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Item Expression of recombinant azurin domain from E.Coli(UMT Lahore, 2019) SUGHRA BANOIDAzurin is an anticancer protein, which is produced by Pseudomonas aeruginosa. It is also called cupredoxin (copper containing protein) having molecular mass 14kDa.It is a water-soluble protein.Whole Azurin domain is 128 amino acid long that helps in electron transport chain during denitrification process. Its p28 domain (molecular mass of 4kDa) is specifically involved in the anticancer activity. p28 due to its small size can be effective in treating cancer. This peptide directly targets on the cancerous cells without damaging any normal cells. This study involves the optimization of IPTG induction, time optimization in LB media, effect of temperature on bacterial growth and optimization of LB,YP and TB media. The results exhibit that the best concentration for the maximum production of transformed BL21 DH3 cells was at 1mM. Best time for the maximum production of transformed cells in LB media was 5 hours and maximum production of cells at different temperatures in TB, YP and LB media were maximum at 37°C. Optimization in three media exhibit that TB media showed maximum growth of transformed cells as compared to the other media.Item Pakistan council of scientific & industrial research (PCSIR)(UMT Lahore, 2019-09-16) RUKHSAR SHAHBAZItem Pakistan council of scientific and industrial research center (PCSIR)(UMT Lahore, 2019-10-16) Saeeda ShafiqItem Analysis of phytochemicals as inhibitors against neuraminidase (NA) from influenza virus a H1N1 (pdm09) using molecular docking.(UMT Lahore, 2019) AFIA NADEEMInfluenza virus commonly known as swine flu is well known virus which is infecting large number of population globally. It becomes threat to human lives. Phytochemicals are used to cope virus related, and other infectious diseases in people. The purpose of study is base on to known about phytochemicals which is used to treat influenza virus A Neuraminidase (pdm09). Total 105 phytochemicals were docked from which fourteen have binding affinity -6.3kcal/mol or more. Out of these fourteen phytochemicals only three showed good inhibitory effect on influenza A virus Neuraminidase. This study was based on computer based drug development software against Neuraminidase protein of Human Influenza A virus (pdm09). Molecular Docking is used to calculate binding affinities of phytochemicals.Item Study of antimicrobial activity and molecular docking of phytochemicals in OLEA EUROPAEA(UMT Lahore, 2019) FASIHA ARIFBacterial infections have appeared as a serious warning in population. By increase of synthetic antibiotics resistance, medicinal plants have used to treat such infections. In this research work, antimicrobial activity of Olea europaea parts was checked against bacterial and fungal strains. Agar disc diffusion, well diffusion and agar dilution methods was used for estimation its antimicrobial efficacy. Antioxidant effect of its all extracts was estimated by DPPH method. In silico the ADMET properties of phytochemicals of Olea europaea was checked by bioinformatics tools SwissADME and Protox. Only 39 compounds obeyed the ADMET properties and expressed the drug like similarities. These phytochemicals were docked with DNA gyrase subunit b from E.coli, FtsZ from B.subtilis and LpxC from P.aeruginosa via AutoDock Vina and binding affinities were evaluated. This study demonstrated that methanol and ethanol extract inhibited the all bacterial strains while they exposed no inhibition against fungal strain. Water extract did not show any inhibition. In agar dilution method inhibitory effect was observed by ethanol stem against P.aeruginosa and E.coli and ethanol leaves against P.aeruginosa. Antioxidant activity results exhibited that all extract exposed the positive results except methanol roots. Moreover, water leaves expressed the maximum antioxidant activity and ethanol roots showed minimum antioxidant efficacy. Computational study demonstrated that only all of the phytochemicals have inhibitory effect against selected bacterial targets by different binding affinities. The ADMET properties of Olea europaea phytochemicals and their binding to the bacterial proteins elucidated that they have effective inhibitory potential.Item Isolation of alkaliphiles and molecular characterization of their β-glucosidase(UMT Lahore, 2019) Imama AnsariExtremophiles are the organisms that require extreme environmental conditions for their growth and survival. Enzymes that are produced by extremophile organisms are known as extremozymes. Alkaliphiles are extremophiles capable of surviving in high alkaline conditions i.e. pH 9-10. Alkaliphiles can be extracted from neutral environment or high saline environment. Alkaliphiles produce enzymes that possess industrial applications. Alkaline cellulase has various applications in the field of industry, agriculture, fermentation etc. In this study the sample having pH 9 was collected from premium paper mills, Sheikhupura. This study includes the isolation of alkaliphilic bacteria, its DNA isolation, Amplification of its 16S rRNA sequence and Gene clean of amplified PCR product. 16S rRNA sequencing of bacteria showed that the bacterium belongs to genus Vibrio. The sequenced bacterium has 99.4% similarity with 16S region of Vibrio cincinnatiensis ATCC 35912. Phylogenetic analysis and Protein Structure Prediction was carried out to characterize the β-glucosidase gene. Phylogenetic tree for 16S rRNA showed that isolated Vibrio strain is more closely related to Vibrio cincinnatiensis ATCC 35912. Due to the unavailability of whole genome of Vibrio cincinnatiensis ATCC 35912 the phylogenetic tree for β-glucosidase gene and protein was constructed for Vibrio cincinnatiensis NCTC 12012 and its relationship was studied with other species of Vibrio. Phylogenetic tree for β-glucosidase gene showed that Vibrio cincinnatiensis NCTC 12012 is closely related to Vibrio tubiashii ATCC 19109 and phylogenetic tree for β-glucosidase protein showed that Vibrio cincinnatiensis NCTC 12012 is distinctly related to genus Vibrio campbellii.Item Green synthesis of copper oxide nanoparticles using plumaria alba, their characterization and applications(UMT Lahore, 2019) Maira Nadeem DarIn this study CuO NPs were synthesized using Plumaria alba which is a ornamental plant native to united states but can be grown in warm areas. The plant is known for many medicinal purposes and is easily available. The plants were obtained from university grounds and 2% of aqueous extract was made out of it using 6grams in 300 ml of distilled water. The various concentration of extracts was used interact with particular salt solution. The extract concentration was varied with constant salt concentration and vice versa until a point of reaction is obtained. The CuSO4.5H2O solution of 1mM was used in our study. Initially transparent color was observed which was changed to green upon addition of base and finally dark brown after 5 hours of continuous stirring at 200°C and 300 rpm. The NPsolution was stabilized and did not require any external stabilizing agent. . UV analysis for this solution was done which gave sharp peak in the range of 200-300 nm which showed the synthesis of CuO NPs. Later the sample was sent for characterization through SEM and FTIR and results were recorded. The results showed the size of NPs to be of the range of 144nm-266nm and hydroxyl, phenolic, ethyl, methyl and ketonic groups were present. Antioxidant, antimicrobial and catalytic activities were performed. Both catalytic and antioxidant showed positive results but that of antimicrobial showed negative result. This is probably because of large size of nanoparticles.Item Green synthesis of nickel oxide nanoparticles from plumeria rubra and their environmental applications(UMT Lahore, 2019) Mahwish JavedNanobiotechnology is an emerging field that is dominating every study from electronics to biology. Nanoparticles being the core of nanobiotechnology, is able to bring about treatments and cure of diseases which was not possible before because of their extremely small size and large surface area. In the present study, green synthesis of nickel oxide nanoparticles from Plumeria rubra was followed because of their effectiveness in many ways as no external reducing, capping and stabilizing agents are required. Green synthesis has lowest number of risks as compared to all other synthesis methods like chemical, fungal and bacterial as no chemicals or pathogens are involved in this production method. The characterization, anti-microbial, anti oxidant and catalytic activities of nickel oxide nanoparticles were performed in this study. SEM, FTIR, and UV-Vis spectrophotometer were used for physical characterization of synthesized nanoparticles. The anti-bacterial activity was checked against garm negative bacteria E,coli and Pseudomonas aeruginosa and gram positive bacterium Bacillus subtilis. The anti-fungal activity was checked against Aspergillus niger. DPPH radical was used for the analysis of anti-oxidant activity, which gave 73.7% of scavenging activity and methylene blue, Congo red and Triphenyl Tetrazolium Chloride were used for catalytic activity of nickel oxide nanoparticles. The catalytic activity of nickel oxide nanoparticles was positive against all dyes studied in this research.Item Study of anti-microbial activity and molecular docking of phytochemicals in asclepias curassavica(UMT Lahore, 2019) Fariha AmirPlants are widely used for many purposes. Traditionally plants are used as medicine from ancient time even when science of medicine never existed. At that time people used plants as a medicine and that was a natural way to cure any disease which was a slow process but had no side effect. Now a day’s synthetic drugs are use as antimicrobial agent, antioxidant agent and for many other treatments, which have many side effects but the major, problem is that our body started showing resistance against these drugs, due to which drugs generation increases day by day which is an alarming situation. The purpose of this study is to confirm that either the selected traditional plant shows high activity as an antimicrobial and antioxidant agent or not, if yes then what is the mechanism behind it and also to examine the toxic phytochemicals present in it which can harm human beings. The antimicrobial activity of 3 bacterial strains E. coli, B. subtilis, P. aeruginosa and 1 fungal strain A.niger were examined against the methanolic, ethanolic and distilled water extract of roots, stems and leaves of Asclepias currasavica. These extracts inhibit all bacterial strains as leaves ethanol, leaves distilled water and leaves methanol showed inhibition zone of about 12mm, 9mm and 6mm against P. aeruginosa while root methanol showed inhibition zone of 6mm against E. coli. These extracts also showed inhibition against B. subtilis, but do not showed any inhibition against selected fungal strain. The antioxidant activity of all extracts was examined by using DPPH assay. The result demonstrated that stem distilled water showed high antioxidant activity. The mechanism of action of nontoxic phytochemicals were also examined by molecular docking after checking their ADME and ProTOX properties. Out of 100 phytochemicals, only two phytochemicals were nontoxic for human health and can be used as a drug.Item Green synthesis of silver nanoparticles from plumeria obtusa and evaluation of their activities(UMT Lahore, 2019) Mohammad Akber KhanThe present research work deals with the synthetization of silver nanoparticles AgNPs by using the aqueous extract of Plumeria obtusa and monitoring was carried out at regular time intervals utilizing UV–vis spectroscopy. From the experimental analysis, the optimum conditions for AgNPs biosynthesis were found to be 1Mm AgNO3 concentration, 30 minutes sunlight exposure. SEM analysis confirmed the existence of Ag-NPs with average size ranges 81.4nm to 95.5nm. From SEM the silver nanoparticles shape was confirmed which was nanocluster. In addition to this, FTIR analysis and UV Vis was done for characterization. Thus sufficient antioxidant activity was observed in the acquired AgNPs. Ag-NPs give good catalytic activity against Congo red dye, methylene blue and TTC. Silver nanoparticles give antibacterial activity against E.coli and Pseudomonas aeruginosa and Bacillus subtilis. Antifungal activity was also checked.Item Blood spatter analysis on different types of fabrics of Pakistan(UMT Lahore, 2019) Khansa Ahmad KhanThe study of textiles damage is an immensely valuable resource in the criminal examination that is used to find a way to resolve the stabbing cases, shooting cases and rape allegations. It also can overcome arson threats and "acid assaults." Here, the literature is reviewed to examine work on blunt force impact, stabbing and slashing wounds, the occurrence of proof forgery in sexual attacks, projectile harm, and the effect of decomposition on textile damage, amongst many others. It is evident there is a shortage of work in the field, but when the study was done, it was insightful and meaningful. Nevertheless, there are fields where more research is required, such as a thorough analysis of usual wear and tear, and ability of additives to inflict harm to textiles. This research deals with blood pattern analysis on 6 different fabrics that are most common in Pakistan. Almost 8 patterns were applied on these 6 fabrics to analyze bloodstain radius, diameter, width, height, length, angle of impact and area. These all parameters were studied on both laundry and brand new fabric to analyze differences in results. It was observed that bloodstains cover most of area on laundry fabric as compared to brand new fabric. Blood drop sizes were also observed that was different according to height. Absorbance factor of fabrics were also checked and it was observed that net had high absorbance factor.Item Green synthesis of Silver Nanoparticles and their characterization(UMT Lahore, 2019) SHANZA LIAQATBackground in recent study nanotechnology is the emerging field of science in which researchers take great interest in nanoparticles synthesis. Word Nano means “small”. Nanoparticles are very small in size that range from 10-100nm in one dimension. These particles are very reactive and can penetrate in cell easily due to their small size and optical properties. There are various physical, chemical and biological methods for their synthesis. Biological method is mostly use for their synthesis because this method is environment friendly and less toxic and easy. Nanoparticles have great use in the field of physics, chemistry, bio-medical science and agriculture.Item Plant-mediated synthesis of silver nanoparticles by using chichorium intybus and their characterization(UMT Lahore, 2019) UMERA NAWAZThe present work reports plant-mediated synthesis of Ag nanoparticles i.e., formation of AgNPs using plant extract of Chichorium intybus as an ecological green synthetic approach. The reduction of silver was carried out by plant extract which not only act as a reducing agent but also serve as capping agent. The change in color was a primary indication of formation of AgNPs nanoparticles. UV-Vis spectroscopic investigation reveals the final conformation for the formation of AgNPs. The synthesized NPs have been characterized using Fourier transform infrared (FT-IR) and UV-Vis spectroscopy. Further, Scanning Electron Microscope (SEM), studies have been conducted to evaluate size, morphology and shape of NPS. The SEM images illustrated that the nanoparticles were spherical with different in size. As a result, this is very simple, cost effective and green approach for development of NPs and it is valuable in biotechnological, environmental, and biomedical applications.Item The comparison of classical and newly designed anthropometric indices in the assessment of Dyslipidemia(UMT Lahore, 2019) M. MUNEEB SOHAILIt has been shown that dyslipidemia involves low concentrations of HDL but high concentrations of other plasma lipids. This study reanalyzes the relation between plasma lipids and various anthropometric parameters. In this cross-sectional type of study, the participants (n=53) were recruited for the study. Participants were given one day free health camps and they were anthropometrically measured. Plasma lipid fractions were accordingly determined. Statistically significant and non-significant results were obtained for different lipoprotein fractions. Although BMI was a good classical indicator of obesity, but other indicators like ABSI and BRI, also proved significant in assessing the type and severity of dyslipidemia. Different lipoprotein fractions behaved differently with anthropometric parameters. Our study includes 79% of dyslipidemic while 21% of non-dyslipidemic patients. Males were more inclined towards lipid abnormalities than females especially HDL abnormality. Abnormal HDL level was mostly associated with rural population. But consumption of fast food may prove fatal for lowering the HDL level in urban population also.Item Study of antimicrobial activity of the extracts obtained from the alcea setosa leaves and flowers(UMT Lahore, 2019) Anwal RazaThe increased resistance of microorganisms to the currently used antimicrobials has led to the evaluation of other agents that might have antimicrobial activity. Medicinal plants are sources of phytochemicals which are able to initiate different biological activities including antimicrobials. The use of medicinal herbs in traditional system of medicine is a common practice in many cultures around the world especially in Asian societies. Alcea setosa plant is belong to a family Malvacae and is found in different regions of the world. The Alcea setosa plant is used to treat cough, chest pain, intestinal inflammation, respiratory diseases, pectoral diseases, wound healing and female urinary system disorders. It flowers and leaves are also used as a skin laxative, demulcent, diuretic and emollient. The present study was carried out to evaluate the antimicrobial property of Alcea setosa leaves and flowers against certain bacterial strains. The test organism were Bacillus subsitlis, Pseudomonas aeruginosa and Escherichia coli. The antimicrobial activity of Alcea setosa was checked by two different methods, the disc diffusion method and dilution method. All the different extracts (methanolic, ethanolic and aqueous) of leaf and flowers exhibited the significant anti-microbial activity against different strains. Results showed that leaf ethanolic extract showed highest zones of inhibition against Bacillus subtilis, while leaf methanolic shown highest inhibition against E.coli. Similarly, flower ethanolic and flower methanolic and leaf ethanolic showed zones of inhibition only against Pseudomonas aeruginosa in dilution method. The phytochemical test shows that the flavonoids, steroids, terpenoids, tennis, saponins, phlobatanin and alkaloids are present in the plant extracts. These results therefore justify the traditional use of Alcea setosa leaves and flowers to treat different diseases.Item GC-MS analysis of methanolic extracts of Alcea Setosa (leaves and flowers)(UMT Lahore, 2019) Aqsa FayyazPlants are the traditional sources for many chemicals used as pharmaceutical biochemical in many countries. Alcea setosa is an ornamental plant which belongs to flowering plant family Malvaceae and commonly known as bristly hollyhock. The flowers and leaves of Alcea setosa are used in treatment of many diseases such as inflammation, stomach ailment and breathing illness. The major aim of study was to investigate the bioactive compounds of Alcea setosa flowers and leaves by GC-MS analysis. The chemical composition of the Methanolic extract of Alcea setosa parts was done by standard protocol using the equipment Clarus 500 Perkin – elmer (Auto system XL) Gas Chromatograph- Mass Spectroscopy. GC-MS analysis of the flower extract revealed the existence of seven compounds after matching the mass spectra with NIST library. While eleven compounds were identified from leaves extract. The major compounds present in flower Methanolic extract were 2-O-(2,2dimethylpropyl) 1-O-(4-methylpentyl) oxalate, 2-ethylhexyl nonyl sulfite, 2,3epoxyhexanol and N-(5-chloro-2-hydroxyphenyl) dodecanamide. The vital compounds present in leave extract were Spathulenol, Octacosane, Hexanedioic Acid, Dioctyl Ester and Heneicosanoic Acid, Methyl Ester. The components of extracts of Alcea setosa could be responsible for many medicinal and antimicrobial activities.Item Lahore general hospital department of Pathology(UMT Lahore, 2019-10-15) Rubab BatoolItem Isolation and screening of mannanase producing bacterial co-cultures from soil using submerged fermentation(UMT Lahore, 2019) Sadia TabassumEndo-1,4-β-D-mannan mannohydrolase cleaves the backbone of β-1, 4 mannosidic linkages in mannan chains and releases linear/branched oligosaccharides of various length. In plants, mannan is the dominant hemicellulosic polysaccharide having β-1, 4-linked backbone that contains mannose or a mixture of mannose and glucose residues. Mannanase play a vital role in the paper, pulp, food, feed industry, and considered as potential for biofuels production. Besides other sources, microbial mannanase are preferred because the presence of all desired characteristics for industrial applications. This study aims to isolate and screen high potential mannanase producing bacterial strains from soil samples and produce mannanase from bacterial co-culture using submerged fermentation. Minimal agar media was used to isolate mannanase producing bacterial strains. The strains were sub-cultured and preserved on Nutrient agar media. After that, the preserved strains were tested for their compatibility and then were further screened using submerged fermentation. After fermentation, enzyme assay was performed. The strains of bacterial co-culture no.11 shown maximum absorbance at 560 nm were further identified by morphological and biochemical tests. It was shown that the strain 1 of bacterial co-culture no.11 was identified as bacillus rod shape, irregular and was positive for gram staining, Voges Proskauer test and catalase test and negative for endospore staining. As for strain 2, it was observed as cocci and it was positive for gram staining, endospore staining, catalase test and Voges Proskauer test. From this study, it was concluded that strains of bacterial co-culture no.11 can important for the production of mannanase as this co-culture released maximum mannose sugar in the solution. The results need to be confirmed at large scale for statistical analysis.Item Production of 1, 4-β-D Glucanase from bacterial co-culture using solid-state fermentation(UMT Lahore, 2019) Nazish Sajjadellulase or 1,4-β-D-glucan 4-glucanohydrolase is an enzyme that cause the hydrolysis of cellulose or 1, 4-β linkages into simple glucose monomers. It is used worldwide because of its extensive role in food, wine, textile, laundry, pulp and paper industry, animal feed, agriculture, biomass, and refining industries. Cellulases form plants and animals are more expensive and cannot be readily used in industries so microbial cellulases are preferred over them because they can grow on environment friendly and cheap substrates. The present study deals with the isolation of cellulolytic bacterial strains and cellulase production from bacterial co-culture. Seven different bacterial strains were isolated from the soil, screened on cellulose agar media and further tested for their compatibility. Secondary screening was done for the bio-synthesis of enzyme using Solid state fermentation (SSF). Out of 10 bacterial co-cultures, co-culture no. 3 shown maximum yield of cellulase (514 U/ml) using 10g of solid substrate. The strains of co-culture no. 3 were further identified using morphological and biochemical tests. It was observed that both strains of bacterial co-culture no. 3 were gram-negative, strain 1 was rounded and strain 2 was rod-shaped. Both strains shown positive results for catalase, methyl red and endospore staining tests, but for voges proskauer strain 1 gave positive test. From this study, it was revealed that bacterial co-culture no.3 can be the high potential co-culture for maximum cellulase production on large scale.Item Computer aided investigation and molecular docking of phytochemicals as inhibitors against brain-eating amoeba (Naegleria fowleri)(UMT Lahore, 2019) SamanNaegleria fowleri is pathogenic for humans. This infection grows very fast and causes the neural damage of the patient. Phytochemicals are acknowledged to have high potential to eradicate bacterial, parasitical and viral infections in human beings. Phytochemicals have been used for the cure of infections in human for long-time and these phytochemicals produce naturally in Pakistan. It was found that from them only Prenylicoflavone A, Phinflavanone and 3a-dihydrocadambine showed effective result for Calcineurin B (Nfa 2 gene). IsoangustoneA, PrenylicoflavoneA and Shinflavanone showed effective results for Hemerythrin like protein (Nfa 1 gene). Prenylicoflavone A can be the best candidate to use as a drug for N. fowleri infection because this phytochemical showed significant binding affinity both for Nfa 1 and Nfa 2 gene. Nfa 2 showed greater binding affinity value than Nfa 1 confirming that Nfa 2 is a good target for drugs than Nfa 1. This study was designed at computer based drug development software against Hemerythrin like protein (Nfa 1 gene) and Calcineurin B (Nfa 2 gene) of N. fowleri. Molecular docking based analysis was performed to describe the binding affinities of different phytochemicals.
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