan: UVAS Library [Call number: 1348,T] (1).

37. Genome-Wide Association Mapping To Approach The Candidate Gene Having Potential Role In Dairy Bull Fertility

by Asif Nadeem | Prof.Dr.Masroor Elahi Babar | Dr.Atif Hanif | Prof.Dr.Muhammad Abdullah.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Reproductive efficiency is a most important determinant of dairy profitability. Fertility in the herd is absolutely critical for both male and female animals. Fertility studies in dairy cattle were directed toward the female side and very little importance has been placed on the influence of the service bull. In this study association mapping was carried out Cor fertility trait in Holstein dairy cattle bulls using high-throughput and a high-density SNi> genotyping array. Single nucleotide polymorphisms (SNPs) were associated with dair, cattle bull fertility. Associated SNPs were queried in the bovine genome. Seven SNPs were found within the genes and fourteen were within 10 kh o! a gene. Seven gl'nes. namely LEPRELl, MOBKL3, CD247, LRRC8J\, LRFN5, IT] I [J and [·.NTP[) J were seleeted as candidate genes. Resequencing and fine mapping of selected candidate genes were performed and identified SNPs were associated with dairy cattle hull fertility. This is the first GWA study for dairy bull fertility using the Illumina Bovine S~ P50 Bcadchip containing 54001 S Ps powered by I1lumina lnfinium-Il assay. Availability: Items available for loan: UVAS Library [Call number: 1354,T] (1).

38. Preparation Of Turnip Peroxidases And Its Application To Remove The Phenolic Content Of Sannerty Effluent

by Muhammad Usman Amin | Dr. Abu Saeed Hashmi | Miss. Faiza Masood | Mr. Tanveer.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2011Dissertation note: Peroxidases are heme-containing oxidizing enzymes, which are wide spread in nature. They have the ability to catalyze the oxidation of many organic and inorganic electron donor substrates through a reaction with hydrogen peroxide or organic hydrogen peroxides. In this study peroxidase were purified from turnip using ammonium sulphate precipitation, poly ethylene glycol precipitation and zinc sulphate precipitations in order to find some simple and less expensive procedure for partial purification of peroxidases. Ammonium sulphate and PEG (6000) in the presence and absence of NaCl were used to make aqueous two phase system. Aqueous two-phase system (ATPS) without NaCl purified enzyme most efficiently. (NH4)2SO4 layer was subjected to dialysis and for further purification on sephadex gel which gave maximum enzyme activity of 1544u/mg protein. SD-PAGE analysis was done to determine enzyme purity. Purified enzyme was charged into the tannery waste water along with H2O2 to remove toxic phenolic content up to 98.24%. Availability: Items available for loan: UVAS Library [Call number: 1356,T] (1).

39. 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.

Material type: book Book; Format: print ; Literary form: drama 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).

40. To Study The Extraction, Purification And Characterization Of Papain Form Carica Papaya

by Hafiz Anis-Ur-Rehman Tariq | Miss Faiza Masood | Dr. Abu Saeed Hadhmi | Mir.

Material type: book Book; Format: print ; Nature of contents: biography; Literary form: Publisher: 2011Dissertation note: In this piece of research work, the Extraction, Purification and Characterization of Papain (a proteolytic enzyme) from Papaya fruit (Carica Papaya L. of Caricaceae family) were carried out. For this purpose, the Peelings, Flesh and Seeds of the Papaya fruit were used. Extraction of Papain was done using 0.1M alkaline Phosphate buffer. Purification of Papain was carried out by Ammonium Sulphate precipitation and dialysis followed by Gel filtration by Sephadex G-75. Then Characterization of Papain such as protein estimation, determination of proteolytic activity (International Unit) of enzyme and SDS-PAGE analysis were performed to determine the molecular weight. Finally, the yield and proteolytic activity of the Papain obtained from the Peelings, Flesh and Seeds of the Papaya fruit was measured and compared with the commercial product available in the market. Availability: Items available for loan: UVAS Library [Call number: 1362,T] (1).

41. Single Nucleotide Polymorphisms In Pouifi Gene And Its Association With Milk Production Traits In Pakistani Cattle

by Sadia Munir | Dr. Asif Nadeem | Dr. Abu Saeed | Dr. Muhammad Wasim.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2011Dissertation note: In farm animals, the primary focus of selection has been to improve milk yield. Milk field is a polygenic trait and a few potential candidate genes have been recognized. Association studies have shown that POUlFl is related to many production traits in domestic animals and is one of those candidate genes that are involved in milk production. POUl F 1 encodes a pituitary-specific transcription factor. It is well established that growth hormone (GH) released from pituitary gland plays an essential role in growth, mammary gland development and lactation process. The bovine POUlFl gene is of 15952 bp length having 6 exons. The genetic characterization of the POUlFl gene to identify the SNPs as genetic markers and validation of these potential markers by associating them with milk production traits has been performed. A total 35 samples from Sahiwal and 30 from Holstein-Friesian cattle breeds were sequenced for all 6 exonic portions of the POUlFl by using 6 sets of primers. A total 15 polymorphic sites in Sahiwal and 14 in Holstein-Friesian were identified from these sequences. Out of total 15 SNPs identified in Sahiwal, 12 were in intronic region and 3 were in exonic. Out of 14 SNPs identified in Holstein-Friesian, 10 were in intronic and 4 were in exonic region. The sequences of the amplified POUl F 1 gene fragments were aligned with the help of BLAST for SNPs identification. This study is first step in finding some confirmed markers for milk yield in Sahiwal and Holstein- Friesian cattle breed that can be used in future for selection and breeding programmes. Availability: Items available for loan: UVAS Library [Call number: 1373,T] (1).

42. Sequence Analysis Of Shiga Toxin 1 And Shiga Toxin 2 Genes Of Escherichia Coli O157: H7 Isikates From Lahore

by Saqib Hussain | Mr. Tanveer Hussain | Prof. Dr.Masroor Elahi Babar.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2011Dissertation note: Escherichia coli is normal inhabitant of all the animals and human beings. The Sorbitol non fermenting E. coli strains were detected in milk, beef and fecal samples collected from different areas of Lahore. White colored colonies of each positive sample on Sorbitol MacConkey's Agar (SMA) contained gram negative, rods with round ends non spore former Escherichia coli. Each of the isolates was sorbitol non fermenter, lactose fermenter, indole positive, Methyl Red positive, Voges Prauskaur negative and citrate negative. Each of the isolate was further characterized using polymerase chain reaction (PCR) for the presence of shiga toxin 1 and shiga toxin 2 genes. Bacterial DNA was extracted easily by boiling method when the isolates were grown on Sorbitol MacConkey's agar at 37°C for 12-24 hours. The DNA was recovered when the culture was boiled for 5-10 minutes. The isolated DNA when amplified using Stxland Stx2 specific primers showed that 68.5 percent samples were positive for Stxl and 54.2 percent for Stx2. The stx 1 and stx2 PCR products were subjected to sequencing. The resulted sequences when aligned with the reference sequence through Basic local alignment tool it showed that the shiga toxin 1 and shiga toxin 2 gene sequences are conserved and showed high similarity in their nucleotide structure. Despite of having high similarity in their nucleotide structure some haplotypes were also obtained showing single nucleotide polymorphism. Phylogenetic analysis made among local isolates and also with reported sequences from all over the world by using bioinformatics software to see the genetic similarities and difference between them. The data produced showed some highly conserved sequences and SNPs as well that will be quite useful for further applications in diagnostics and biotechnology applications in future. Availability: Items available for loan: UVAS Library [Call number: 1375,T] (1).

43. Genetic Diversity And Differentiation Of Domestic Buffalo Of Pakistan Through Sky And Zfy Genes Of Y Chromosome

by Muhammad Mudassar Manzoor | Mr. Taneer Hussain | Mr. Muhammad Asif | Prof. Dr. Abu.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2011Dissertation note: Livestock sector plays a vital role in the economy of Pakistan. Main contribution of milk comes from buffaloes and cows. Water buffalo (Bubalusbubalis) is one of the major elements of livestock in the country and possess great importance for economy in the form of milk and meat productions. Nili, Ravi,Nili-Ravi, Kundi and Azakheli are major breeds of water buffalo to be found in different areas of Pakistan. Conventional classification of breeds was based on phenotypic traits. In some cases, recent genetic studies have found differences in the structure proposed. In buffalo ,one has to bear in mind that morphological changes were not the result of adaptation to the environment, but have a social significance thus may not be indicative of the genetic relationship. In recent years Y chromosomal genes have proved to be very useful for the determination of genetic relationship among population. Comparative studies have highlighted the advantages of the SRY and ZFY genes of Y chromosome.These genes have been considered as competent and powerful tool for the purpose of breed characterization and species identification of buffaloes. In livestock sector, water buffalo (Bubalusbubalis) has shown great prospective in numerous Asian countries including Pakistan. Unfortunately, there is lack of genetic data of different buffalo breeds like Nili-ravi and Kundi which needs to be established for their genetic identification.Blood samples from true representative animals of each of the two buffalo breeds (Nili-Ravi and Kundi ) were collected from different Government livestock farms and their respective home tractsin Punjab and Sindh respectively. DNA was extracted by inorganic method and amplification of the SRY and ZFY(exon 5) genes of Y chromosome was done with especially designed primers using Primer3 software in Molecular Biology and Genomics Laboratory at Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore.Specific primers are designed for these genes amplification. Then primers were optimized for successful amplification with minimum reagent concentration. PCR was performed for amplification of SRY and ZFY(exon 5)genes on each sample. Sequencing was conducted on amplicons to find out the different single nucleotide polymorphism (SNP) to make haplotypes with the help of bioinformatics software like Blast 2sequence and Neighbour Joining phylogenetic tree was constructed by using MEGA version 5 (Tamuraet al. 2011). The results obtained from this study now can contribute to the establishment of routine DNA typing service to the advantages of the buffalo in livestock industry. Availability: Items available for loan: UVAS Library [Call number: 1378,T] (1).

44. Molecular Characterisation And Antibiotic Resistance Of Avian Salmonella Enterica

by Sajid ul Hassan Qureshi | Dr. Atif Hanif | Prof. Dr.Masroor Elahi Babar.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2012Dissertation note: Avian Salmonella enteric sub species enteritidis can cause enteritis in a wide range of host species and being responsible for the majority of Salmonella food-borne enteritis in man worldwide. This food borne disease of primary concern in developed, as well as developing countries. The spread of disease is favored by a variety of animal reservoirs and a wide commercial distribution of both animals and food products. This disease is among one of the major public health problems in terms of socio-economic impact. However there was little information on their prevalence, characterization and antibiotic susceptibility in Pakistan. In the current study Salmonella enteritidis was characterized on the basis plasmid encoded antibiotic resistance and Whole cell protein profiling to reduce the public associated with consumption of infected products. Ten Isolates were tested for plasmid encoded antibiotic resistance against six antibiotics. results of in vitro susceptibility by standard disc showed that all isolates were highly resistant to Ampicillin (100%), followed by Amoxicillin to which 60% isolates were resistant. High susceptibility was shown to levofloxacin, and Ciprofloxacin (90 and 80% respectively) by S. enteritidis isolates. Plasmid profiling of resistant isolates shown a large plasmid ( kb) that appears to be a serotype specific virulence plasmid. Whole cell protein profiling was done by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) which revealed results close relation among Salmonella entertidis isolates and the 78.1 and 35 kDa bands were observed to be major bands in all strains. Availability: Items available for loan: UVAS Library [Call number: 1380,T] (1).

45. Molecular Characterisation And Antimicrobial Resistance Of Human Salmonella Enterica

by Yaser Bhatti | Dr. Atif Hanif | Prof. Dr.Masroor Elahi Babar.

Material type: book Book; Format: print Publisher: 2012Dissertation note: Infectious diseases directly or indirectly are still the main cause of morbidity and mortality in the human population. Prevention and control of such diseases has been a major challenge since ages. During the last few decades, infections with Salmonella have been recognized as a major hazard to humans in most countries.In this study, ten confirmed and purifiedisolates ofSalmonella typhi were collected from Chugtai's Lahore Lab.and were grown inbuffered peptone water, tetrathionate broth and SS agar. Isolates were characterized by antibiotic resistance patterns, plasmid profiling and protein profiling. All isolates were resistant to ampicillin and were sensitive to ciprofloxacin and levofloxacin. 90% of isolates were sensitive to gentamicin and 50% were resistant to chloramhenicol, while 70% of isolates showed intermediate behaviour to amoxicillin. Single plasmid profile was observed among all resistant isolates and all isolates harboured a single heavy weight plasmid.Sensitive isolates were free of any plasmid. Isolates were grouped into six groups by SDS-PAGE. Different banding pattern was observed among groups. However, a protein of 10kDa was common in all isolates. Availability: Items available for loan: UVAS Library [Call number: 1382,T] (1).

46. Production And Characterization Of Hemicellulaase Activities From Aspergillus Flavus

by Hamna Qayyum | Ms.Faiza Masood | Dr. Abu saeed hashmi | Mr. Tanveer.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2012Dissertation note: The study was conducted with objective of optimized xylanase using local raw materials from indigenous isolate of Apergillus jlavus. The fungus was grown on different carbon sources including wheat bran, rice polishing for the production of xylanase enzyme. All four fungi produced xylanase activity in the medium containing wheat bran and rice polishing (1%) at pH 7.0 for 4 days. Maximum activity (14.3U/mL) was depicted by A. jlavus3 in medium with wheat bran. On the basis of better xylanase production A. jlavus3 was selected for further production and characterization of enzyme studies. The highest xylanase activity was achieved in cultivation with wheat bran (lS.8U/mL). A slightly higher quantity of xylanase was produced by the strain in wheat bran-supplemented medium (18.5 U/mL) followed by rice polishing (17.9 U/mL) when 3% carbon sources were used. The effect of supplementation of different source of nitrogen on xylanase activity by A. flavus was studied with 3 % carbon source. Of all the nitrogen sources investigated, yeast extract (organic source) was the most promising and the corresponding xylanase production was 19.9 UlmL (wheat bran) and 18.3 U/mL (rice polishing). Com step liquior was used to enhance the activity of xylanase which was approx. 10 % higher than that of control medium lacking com step liquior. The highest level of xylanase activity as well as extracellular protein using wheat bran was reported .Maximum xylanase production occurred at pH 5.5 and activities of enzyme were obtained at temperature 30°C for A. jlavus. The enzyme was purified by gel filtration, ion exchange chromatography. The enzyme activity was characterized on different temperatures and pH ranges. Availability: Items available for loan: UVAS Library [Call number: 1385,T] (1).

47. Alayisis Of Sodium Channel Subunit Beta-1 ( Scnib ) Mutations Involved In Generalized Epilepsy With Febrile Seizures

by Salma siddique | Dr.Muhammad Wasim | Dr. Abu saeed hashmi | Dr. Atif Hanif.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2011Dissertation note: Epilepsy is, one of the most common disorders of the brain, a common neurological condition defined by recurrent and unprovoked seizures. Epilepsy affects 50 million people worldwide, including one in every 200 children. Febrile Seizures (FS) are not thought of as a true epileptic disease but rather as a special syndrome characterized by its provoking factor (high grade fever) and a typical range of 6 months to 6 years. The patients with generalized epilepsy with febrile seizure plus (GEFS+) show febrile seizures initially, lasting beyond 6 years of age, and afebrile seizures occur with multiple types, mainly with generalized seizures but sometimes with focal seizures. Studies have shown that genetic factors play an important role in the pathogenesis of GEFS+ and other types of epilepsy. Mutations in a number of genes have been identified that leads to the various types of epilepsy. Sodium channel subunit beta-l (SCN1B) was the first gene identified to be associated with febrile seizures. However, very little work has been done on SCNl B gene in epilepsy patients, especially in Pakistan. The present study was conducted to understand the comprehensive role of SCN1B gene in GEFS+ patients. Blood samples of unrelated true representative of generalized epilepsy with febrile seizures plus were collected from psychiatry and preventive pediatrics departments of various hospitals of Lahore. DNA was extracted and amplified with specially designed primers and sequencing of the peR products was also done. Analysis of the sequences and SNPs/mutations was done with the help of appropriate bioinformatics softwares. In the present study, polymorphism analysis of human SCNIB gene isolated from healthy and diseased Pakistani individuals (suffering from neurological disorder, GEFs+) have been investigated for genetic association. Novel mutation IVS2-1 G> T in splice acceptor site of exon 3 have also been identified from Pakistani GEFS+ patients. This mutation was absent in the healthy (control) sample. This is first report of gene characterization and polymorphism of Human SCNI B gene from Pakistan. Likewise, in the last 15 years, several mutations in the genes have been identified which were associated with GEFs+. In addition to detecting new mutations and identifying new genes, further studies are required to elucidate the particular role of furtive mutations, genetic milieu, environment, or random events on the individual's phenotype. This study has opened a new avenue in medical sciences in Pakistan, which will help the scientists to work on genetic disease following the methodologies used in this study. The outcome of this study can further be used to confirm the hypotheses through animal modeling and proteomics. The mutation found in this study may add the information in gene databanks, which ultimately help the scientist to develop the gene therapies for genetic diseases. Availability: Items available for loan: UVAS Library [Call number: 1388,T] (1).

48. 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.

Material type: book Book; Format: print 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).

49. Molecular Characterization Of Local Isolation Of Staphylococcus Aureus On The Bsis Of 16S Rrna From Poulry And Their Transmission to Humans

by Muhammad Rizwan Ashraf | Mr. Muhammad Asif | Dr. Aby Saeed | Mr. Tanveer Hussain.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2012Dissertation note: Staphylococcus aureus is a widely distributed throughout the world and makes up the normal bacterial flora of skin and mucous membranes of man and animals. It is involved in suppurative wound infections in man and animals. Poultry industry has also been effected by S. aureus and causing great economic and health problems. The focus of the microbiology is to correctly identify S. aureus for the treatment of the animals. Molecular biology and biotechnology is proving a helping hand in the accurate identification of microorganisms through sequence analysis of 16S rRNA gene. The aim of this study was the molecular characterization of S. aureus from poultry and poultry farm workers through 16S rRNA analysis. Bacteria were collected from poultry and poultry farm human workers. All the samples were cultured and tested biochemically. In addition, peR amplification of 16S rRNA was performed in order to sequence the gene and further analyses through bioinforrnatics tools were performed. The aim of the study was the molecular characterization of S. aureus in poultry and humans through 16S rRNA sequencing, finding the phylogenetic relationships among S. aureus isolates and detection of zoonoses between poultry and human. 16S rRNA gene was amplified with peR primers and the sequence was compared with NeBI database reported S. aureus sequences. Resemblance was found between human and chicken isolates. Phylogenetic analyses were performed by using MEGA5 so ftware that also showed phylogenetic relationship among them. Availability: Items available for loan: UVAS Library [Call number: 1393,T] (1).

50. Srudy Of Gamma-Aminobutyric Acid A Receptor Delta Subnuit Gene Mutations Involved In Generalized Epilepsy With Febrile Seizures Plus (GEFS+) Patients in Punjab

by Iram Javed | Dr. Muhammad Wasim | Dr. Abu Saeed Hashmi | Dr. Asif Nadeem.

Material type: book Book; Format: print Publisher: 2012Dissertation note: World health organization (WHO) reports that neurological disorders affect one billion people worldwide, including 50 million affected by epilepsy. Epilepsy is a common neurological disorder characterized by recurrent, periodic, spontaneous and unprovoked seizures. Generalized epilepsy with febrile seizure plus (GEFS+) is an autosomal dominant disorder and a heterogeneous familial condition in which family members express febrile seizures initially, and then show multiple phenotypes of myoclonic epilepsy including partial or absence seizures and generalized tonic conic seizures. Molecular genetics techniques have identified various GEFS+ associated mutations in many genes i.e. sodium channel genes (SCN2A, SCN1A, and SCN1B) and some GABA receptor genes (GABRG2 and GABRD). GABAA receptors are the principal intermediaries of fast inhibitory neurotransmission in the eNS and have been frequently reported to playa significant role in a number of seizures. GABRD gene encodes the delta (8) subunit and is usually located in extrasynaptic GABAA receptors. The present study was aimed to investigate coding regions of GABRD gene for analyzing the mutations involved in epilepsy. Blood samples of unrelated true representative ofGEFS+ were collected from psychiatry departments of different hospitals of Lahore. DNA were extracted with the standard protocol and amplifications of the GABRD regions were done with specially designed primers. Later on, sequencing of target fragments was carried out. Sequences were analyzed through BioEdit software and then aligned with the help of custalW2 software. Out of 14 GEFS+ patients, only 3 were identified with a novel heterozygous transition mutation in intron 5. Further study, with much larger sample number, is required to revise the effects of this polymorphism and accurately identifying the associated factors. There is a need to explore the other gene mutations causing epilepsy in local population of Punjab and Pakistan that will ultimately help to develop genetic counseling strategies, gene therapies and prenatal diagnostic procedures for the population of Pakistan. Availability: Items available for loan: UVAS Library [Call number: 1394,T] (1).



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