review on requirements for vitamin D fortification in Pakistan

Despite the fact that Pakistan is a country situated near equator and has plenty of sunshine, it still has major population deficient in vitamin D3. The majority of food opted by general public is not sufficient source of vitamin D. For the reason vitamin D fortification is a compulsory requirement for healthy living. Developed countries like United Kingdom and Canada have overall improved their population vitamin D status by fortifying foods with vitamin D. For applying a well maintained vitamin D food fortification strategy in Pakistan , it is necessary to review various methods , type of food and their fortification results implemented all over the world. This review investigates all such prospective approaches which could be used for the fortification of foods in Pakistan. Looking at the socio economic status the majority of Pakistani residents, vitamin D fortification of staple foods such as whole grains, flour and rice by spray methods might serve as viable approaches in targeting majority of Pakistani population to ingest and improve their vitamin D status

Microbiological load of ethylene oxide sterilized medical devices and its elimination by cobalt 60 source

Objective: To determine the residing microbial flora of ethylene oxide [EtO] sterilized medical devices and optimization of safe dose of gamma radiation [Cobalt 60 source] for the complete elimination of microbial load

Study Design: Experimental study

Place and Duration of Study: Department of Biotechnology, Lahore College for Women University, Lahore, Pakistan from September 2014 to June 2015

Methodology: Thirty-six samples of EtO sterilized medical devices of same batch of three different companies were collected for this study. Isolation and enumeration of microbes were done by using different selective and differential media. Gram staining and biochemically characterization by API 20 [Bio Merieux, France] kit was done for identification of the microorganisms. The medical devices having high microbial load were sent to Pakistan Radiation Services [PARAS] for gamma irradiations at 3 different selected doses [20 KGy, 25 KGy, and 30 KGy]

Results: Different types of Gram positive bacteria [Staphylococcus epidermidis, Staphylococcus aureus and Bacillus subtilis] were isolated from the EtO sterilized samples. Gram negative bacteria and fungi were not detected on these medical devices. Gamma irradiations results showed that 30 KGy was optimized dose for complete elimination of microbial flora on endotracheal, Nelaton, and tracheostomy tubes

Conclusion: Gamma radiations [Co 60 source] effectively decontaminate the microbial flora on the equipment previously sterilized by the ethylene oxide gas; and 30 KGy is the optimized dose for all these medical devices

Optimization of PCR for rapid detection of CTX-M gene in ESBL producing Klebsiella pneumoniae clinical isolates from Punjab, Pakistan

Aims: The study was carried out firstly, to evaluate the prevalence of extended spectrum beta lactamase (ESBL), multidrug resistant Klebsiella pneumoniae isolates from Punjab, Pakistan and secondly, to characterize the genotypes of their beta lactamase producing enzymes and optimization of PCR based method for rapid and authentic detection of antibiotic resistant gene. Methodology and results: Two hundred of K. pneumonia strains were isolated from different clinical samples. Blood and MacConkey Agar were used to isolate and identify bacterial microorganisms while Muller Hinton Agar was used to evaluate the antimicrobial susceptibility against different antibiotics as per CLSI 2012 guidelines. ESBL producing bacteria were screened by double disk synergy /combination disk test. PCR was optimized and performed for resistant gene (CTX-M). The results showed that most of the isolates were resistant to multiple antibiotic including cephalosporin, aztreonam, sulphamethoxazole/trimethoprim, ciprofloxacin, doxycyclin and were sensitive to imipenam and amikacin. Frequency of CTX-M gene in ESBL producing K. pneumoniae was 94%. Conclusion, significance and impact of study: Based on the finding of this study it is suggested that prevalence of CTX-M gene (95%) is very high among ESBL producing isolates. Therefore, PCR based method may help clinicians for rapid detection and treatment of patients by choosing right medication against the resistant bacteria as early as possible.

Evaluation of antimicrobial activity of extracts of in vivo and in vitro grown Vinca rosea L. [Catharanthus roseus] against pathogens

The antimicrobial activity of Vinca rosea was evaluated against pathogenic bacterial strains [Bacillus subtilis, B. licheniformis and Azotobacter sp.] and fungal strains [Asprgillus niger, Alternaria solani and Rhizopus oryzae] using agar well diffusion method. Methanolic extracts of in vivo leaf, in vitro leaf, in vitro calluses of leaf, nodal and fruit explants were used and exhibited antimicrobial activity as indicated by minimum inhibitory concentration [MIC]. In vitro extracts showed better results as compared to the in vivo extracts for both the antibacterial as well as the antifungal activity. Among all the extracts, maximum zone of inhibition [30.3 mm+/-0.58a] was formed by in vitro leaf callus extract concentration of 2.0mg/ml against B. licheniformis. Similarly in case of antifungal activity, maximum zone of inhibition [34.6mm +/-0.57a] was formed by in vitro leaf callus extract and MIC value is 6.0mg/ml against A. niger. Hence these results clearly depicts that V. rosea possess a great strength to fight against the microbial activity and can be used against various infections.

Significant association of serum creatinine with HbA1C in impaired glucose tolerant Pakistani subjects

The present study was conducted to assess the serum concentration of creatinine and determine its relationship with potential risk factors of diabetes in Impaired Glucose tolerance subjects. This cross sectional study was conducted on 100 IGT patients who attended Amin Hayat diabetic center in Lahore from January 2011- June 2011. Patients with age group 34-67 years, [both sexes] were included in the study. Different demographic parameters as age, BMI, WHR, B.P, personal history and socioeconomic status were recorded. Oral Glucose Tolerance Test was performed. The biochemical parameters including HbA1c, lipid profile, urea, uric acid, creatinine and bilirubin level were measured by chemistry analyzer. A strong correlation between creatinine and HbA1c was observed. The level of creatinine was also significantly associated with age in IGT subjects. Creatinine is non-significantly correlated with Cholesterol, LDL-Chol and TG while negatively significantly associated with BMI, fasting blood glucose and HDL-Chol. The present study concluded significant association of serum creatinine with HbA1c, BMI and HDL cholesterol

ABO blood group frequency in Ischemic heart disease patients in Pakistani population

To determine if there is any significant association between ABO blood groups and ischemic heart disease [IHD]. The study was performed at Punjab Institute of Cardiology [PIC], Lahore. Study duration was from January 2012 to September 2012. This study included 200 IHD patients and 230 control individuals. Self design questionnaire was used to collect information regarding risk factors. Standard agglutination test was performed to determine the blood groups. Data was analyzed on SPSS 16. The prevalence of blood groups in IHD group was 34% in blood group A, 29% in blood group B, 14% in blood group AB and 23% in blood group 0. In control group the distribution of B, A, AB and 0 blood groups were 34.4%, 20.9%, 12.6%, 32.2% respectively. Rh+ve factor was prevalent in 90.5% among IHD group and 92.6% in control subjects. The prevalence of IHD was more in males [63.5%] as compared to females [36.5%]. Mean age was 56.4 +/- 0.86 [yrs] and BMI was 26.4+/-0.33 [kg/m[2]]. The prevalence of hypertension was 58.5%, diabetes was 53%, family history of cardiac disease was 45%, 35.5% of patients were doing exercise regularly, 58.5% used ghee, and 58% were smokers. Subjects with blood group A had significantly [p< 0.05] higher risk of developing IHD as compare to other blood groups

Evaluation of antifungal and antioxidant potential of two medicinal plants:Aconitum heterophyllum and Polygonum bistorta

Objective:To focus on the evaluation of antimicrobial and antioxidant activity of two endangered medicinal plants Aconitum heterophyllum (A. heterophyllum) and Polygonum bistorta (P. bistorta). Materials: Plant extracts were obtained by using microwave assisted extraction method. The in vitro antifungal activity of A. heterophyllum and P. bistorta extracts were determined by measuring diameters of inhibitory zones of these extracts against Aspergillus niger and Alternalia solani. Results:Methanolic extract of A. heterophyllum showed significant (P≤0.05) antifungal activity against both the tested organisms. It was also observed that ethanolic extracts of P. bistorta also had good antifungal activity against the tested fungal strains as compared to the methanolic extracts. It showed significant antifungal activity (P≤0.05) against both the tested strains. Antioxidant activity of methanolic and ethanolic extracts of A. heterophyllum and P. bistorta were also measured using a radical scavenging method. Ascorbic acid was used as a standard. Conclusions:It was observed that A. heterophyllum and P. bistorta have significant antioxidant activity. Higher antioxidant activity was recorded in methanolic extract of A. heterophyllum as compared to its ethanolic extract. However, in case of P. bistorta ethanolic extract of the plant exhibited higher antioxidant potential than methanolic extracts. Hence both of these plants have significant antimicrobial as well as antioxidant potential.

Effect of stress on biofilm formation by icaA positive and negative strains of methicillin resistant Staphylococcus aureus

To determine the effect of physical and chemical stress factors e.g. antibiotics, NaCI, glucose, heat shock, cold shock and sonic waves on biofilm formation by icaA positive and negative strains of Methicillin resistant Staphylococcus [S.] aureus. Experimental study. Microbiological Analytical Centre, Pakistan Council of Scientific and Industrial Research [PCSIR] Laboratories Complex, Karachi, from January to December 2010. One strain of Staphylococcus aureus labelled as FA was isolated from a food sample and the other strain labelled as CL was a clinical strain. Biofilm assays were performed in brain-heart infusion [BHI] medium and in BHI supplemented with 7% NaCI, 5% glucose, or sub-inhibitory concentrations of Vancomycin, Oxacillin, Ampicillin, Tetracycline, Erythromycin, Rifampicin and Ciprofloxacin. Polymerase chain reaction [PCR] was used for screening of the icaA and mecA genes. The FA and CL were identified as MRSA carrying mecA gene. The strain FA showed biofilm formation without any treatment and was found to carry icaA gene contrary to CL, that does not contain this gene therefore, is unable to produce biofilm under normal conditions without any stress. The use of sub-lethal doses of cell wall active antibiotics, exposure to 7% NaCI, sonication, and heat shock were found to augment biofilm quantity in FA, an icaA positive strain and induce biofilm mode of growth in CL, an icaA negative strain. Anti-protein synthesis antibiotics did not show any effect on biofilm formation process in icaA positive or negative strains. There is a role of anti-cell wall factors i.e. sonication, heat shock, NaCI and antibiotics in the induction of biofilm mode of growth in MRSA and Methicillin sensitive S. aureus. The factors which partially damage bacterial cell wall, equally, induce biofilm formation in icaA positive or negative S. aureus

Associations between the Plasticity Region Genes of Helicobacter pylori and Gastroduodenal Diseases in a High-Prevalence Area

BACKGROUND/AIMS: Genes associated with the Helicobacter pylori (H. pylori) plasticity region may play a role in the pathogenesis of H. pylori. We compared the genes jhp0940, jhp0947, and jhp0986 in H. pylori isolates from patients with different gastroduodenal diseases and in different age groups. METHODS: The H. pylori hyperplasticity region genes jhp0940, jhp0947, and jhp0986 were studied by PCR. We also evaluated whether these genes were related to the cytotoxin-associated gene (cagA) and histology findings. RESULTS: Of the patient cohort, 71 (62%) were positive for jhp0940, 67 (59%) for jhp0947, 12 (10%) for jhp0986, and 69 (60%) for cagA. jhp0940 (n=18, 67%) and jhp0947 (n=23, 85%) were found more frequently in duodenal ulcer (DU) patients than in gastritis patients (n=14, 39%; p=0.029 and p<0.001, respectively). Gastric ulcer (GU) was more frequently associated with jhp0940 (17 patients, 77%; p=0.003) than with gastritis (14 patients, 39%). Gastric carcinoma (GC) was more strongly associated with both jhp0940 (22 patients, 76%; p=0.003) and jhp0947 (22 patients, 76%; p=0.003) than was gastritis (14 patients, 39%). jhp0947 was more frequently associated with chronic active inflammation (58 patients, 87%; p=0.009) than with chronic inflammation (9 patients, 13%). Multivariate analysis demonstrated that jhp0947 was associated with DU (odds ratio, 6.1; 95% confidence interval, 1.87-20). CONCLUSIONS: The genes jhp0947 and jhp0940 were identified in H. pylori isolates from patients with GC and DU, while jhp0940 was also isolated from patients with GU. jhp0947 was independently associated with DU.