Comparative analysis of dental pulp stem cells and stem cells from human exfoliated teeth in terms of growth kinetics, immunophenotype, self-renewal and multi lineage differentiation potential for future perspective of calcified tissue regeneration.

Background and Objectives: Owing to high proliferation rate, multipotency and self-renewal capability, dental pulp stem cells (DPSC) and stem cells from human exfoliated teeth (SHED) have become stem cell source of choice for cell based regenerative therapies. We aimed to compare DPSC and SHED as stem cell sources with a future use in regeneration of calcified tissue. Methods: Explant derived human DPSC (n=9) and SHED (n=1) were cryopreserved, thawed and expanded for analysis of population doubling time, colony forming unit assay and efficiency. A growth curve was plotted to determine population doubling time, while colony forming numbers and efficiency was determined at plating cell densities of 5.6, 11.1 and 22.2 / cm2. The isolated cells were characterized for the presence of stem cell markers by immunophenotyping and immunofluorescence staining, and tri-lineage differentiation. Statistical analysis was performed by Pearson correlation, Exponential regression and two way Anova with Tukey test at p<0.05. Results: DPSC and SHED exhibited spindle shaped fibroblast like morphology. SHED was found superior than DPSC in terms of proliferation and colony forming efficiency. Immunophenotypes showed that DPSC contain 62.6±26.3 %, 90.9±14.8% and 19.8±0.1%, while SHED contain 90.5%, 97.7% and 0.1% positive cells for CD90, CD73 and CD105. DPSC were strongly positive for vimentin, CD29, CD73, while reactivity was moderate to weak against CD44 and CD90. SHED expressed vimentin, CD29, CD105, CD90 and CD44. Both were negative for CD45. Upon induction, both cell types differentiated into bone, fat and cartilage like cells. Conclusion: Cultured DPSC and SHED were proliferative and exhibited self-renewal property. Both DPSC and SHED expressed stem cell markers and were able to differentiate into bone, fat and cartilage like cells. Thus, these could be a suitable stem cell sources for cell based regenerative therapies.

Prevalence of Nutritional Anaemia with Association of Body Mass Index among Karachi University students, Pakistan s.

OBJECTIVE: The main objective of the study was to evaluate the health of University of Karachi students (aged 19 to 21) by observing the correlation of haemoglobin level with underweight BMI. METHODS: In this study 151 students were enrolled from the Department of Biotechnology, University of Karachi, Sindh Pakistan. Blood samples were collected and Haemoglobin estimation was performed by Sahli's Haemoglobinometer. BMI of each student was calculated as per WHO criteria for South Asian. RESULTS: Out of 151 students, 60 (39.7%) students had anaemia, among which 41(27.1%) were showing grade 1 anaemia and 19(12.5%) were showing grade 2 anaemia. BMI results showed that 50 (33.1%) students were underweight whereas, 19(12.5%) students were overweight and 12( 7.9%) students were obese. Correlation showed that 34 (22.5%) students had anaemia with underweight BMI. CONCLUSIONS: This study concluded that anaemia with underweight BMI is more prevalent among females especially adolescent girls. This could be as they followed poor dietary habits and had some infections. This data evaluated the health status of University students on a small scale.

Isolation and culture of dental pulp stem cells from permanent and deciduous teeth.

OBJECTIVE: To isolate dental pulp mesenchymal stem cells (MSCs) from non-infected human permanent and deciduous teeth. METHODS: It was an in-vitro experimental study. Human teeth were collected from 13 apparently healthy subjects including nine adults and four children. After decoronation dental pulps were extirpated from teeth and cultured via explant method in a stem cell defined media. Data was analyzed by descriptive statistics. RESULTS: As above MSCs emerged exhibiting fibroblast-like morphology. In vitro culture was positive for 100% (9/9) and 75% (3/4) of the permanent and deciduous teeth respectively. First cell appeared from deciduous teeth pulp in 10±6.2 days while permanent teeth pulp took 12.4±3.7 days. Together, 26.6±3.6 and 24.5±3.5 days were required for permanent and deciduous tooth pulp stem cells to be ready for further assays. CONCLUSIONS: The protocol we developed is easy and consistent and can be used to generate reliable source of MScs for engineering of calcified and non-calcified tissue for regenerative medicine approaches.

Influence of different metals on the activation and inhibition of α-amylase from thermophilic Bacillus firmus KIBGE-IB28.

Thermophilic Bacillus firmus KIBGE-IB28 produced extracellular α-amylase at temperature 70°C. Enzyme was partially purified by ammonium sulfate precipitation with 42.80 fold purification and specific activity of 1889.6 U/mg. Effects of various metals on enzyme activity were determined and it was found that enzyme activity boosted significantly in presence of Ca(2+), K(2+), Ba(2+), Co(2+) and Ni(2+) whereas Zn(2+), Mg(2+), Na(2+) and Cu(2+) were found inhibitory at concentration 10mM.

Effect of exercise training program in post-CRET post-CABG patients with normal and subnormal ejection fraction (EF > 50% or < 50%) after coronary artery bypass grafting surgery.

The aim of the present study is to compare the effect of exercise training program in post-Cardiac Rehabilitation Exercise Training (CRET), post-CABG patients with normal & subnormal ejection fraction (EF >50% or <50%) who have undergoing coronary artery bypass grafting (CABG) surgery. The study was conducted on 100 cardiac patients of both sexes (age: 57-65 years) who after CABG surgery, were referred to the department of Physiotherapy and Rehabilitation between 2008 and 2010 at Liaquat National Hospital & Medical College, Karachi. The patients undertook exercise training program (using treadmill, Recumbent Bike), keeping in view the Borg's scale of perceived exertion, for 6 weeks. Heart Rate (HR) and Blood Pressure (BP) were measured & compared in post CABG Patients with EF (>50% or <50%) at the start and end of the exercise training program. Statistical formulae were applied to analyze the improvement in cardiac functional indicators. Exercise significantly restores the values of HR and BP (systolic) in post CABGT Patients with EF (>50% or <50%) from the baseline to the last session of the training program. There appeared significant improvement in cardiac function four to six weeks of treadmill exercise training program. After CABG all patients showed similar improvement in cardiac function with exercise training program. The exercise training program is beneficial for improving exercise capacity linked with recovery cardiac function in Pakistani CABG patients.

The relationship between sleep and cognitive functioning in adult people.

The purpose of the current study is to find out if subjective sleep complaints will have less cognitive functioning in older people (50 years and above). Sleep was assessed with the subscale Sleep Problems of the Symptoms Checklist-90 (Arrendell & Ettema 1986). Cognitive performance was measured with the Mini Mental Status Examination (Folstein, et al 1975) which is used as a dependent variable. Subjective complaints would be negatively associated with cognitive performance, since in elder people biological sleep is likely to be related with cognitive changes. A group of 12 people were given the task of collecting data through purposeful sampling techniques. Sample size of 120 participants was assessed. Each member of the group collected data from 10 subjects. Pearson Correlation Moment was applied for data analysis. In older persons the coefficient of falling asleep difficulty was -0.05 (p=0.33) and for waking up too early the coefficient was -0.13 (p=0.012) while for restless sleep coefficient was calculated as -0.09 (p=0.094). The assumption was verified that sleep problems negatively associated with cognitive functioning.

Enhanced production of maltase (α-glucosidase) from newly isolated strain of Bacillus licheniformis KIBGE-IB4.

Maltase (alpha-glucosidase) hydrolyzes α-(1→4) glucosidic bond of maltose into two glucose molecules. It is widely used in various foods, beverages and also in textile and biofuel industries. During current study, various physicochemical parameters for maltase production from newly isolated strain of Bacillus licheniformis KIBGE-IB4 were optimized using one-factor-at-a-time methodology. It was found that Bacillus licheniformis KIBGE-IB4 produced maximum maltase at 37°C, pH-7.0 after 48 hours using wheat starch (2.5%) as carbon source along with peptone (1.0%), yeast extract (0.2%) and meat extract (0.4%) as nitrogen sources in fermentation medium. It is concluded that the optimization of various medium ingredients and conditions increases maltase production upto 6.74 fold from B. licheniformis KIBGE-IB4 as compared to previously reported media and this strain could be used for the commercial production of maltase.

Immobilization of pectin degrading enzyme from Bacillus licheniformis KIBGE IB-21 using agar-agar as a support.

Pectinase from Bacillus licheniformis KIBGE IB-21 was immobilized in agar-agar matrix using entrapment technique. Effect of different concentrations of agar-agar on pectinase immobilization was investigated and it was found that maximum immobilization was achieved at 3.0% agar-agar with 80% enzyme activity. After immobilization, the optimum temperature of enzyme increased from 45 to 50 °C and reaction time from 5 to 10 minutes as compared to free enzyme. Due to the limited diffusion of high molecular weight substrate, K(m) of immobilized enzyme slightly increased from 1.017 to 1.055 mg ml(-1), while Vmax decreased from 23,800 to 19,392 µM min(-1) as compared to free enzyme. After 120 h entrapped pectinase retained their activity up to 82% and 71% at 30 °C and 40 °C, respectively. The entrapped pectinase showed activity until 10th cycle and maintain 69.21% activity even after third cycle.

Dextranase: hyper production of dextran degrading enzyme from newly isolated strain of Bacillus licheniformis.

Dextranase, 6-alpha-D-glucan 6-glucanohydrolase catalyzes the degradation of dextran (polymer of D-glucose) in to low molecular weight fractions. Dextranolytic bacterial strains were isolated from various natural sources and plate assay methods were developed for screening of highest extracellular dextranase producing isolate. Bacillus licheniformis, identified on the basis of taxonomic characterization was subjected to UV radiation and highest enzyme producing mutant obtained led to 7 times more dextranase production than wild. Optimization of major physico-chemical parameters affecting enzyme production; including medium composition, pH, cultivation time and temperature revealed that maximum enzyme production was obtained in a self designed medium (pH 6.0) containing 1% Dextran 5000 Da, after 24 h culture incubation at 37 °C. Dextranase reported in this study is of great commercial importance as it is strictly inducible in nature and B. licheniformis being non-pathogenic removes the safety concerns associated with production of dextran fractions for clinical and pharmaceutical usage.