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    ABSTRACT: Bacillus sp. RM16 was isolated from a hot spring in Karachi and screened for the production of α-amylase. The enzyme was obtained after 72 h cultivation of strain in Luria broth containing 1% starch (w/v). Enzyme Amy RM16 was purified to electrophoretic homogeneity by a series of sequential steps including precipitation with ammonium sulfate at 70% saturation, Q-Sepharose, Phenyl Sepharose and reversed phase chromatography. The purified enzyme is made up of a single polypeptide chain of 66 kDa as established by a combination of SDS-PAGE and zymographic analysis. In our experimental conditions, a total yield of 1.35% with specific activity of 6380U/mg was obtained providing 17 fold final purification of the enzyme. Biochemical characterization of the Amy RM16 such as optimum temperature and pH, substrate specificity and enzymatic susceptibilities towards different metal ions and inhibitors were also performed. Results of these studies revealed that, the enzyme is active at wide temperature range with optimum activity at 80°C and retained 85% of the activity for 3 h at 50°C and around 50% of remaining activity for 1 h at 80°C. The enzyme showed optimum activity at pH 5.0. On the other hand, Ca +2 and EDTA (1 to 5 mM) did not significantly affect the enzyme activity. The main substrate for the enzyme was found to be starch but it could also hydrolyze raw starch, dextrin, γ γ γ γ-cyclodextrin and pullulan.
    AFRICAN JOURNAL OF BIOTECHNOLOGY 08/2011; 10(32):6082-6089.
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    ABSTRACT: Urinary tract infection (UTI) forms the largest single group of hospital-acquired infection and account for about 40–50% of the total nosocomial infections. Present study indicates that there is a high prevalence of multidrug resistant E. coli in UTI. Purpose of this study was to find out the occurrence of multidrug resistance among the Escherichia coli, which is the causative agent in more than 80% of urinary tract infection. In this study, 100 isolates were taken from UTI patients and their susceptibility pattern was studied. Minimum inhibitory concentration (MIC) of 11 antibiotics was studied using agar dilution method. The isolates were highly susceptible against imipenem i.e. 91% and were least susceptible to ampicillin and tetracycline. The resistance against ciprofloxacin was also very high i.e. 85%. ESBL production among these E.coli isolates was checked by combination disc method and about 54% isolates were found to be ESBL producers. So our result indicates the fact that the physicians should be aware of this increasing resistance among our local clinical isolates and should change their therapy regime accordingly.
    Australian Journal of Basic and Applied Sciences 01/2011; 7(1):39-43.
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    ABSTRACT: Chronic suppurative otitis media (CSOM) is a prevailing and notorious infection in developing countries causing serious local damage and threatening complications. Early and effective treatment based on the knowledge of causing micro organisms and their sensitivity results in good clinical recovery and prevents from damage and complications. Pseudomonas aeruginosa is most common pathogen causing CSOM in Pakistan. The objective of this study is to identify incidence of Pseudomonas aeruginosa involved in CSOM and sensitivity against commonly prescribed antibiotics. A total of 263 patients with unilateral or bilateral chronic suppurative otitis media (CSOM) were enrolled in the study at Department of ENT, Karachi Medical and Dental College & Abbasi Shaheed Hospital, Karachi, Pakistan from December 2004 to May 2006. Sterile cotton swabs were used to collect pus from discharging ears and plated on Blood agar and MacConkey agar for 24 to 48 hours. Antibiotics susceptibility was tested by Minimum inhibitory concentrations (MIC) method using Mueller-Hinton agar. Overall microbiology of 267 samples from 263 patients was studied including 4 of bilateral discharge. Polymicrobial growth was present in 8 samples. A total of 275 bacterial isolates were studied. Pseudomonas aeruginosa (40%) and Staphylococcus aureus (30.9%) were the most common bacterial agents found in CSOM. MIC was done for Pseudomonas aeruginosa only as it was the commonest pathogen found in CSOM. Sensitivity pattern of Pseudomonas aeruginosa showed that amikacin was active against 96% of isolates followed by ceftazidime 89%, ciprofloxacin 85%, gentamicin 81%, imipenem 76%, aztreonam 42% and ceftriaxone 21%. Pseudomonas aeruginosa was the most common bacteria isolated from chronic discharging ears followed by Staphylococcus aureus. Amikacin was found to be the most suitable drug followed by ceftazidime and ciprofloxacin for Pseudomonas aeruginosa. The resistance against ceftriaxone and aztreonam was found to be very high.
    Journal of Ayub Medical College, Abbottabad: JAMC 11/2008; 21(2):120-3.
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    ABSTRACT: Diabetic nephropathy is the leading cause of death that affects more than 40% of diabetic patients. Its metabolic derangements are frequently accompanied with electrolyte imbalances. This study was aimed to evaluate the electrolyte homeostasis during the progression of diabetic nephropathy in various stages of developing nephropathy. Patients admitted in diabetic wards of various hospitals of Karachi were selected and divided into 4 groups with 50 individuals each. Group I (healthy normotensive, non-diabetics with normal renal functions as control). Group II (diabetic patients with normal blood pressure and renal functions). Group III (diabetic hypertensive patients without renal disease). Group IV (diabetic nephropathy patients with nephropathy). Their fasting blood samples were drawn and analyzed for the estimations of intra erythrocyte and serum electrolytes and NA(+)-K(+)-ATPase activity. Group II patients showed a significant increase in intra erythrocyte sodium, serum potassium and calcium levels where as intra erythrocyte potassium, NA(+)-K(+)-ATPase, serum sodium and magnesium were significantly decreased as compared to control. Group III showed a significant rise in intra erythrocyte sodium levels but intra erythrocyte potassium, NA(+)-K(+)-ATPase, serum sodium, calcium and magnesium were significantly lowered as compared to control. Group IV revealed a significant increase in intra erythrocyte sodium and significant decrease in intra erythrocyte potassium, NA(+)-K(+)-ATPase, serum sodium, calcium and magnesium levels as compared to control. The results suggest the progressive trends in electrolyte abnormalities in diabetes mellitus leading to end stage renal disease along with the abnormality of their chief transport mechanism. It points towards the potentiality of electrolytes disturbances as indicators for the progression of diabetic nephropathy and also beneficial in prognosis and treatment of the disease.
    Pakistan journal of pharmaceutical sciences 05/2008; 21(2):172-9.
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