1.
Molecular Approach For Sex Determination In Avian Species
by Sehrish Basheer | Dr. Asif Nadeem | Dr. Muhammad Wasim | Madam.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2011Dissertation note: To tell the difference between male and female birds takes keen observation. While not all bird species have easily visible gender differences. Almost all birds' species are sexually monomorphic so it is difficult to distinguish between male and female characters. There are some techniques for bird's sex identification. Surgical Sexing, in this an endoscope is passed into the body cavity and ovaries or testis can be observed. It is available for all types of birds. Surgical sexing is oldest and quickest method. But it cause stress to the bird and expensive technique. Fecal steroid analysis is another technique in which stool sample is analyzed for reproductive hormone presence. In this technique bird should be sexually mature for analysis. Bload Sexing is used to determine the male or female bird. This can be done at any age. Feather exing is very useful and economical method. Retrieve a feather pulp from growing feather and analyze the presence of male and female chromosome. Two chromosomes Z and Ware present in birds. ZZ in male and ZW in female. The method used in this study was based on the chromosome differences. Nine different species of birds which includes green parrot, budgerigar, .pigeon, quail, sparrow, chicken, peacock, duck and pheasants were analyzed and their sex is determined by molecular methods. DNA will be extracted from feathers and blood. The intronic egion of CHD 1- Wand CHD l-Z gene will be amplified by sex specific primer. PCR products were screened by agarose gel electrophoresis. The PCR products were show double and single bands on the agarose gel. The double bands indicate female bird because of ZW chromosome is present in females and single band indicates the male bird because of ZZ chromosomes in males.
Availability: Items available for loan: UVAS Library [Call number: 1400,T] (1).
2.
Comparison Of Locally Available Synthetic And Non-Synthetic Powders For Latent Fingerprint Development
by Arman Khan | Dr. Muhammad Wasim | Dr. Abu Saeed Hashmi | Dr. Wasi.
Material type: ; Format:
print
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1718,T] (1).
3.
Estimation Of Caffeine In Decaffeinated Coffee And Tea Available In Pakistan
by Muhammad Abbas Sadiq | Dr. Muhammad Wasim | Dr. Abu Saeed Hashmi | Ms. Sehrish.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1719,T] (1).
4.
Estimation Of Various Adulterants In Milk Available In Local Market
by Farhan Tanveer | Dr. Muhammad Wasim | Dr. Abu Saeed | Ms. Sehrish Firyal.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1781,T] (1).
5.
Bioconversion Of Molasses To Glucose Oxidase Through Solid State Fermentation With Aspergillus Niger
by Wajeeha Zafar (2012-VA-574) | Dr.Abu Saeed Hashmi | Dr. Muhammad Tayyab | Dr. Muhammad Wasim.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Enzymes can be defined as soluble colloidal organic catalysts which are produced by living cells and are capableof acting independently of the cells. Glucose oxidase belongs to oxidoreductaseand is also called as glucose dehydrogenase. The glucose oxidase enzyme (GOX) oxidizes glucose to gluconic acid. In cells, it aids in breaking the sugar down into its metabolites. Glucose oxidasehas found several commercial applications including glucose removal from dried egg; improvement of color, flavor, and shelf life of food materials; oxygen removal from fruit juices, canned beverages. It has also been used in an automatic glucose assay kit in conjunction with catalase and chiefly in biosensorsfor the detection and estimation of glucose in industrial solutions and in body fluids such as blood and urine. It is often extracted from Aspergillusniger. GOX is a dimeric protein. The active site where glucose binds is in a deep pocket. This enzyme acts outside of cells, is covered with carbohydrate chains (Raba and Horacio 1995).
Aspergillus niger is the potential source for the production of glucose oxidase and is preferreddue to its high production ration of extracellular enzyme. The ability of Aspergillus niger toutilize a wide range of waste products as nutrition source makes it more economical source of the enzyme (Rajesh et al .2002).
The glucose oxidase fromA. nigerisalso an intracellularenzyme present in the mycelium of the organism. Aspergillus nigeris a filamentous fungus belonging to phylum Ascomycota. It Produces microscopic conidia on conidiophores that are produced asexually. Hyphae possess septa and are hyaline. They are supported at their base by foot cells from which conidiophores originate. It possesses long, double-walled, smooth and colorless to brown conidiophores.It is commonly foundin mesophilic environments such as soil, plants and enclosed air environments. It is capable of surviving in various environments, it is not only a xerophilic fungus, but is also a thermo tolerant organism. It is because of this property that it exhibits a high tolerance to freezing temperature(Schuster 2002).
Glucose oxidase was first isolated from mycelia ofA. nigerandPenicilliumglaucumby Müller.
A large number of microbes including bacteria and filamentous fungi have been used for the production of glucose oxidase. Glucose oxidase is produced at large scale using A. nigerand P. amagasakiense. Many bacteria are also involved in the production of this enzyme; some of these are Zymomonasmobilis, Micrococcus and Enterobacte(Yogananth et al. 2012).
Glucose oxidase (GOX) from Aspergillus niger is a well-characterised glycoprotein consisting of two identical 80-kDa subunits with two FAD co-enzymes bound. Both the DNA sequence and protein structure at 1.9 A have been determined that these identical subunits size vary from 70 to 80 KDa. It catalyzes the oxidation of D-glucose (C6H12O6) to D -gluconolactone (C6H10O6) and hydrogen peroxide. It is produced naturally in some fungi and insects where its catalytic product, hydrogen peroxide, acts as an anti-bacterial and anti-fungal agent (Ikram et al . 2014).
Glucose oxidase has a molecular weight of 160,000 a.m.u. (Tsugeet al .1975) and consists of two identical polypeptide chain subunits having nearly equal molecular weights linked by disulphide bonds (O'Malley and Weaver 1972) and it is highly specific for β-D-glucose (Bentley 1963). Each subunit of the glucose oxidase contains one mole of Fe and one mole of FAD (Flavin adenine dinucleotides) and it contains 74% protein, 16% natural sugar and 2% amino sugars (Tsugeet al. 1975). The Glucose oxidase enzyme in its purest form is pale-yellow powder. The molecular weight of GOX ranges from approximately 130 kDa to 175 KDa (Kalisz et al. 1997).
Gluconic acid, the oxidation product of glucose, is a mild neithercaustic nor corrosive, non-toxic and readily biodegradable organic acid of great interest for many applications. As a multifunctional carbonic acid belonging to the bulk chemicals and due to its physiological and chemical characteristics, gluconic acid itself, its salts (e.g. alkali metal salts, in especially sodium gluconate) and the gluconolactone form have found extensively versatile uses in the chemical, pharmaceutical, food, construction and other industries (Anastassiadis and Morgunov 2007 ).
This enzyme is present in all aerobic organisms and normally functions in conjunction with catalase (Coxon and Schaffer 1971). This enzyme is also used as an antioxidant (Berg et al. 1992). It is mainly available from microbial sources and is normally produced by aerobic fermentation of Aspergillus nigerand Penicillium species (Fiedurak1996; Lu et al. 1996; Plush et al .1996; Rando et al. 1997). It has high specificity for D-glucose (Kuly and Cenas 1983).
This enzymeis also widely used to produce gluconicacidthatGOXtogether with Horse Reddish peroxidase has a range of applications in the food industry for glucose determination. GOX is being used in the textile industry producing hydrogen peroxide for bleaching process. This enzyme is also used to determine capillary glucose in screening of gestational diabetes (Mesiggi et al. 1988). This enzyme is utilized to extend the shelf life of fish(Field et al.1986) andproduction of calcium gluconate, gluconic acid and its derivatives (Khurshid2009).
Solid state fermentation [SSF] has been recently considered as the most cheapest and more environmentally friendly relative to submergedliquid fermentation [SLF] in the production of value added industrial based products such as enzymes, bio fuels.Advantages of Solid State Fermentation over Submerged Fermentation isHigher volumetric productivity, usually simpler with lower energy requirements, Might be easier to meet aeration requirements, Resembles the natural habitat of some fungi and bacteria and Easier downstream processing (Mienda et al. 2011).
Molasses is a dark brown, almost black, moist granular sugar. Its distinctive molasses taste is due to its high content of minerals. Nutritively, it has high iron content (Draycott and Philip 2008).
Aspergillus niger is a fungus, one of the most common species of the genus Aspergillus.The genus Aspergillus isimportant economically, ecologically and medically (Nizamuddinet al.2008).
Glucose oxidase enzymewas producedthrough the microbial fermentation. For that purpose solid state fermentation wasdevelopedwithA.niger. Solid state fermentation was applied to utilize agricultural residue such as Molasses as substrate.
The current research work was focused on production ofextracellular Glucose Oxidase (GOX) fromAspergillusniger using industrial waste such as molasses as substrate by Solid state ( static ) fermentation.
Availability: Items available for loan: UVAS Library [Call number: 2219-T] (1).
6.
Detection And Analysis Of Improvised Explosive Devices Used In Terrorism Activities In Pakistan
by Arslan Nazar (2012-VA-630) | Dr. Sehrish Firyal | Dr. Muhammad Sarwar | Dr. Muhammad Wasim | FaizaMasood.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Defence and security organizations are in steadyrequirement of finding new options for the detection of explosives. Fundamental applied research in this area focuses on uncovering of highly energetic substances as well as home-basedexplosives that could be a weapon of mass devastation (Marshal and oxley, 2009 yinon and Zitrin 1996, Scubert and Rimiski-Korsakove, 2006). Current methods of detection for explosives or highly energetic materials are based on a wide variety of technologies that focus on either massexplosives or little portions of explosives. Mass explosives can be distinguishedin some way by imaging features,character of the explosives charge, wires and detonators or unswervingly by spotting the chemistry and dielectric characteristics of the explosive substance. Trace recognition method relay on revealing of vapours given off by the explosives or on explosive’sconstituent part that are set down on nearbyexteriors (national Academy of sciences Committee, 2004). Even though, numerous published material is available about methods of sensing of explosives present in air,water, clothing,soiletc and these put forward the benefit of providing traceconfines of sensation at ppb intensities (Caron et al,. 2010; Hilimi and Luonge). Inthe bulk of the criminal acts, sampling is done at the scene trailed by a sample preparationmove, to be shortly processed by a particular technique for analysis. Sampling and samples preparation are amidmajor, shortcomings in explosive uncovering in many cases frightening the physical condition and life of examiner and the responding officer.
Improvised explosive devices are widely used by military in wars and police to keep up regulation and command. Gush of terrorist activities and increase in criminal conduct have been a matter of great concern worldwide and particularly for Pakistan. There have been motiveless
annihilation of private and community properties as well as industrial centres, causing irretrievabledamages to state and local markets and imperillinghumanexistence(Shen et al. 2005). Potentially perilous explosives like dynamite, varied military explosives havingnitroglycerine (NG), Cyclotrimethylenetrinitramine commonly known as RDX, Cyclotetramethylenetetranitramine and alternativehome-produced low explosives and provocative devices are now currently promptly obtainable to scandalous and terrorists. The haphazard and deliberate uses of these explosives consist ofextortion of cash and taking vengeance, unlawful transportation of prohibited substances, assassinations, terrorist and delinquent activities in numerous regions of the country (Sharma and Lahiri 2005).
Recognition of detonating method, estimating the path ways taken by explosive transportation andarresting the anti-social charactersconnected with unstable materials and explosions is primary aim of explosive analysis. For this purpose various explosive substances and explosive remains are to be examined qualitatively and the ingredients are to be approximated quantitatively using primarily by thin layer chromatography (TLC). TLC is a technique employed for the screening of organic constituents at hand in the post blast samples. The identification of explosives containing alkylammonium nitrate is done by TLC. Secondly Gas chromatography with mass spectrometry (GC-MS) technique with the benefit of anelevated resolving supremacy is avitalapparatus for the analysis of chemical composition of explosives. The extremelyproficient GC analysis withcapillary columns authorizes the examination of explosive hydrocarbons, identical substances of nitroaromatics, hexogen (RDX) and the high explosive pentaerythritoltetranitrate (PETN) in a single run. The spectroscopic identification of explosive materials by FTIR is striking due to the intrinsicpotential of real-time detection, non-vicious analysis, and nominal sample preparation, thirdly the scanning electron microscope (SEM) produces an increased image of the sample based on the contact of an electron beam with the sample’s exterior. Finding of minutemasses of explosive remains play an important part in forces, inhabitant, and counter terrorism requests(Pacheco-Londono et al. 2005). To press on explosives sensor methods, present methods need to become affordable and transportable without disturbing the integrity of the devices. The uncovering of ordinary explosives as well as trinitrotoluene (TNT), RDX, HMX, 2,4,6 Trinitrophenyl-N-methylnitramine (TETRYL)Pentaerythritoltetranitrate (PENT), and NG were carried out using diverseprocedures(Sanchez et al. 2007).
Detection of explosives is anparticularly relevant analytical concern for law enforcement personals and for the environmental protection agencies. As the use of explosive substances have been increased by the terrorists, problems have increased for law enforcement and environment and security agencies regarding the detection ofexplosives residues in baggage, parcels vehicles, aeroplane, on travellers, etc. In bomb scene investigations, it is important to find debris that includes detection of explosive residues. Mobile and hand held explosives detectors, similar to those used for detecting hidden explosives, can be of great help in detecting such residues. Several methods i.e. GC/MS, SEM, FTIR were used in Punjab Forensic Science Agency (PFSA) to analyze residues of explosives. The detection of landmines is an acute, urgent worldwide problem that needs specific and effective detection methods (Yinon 2002). Keeping in mind the above said situations, the project was designed with following objectives
Availability: Items available for loan: UVAS Library [Call number: 2235-T] (1).
7.
Mutational Screening Of The RB1 Gene In Pakistani Patients With Retinoblastoma
by Saeeda Kalsoom (2007-VA-555) | Dr. Muhammad Wasim) | Dr. Khushnooda Ramzan | Dr. Ali Raza Awan | Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Retinoblastoma is a neonatal intraocular tumor caused by biallelic inactivation of RB1 gene. Rb
patients and asymptomatic carriers undergo a series of clinical tests for diagnosis and tumor
treatment. These clinical examinations prove to be expensive and time consuming. On the other
hand if the proband’s RB1 gene mutation status is determined by genetic testing, it can prove as
more significant and cost effective diagnostic methods. Secondly, only those asymptomatic or at
risk carriers with the mutation, require clinical surveillance while those proven to be unaffected
do not require additional clinical examinations. Furthermore early diagnosis of Rb by molecular
testing can enable and enhance clinical management, earlier treatment, follow-up care, carrier
screening, genetic counseling, prenatal diagnosis and reproductive planning in predisposed
families. Irrespective of the importance of molecular testing of Rb patients, in Pakistan only a
few clinical reports on Rb are available so, there was a dire need to find RB1 mutations in
Pakistani Rb patients and to set a molecular based diagnosis for poor affected families. Keeping
in view the importance of molecular diagnosis, in this study a reliable genetic test has been
developed to detect the RB1 germline mutations in Pakistani Rb patients.
During this study, 70 Rb patients including 38 unilateral and 32 bilateral cases were enrolled,
from different regions of Pakistan. By using direct sequencing method, seven novel and twelve
reported RBI mutations were found. The novel mutations included three frameshift mutations
(c.1116_1119delCACT in exon 11, c.1436_1437delAC in exon 16 and c.2060_2061insTCATT
in exon 20) and four substitutions (c.148G>T in exon 2, c.610G>T in exon 2, g.94G>C in exon
7, c.947A>T in exon 10 and g.1991G>C in promoter region) while twelve reported mutations in
146
22 patients included, 9 substitutions (c.160G>T in exon 2, c.289G>T in exon 3, c.751C>T in
exon 8, c.920C>T in exon 9, c.967G>T in exon 10, c.1072C>T in exon 11, c.1654C>T in exon
17, c.2063T>C in exon 20 and c.2359C>T in exon 23), one frameshift mutation (c.772_776del in
exon 8) and two splice site mutations (c.380+1G>T and c.1215+1G>A in intron 3 and 12
respectively). Mutation detection rate was found to be 77.8% in (7/9) bilateral familial, 50% in
(2/4) unilateral familial, 56.5% in (13/23) bilateral sporadic and 14.7% in (5/34) unilateral
sporadic patients while overall rate of mutations in bilateral and unilateral patients was detected
as 62.5% (20/32) and 18.4% (7/38) respectively. Beside mutations one novel c.940-64C>T
(intron 9) and nine reported intronic variants c.380+45 C>T (intron 3), c.501-77G>A (intron 4),
c.1128-72T>G (intron 11), c.1695+99A>T (intron 17), c.1695-1696delAA (intron 17), c.1815-
104A>G (intron 18), c.1961-10T>C (intron 19), c.2663+33T>C (intron 25) and c.2664-10T>A
(intron 25) were also found. Carrier screening facility was also provided to six asymptomatic
siblings (as possible carriers) of familial proband but none of them was found to be diseased.
Hopefully, in future the findings and developed protocol of this study will help to reveal the
molecular basis of Rb in Pakistani Rb patients which additionally help to secure vision and life
of Rb patients. Further, in Pakistan there is dire need to develop “National Rb Registry Centre”,
to register all new Rb cases for finding incidence rate and prevalence of Rb in Pakistan. Beside
this other related issues like financial constraints, health education, planning and awareness
about Rb, occupational training for health providers, capacity building for neonatal
ophthalmologic screening and cosmetic rehabilitation for surviving Rb patients are important and
should consider. Availability: Items available for loan: UVAS Library [Call number: 2370-T] (1).
8.
Genetic Identification And Characterization Of Pakistani Birds Of Perdicinae Subfamily (Partridge) Through Dna Barcoding Method
by Asim Iqbal Jutt (2013-VA-557) | Dr. Ali Raza Awan | Dr. Muhammad Wasim | Prof. Dr. Kamran Ashraf.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Pakistani birds of Perdicinae sub family are cage and game birds. Birds includes Altectoris chukar, Ammoperdix heyi, Ammoperdix griseogularis, Francolinus francolinus and Francolins pondicerianus. Traditional methods of identification were based on the phenotypical characterization of birds, which may lead to incorrect identification, so there was need to explore their characters at DNA level for accurate identification and to establish a DNA reference.
Birds of sub-family Perdicinae have not been genetically characterized in Pakistan. A new precise method “DNA barcoding” was applied using COI gene of mDNA for authentic identification and classification of these birds. Blood and tissue samples of five species (fifteen samples) were obtained. DNA of each sample was extracted by organic method. Amplification of CO1 gene was done by using a universal set of primers BIRDF1, BIRDR1. Sequence of 450bp were analyzed using bioinformatics softwares. Each sample was aligned with its reference sequence of COI gene available on NCBI. Every nucleotide position which did not align with the reference sequence was studied to identify SNPs. A common phylogenetic tree of all partridges showed that they have common ancestor about 0.7 million year ago, F.francolinus, F.pondicerianus and A.heyi shared a common clade whereas A.chukar made a separate clade from the ancestor. A.heyi and F.pondicerianus showed closed resemblance. It has been proved that DNA barcoding is an efficient and accurate molecular tool for species identification and phylogenetic implication. This study established a DNA Data Bank that helped scientists to investigate the biodiversity, taxonomic classification, species identification and also established foundations for molecular biologists to study taxonomic uncertainties at sub species level using SNP based identifying marker. Availability: Items available for loan: UVAS Library [Call number: 2714-T] (1).
9.
Polymorphism Analysis Of Bovine Tumor Necrosis Factor Alpha Gene And Its Association With Mastitis In Nili Ravi Buffaloes
by Samia Tanveer (2011-VA-362) | Dr. Sehrish Firyal | Dr. Muhammad Wasim | Dr. Muhammad Nawaz.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Various number of factors cause hindrance in the milk production potential of buffalos. Mastitis is the costly and most prevalent disease causing production losses of dairy herds in Pakistan and elsewhere in the world. Susceptibility and resistance to mastitis is complex trait influenced by genetic variation of animals. Among these immunity gene variations, the polymorphism in tumor necrosis factor alpha gene (TNF-α) play important role in immune response to virus. Polymorphism in TNF-α gene is associated with mastitis susceptibility and resistance. It would be a potential candidate gene for imparting resistance mastitis in dairy buffalos. Blood sample were taken from the 20 Nili Ravi buffalos having clinical and subclinical mastitis. Extraction of DNA was done from frozen blood after thawing, using organic extraction method & also kit method followed by DNA quantification (i.e. gel electrophoresis and nanodrop). Total 5 primers were designed using Primer3 bioinformatics tool. All these primers were optimized using different protocols and a set recipe was obtained for each primer. The amplification of DNA samples was done one by one using all these five primers on optimized protocol. The amplicons obtained were subjected to agarose gel electrophoresis to check whether we have the required product or not using 100 kb ladder and then amplicones were send for the sequencing.
Summary
110
The sequencing analysis of resulted amplicon sequence was done using Bioinformatics software Finch TV. Total of 6 mutations were found while 5 were same in all the samples whereas 6th mutation was found only in clinical samples. It is valuable in accomplishing genetic progress for resistance and to improve the immune response. This study will paved the way for animal breeder for selection of Nili Ravi mastitis resistant buffalos for breeding. TNF-α gene polymorphism based marker is now available for screening of resistant bulls as well. Availability: Items available for loan: UVAS Library [Call number: 2936-T] (1).
