Bromelain: Methods of Extraction, Purification and Therapeutic Applications

Bromelain is a concoction of sulfhydryl proteolytic enzymes. Depending upon the site of extraction it can be regarded as either stem bromelain (SBM) (EC 3.4.22.32) or fruit bromelain (FBM) (EC 3.4.22.33). Bromelain remain enzymatic active over a broad spectrumand endure a range of pH (5.5 to 8.0) and temperature (35.5 to 71 ºC). It is one of the extensively investigated proteolytic enzyme owing to its astonishing applications in various industries. This necessitated employing a strategy that result in highest purified bromelain in less steps and lowest cost. Use of modernistic approach such as membrane filtration, reverse micellar systems, aqueous two phase extraction and chromatographic techniques have shown promise in this regard. Besides its industrial applications, bromelain has been widely utilized as a potential phytomedical compound. Some of its reported actions include inhibition of platelet aggregation, anti-edematous, anti-thrombotic, anti-inflammatory, modulation of cytokines and immunity, skin debridement and fibrinolytic activity. It also assist digestion, enhance absorption of other drugs and is a potential postoperatively agent that promote wound healing and reduce postsurgical discomfort and swelling.

Vivax malaria; frequency, severity of thrombocytopenia and variation in red cell distribution width [RDW]

To determine the frequency and severity of thrombocytopenia and to evaluate the variation of red cell distribution width [RDW] in patients with acute vivax malaria. Cross sectional descriptive case series study. Six months. Liaquat University Hospital. All the patients with acute vivax malaria were evaluated for thrombocytopenia its severity and RDW. The data was analyzed in SPSS 10 and the frequency and percentage was calculated. Total 126 patients with acute vivax malaria were recruited, of which 88 were males and 38 were females. The mean age +/- SD for male and female subjects was 44.76 +/- 6.83 and 40.83 +/- 7.42. The common features observed were fever 31%, rigor 15% and combined features 14% [p=0.05]. The thrombocytopenia was identified in 86 patients of which 65 were males and 21 were females [p=0.04]. The increased RDW was observed in 75 patients of which 57 were males and 18 were females [p=0.05]. The bleeding was detected in 56 and dyspnea and heart failure was found in 30 thrombocytopenic subjects [p=0.02]. Regarding outcome 82 subjects were recovered while the 04 patients were expired [p=0.05]. Regarding severity of thrombocytopenia majority 48.8% were in moderate category with male predominance [p=0.02]. The thrombocytopenia and increased RDW was observed in patients with acute vivax malaria

Hypoglycemia; patients with liver cirrhosis

To determine the frequency and severity of hypoglycemia in patients with liver cirrhosis. Cross sectional case series study. Six months. Liaquat University Hospital Hyderabad. All the patients of liver cirrhosis, of >12 years of age and of either gender were evaluated for hypoglycemia by assessing the glycemic status through random or fasting blood glucose level. The severity of liver cirrhosis was identified according to the Child-Pugh classification whereas the severity of hypoglycemia was grouped in mild, moderate and severe categories. The data was entered and saved in SPSS and frequency and percentage was calculated for hypoglycemia in patients with liver cirrhosis. The stratification was done for age, gender, hypoglycemia and severity of the disease and hypoglycemia. The chi-square test was applied between categorical variables at 95% confidence interval and p -value

Metabolic syndrome; gamma glutamyltransferase in patients

To determine the role of gamma glutamyltransferase as a biochemical marker for the diagnosis of metabolic syndrome. Cross sectional descriptive study. One year. Department of Medicine, Liaquat University Hospital Hyderabad / Jamshoro. All the patients with metabolic syndrome visited at OPD / admitted in the ward were further evaluated for serum gamma-glutamyltransferase level. The data was analyzed in SPSS 16 and the frequency and percentage was calculated. During one year study period, total one hundred patients [23 males and 77 were females] with metabolic syndrome were recruited and study for gamma glutamyl transferase level. The mean age +/-SD for overall population was 56.84+/-6.52 whereas it was 48.92+/-5.82 and 58.61+/-7.73 in male and female population respectively. The mean +/- SD of systolic and diastolic blood pressure [mmHg], triglycerides [mg/dl], high density lipoprotein pressure [mg/dl] and fasting blood sugar [mg/dl] in overall population was 161.20 +/- 16.74 and 95.60 +/- 8.34, 176.38 +/- 11.93, 29.44 +/- 2.90 and 108.42+/- 6.25. The mean gamma glutamyl transferase level in overall population was 86.75+/-7.74 while it was 84.83+/-5.32 and 89.52+/-6.84 in male and female population respectively. The gamma-glutamyltransferase was raised in 75 patients of which 13 were males and 62 were females [p=0.02] and majority of patients were 50-59 year age group [p <0.01]. It is concluded that GGT is a good diagnostic tool in metabolic syndrome with statistical significant results

Liver cirrhosis; frequency and severity of hyponatremia in patients

Liver cirrhosis results from prolonged, widespread but patchy hepato-cellular necrosis due to various reasons. To determine the frequency and severity of hyponatremia in patients with liver cirrhosis. Descriptive case series study. Six months. Liaquat University Hospital Hyderabad. The cirrhotic subjects were assessed for hyponatremia and its severity. The data was analyzed in SPSS 16 and the frequency and percentage was calculated for hyponatremia and statistically p -value

Liver cirrhosis; frequency and severity of thrombocytopenia in patients

To determine the frequency and severity of thrombocytopenia in patients with liver cirrhosis. Cross sectional study. 01-03-2013 to 31-08- 2013. Liaquat University Hospital, Hyderabad. The cirrhotic patients were assessed for thrombocytopenia and its severity. The data was analyzed in SPSS version 11.00 and frequency and percentage was computed. The chi-square test was applied and p -value

Isolation and Identification of Glucose Oxidase Hyper Producing Strain of Aspergillus niger.

Aims: Glucose oxidase is an enzyme with large scale applications in various industries. It is also used in several diagnostic kits which makes it medically important as well. Our aim was to isolate indigenous glucose oxidase hyper producing strain of Aspergillus niger from different soil samples of Punjab, Pakistan. Study Design: An experimental study. Place and Duration of Study: Institute of Industrial Biotechnology, GC University, Lahore from March 2011 to July 2012. Methodology: Two hundred and seventy nine fungal strains were isolated from soil of different localities of Punjab. Isolates were screened for glucose oxidase production using submerged fermentation. Glucose oxidase hyper producer isolate was identified using morphological and molecular techniques i.e. 18S rDNA. DNA was isolated and amplified using PCR. Gene sequencing was done and homology analysis was studied. Rate of glucose oxidase production was also analysed. Results: Glucose oxidase hyper producing isolate was identified as A. niger A247 strain. This strain gave best reproducible results (145.22 ±0.034 U/g of cell mass) after 72 hrs of fermentation at 30ºC and at a medium pH of 7.2. Conclusion: Our results indicate the natural ability of A. niger to produce Glucose oxidase in large quantity instead of using genetic manipulation techniques.

Fibrinoid necrosis and hyalinization observed in normal, diabetic and hypertensive placentae

To determine the fibrinoid necrosis and hyalinization extent in placenta observed in normal, diabetic and hypertensive pregnancies. Comparative cross-sectional study. Institute of Basic Medical Sciences, Dow University of Health Sciences, Karachi, from 2008-2010. One hundred and fifty placentae were divided in three groups on the basis of their histories and clinical examination. Group A [control], Group B [Diabetic] and Group C [Hypertensive], each consisted of 50 samples. The samples were transferred to Dow Diagnostic Reference and Research Laboratory for histopathology and gross examination. The tissue samples were taken from different sites, processed and routine staining done. The slides were then examined under light microscope for hyalinization and fibrinoid necrosis. The data was analyzed by applying ANOVA and post-hoc Tukey at 95% confidence interval. Mean +/- standard deviations [SD] were computed. The mean number of hyalinized villi in control group was 0.54 +/- 0.908, 1.18 +/- 1.9540 in the diabetic group and 2.14 +/- 1.863 in the hypertensive group. The difference in their average turned out to be statistically significant [p-value < 0.001]. Mean number of villi having fibrinoid necrosis was statistically significant in both the diabetic and hypertensive groups as compared to the control group i.e. 13.98 vs. 4.02 and 10.08 vs. 4.02 respectively [p-value < 0.001]. There was significantly greater fibrinoid necrosis and hyalinization in placentae from mothers having diabetes and hypertension. The fibrinoid necrosis was seen more in diabetic group as compared to hypertensive and control, while hyalinization was observed more frequently in hypertensive group as compared to the other groups. Placental changes as seen in examination of delivered placentae will be helpful in preventing the adverse effects in successive pregnancies

Malaria morbidity in Sindh and the plasmodium species distribution

To determine the prevalence of Malarial parasites and species responsible in Sindh Province of Pakistan. It is a cross sectional descriptive study conducted in all Districts of Sindh during January 2002 to December 2006. It was based on surveillance data of Malaria control program Sindh in which blood smears from febrile cases in all age group of male and female were seen by facility microscopist in all districts. Senior microscopist rechecked all positive slides. All slides data of different districts of Sindh was reported on monthly basis to provisional centre at Hyderabad for compiling and analysis. During 2002-2006, out of 5.84 million slides, those with positive malarial parasites were approx. 0.16 million giving an average blood examination rate [BER] 4.46, slide positivity rate [SPR] 2.94, Falciparum ratio[FR] was 41%, annual parasite incidences [API] was 1.36. More cases were seen in post monsoon season. All basic indicators of Malaria disease are higher than optimum level. Estimated burden can be much higher, Roll Back Malaria goals are lagging far behind to reduce the disease burden. As such integrated approach of diagnosis, treatment and prevention is required