Glutathione s-transferase M1 and T1 gene deletions and susceptibility to acute lymphoblastic leukemia [ALL] in adults

Objective: Biotransformation of xenobiotics is critical for their metabolism and removal from the body which is carried out by xenobiotic metabolizing enzymes. Individuals carrying variants of genes that encode these enzymes have an altered ability to metabolize xenobiotics which may lead to an increased risk of acute lymphoblastic leukemia. The current study aimed to investigate the impact of GSTM1 and GSTT1 gene deletions in causing predisposition to adult ALL

Methods: The current case-control study involved 62 adult ALL patients and 62 age and gender matched healthy controls. Whole blood samples processed with standard phenol chloroform protocol for DNA isolation were genotyped using multiplex PCR approach for simultaneous identification of GSTM1 and GSTT1 deletions. The genotype frequency obtained for patients was compared to controls using odds ratio and chi-square

Results: The null genotype frequency of GSTM1 and GSTT1 in a group of adult ALL patients from Pakistan were 47% and 11% respectively. Deletion of GSTM1 and GSTT1 did not show statistically significant association with adult ALL [p=0.86 and p=0.35 respectively]. The combined GSTM1/GSTT1 deletion was observed in 2% patients and was not significantly associated with ALL in adults [p=0.85]

Conclusions: The results reveal that homozygous null polymorphism of GSTM1 and GSTT1 genes does not influence ALL susceptibility among adult patients. Cancer susceptibility associated with GST polymorphism varies with ethnic and geographic differences. Therefore, further investigation on different populations is needed to understand the role of these genetic variations in modifying adult ALL risk

Association and implications in triple negative and triple positive breast cancer: relationship with sociodemographic and reproductive factors in pakistan

Background and Objectives: Triple negative and triple positive breast cancer have adverse effects than other types of breast cancer. However, triple negative has poor prognosis with short survival as compared with triple positive breast cancer. Good prognosis is one of the key factors for successful treatment trial. This study aimed to find out the association of sociodemographic and reproductive features like parity, menopause, number of child bearing as risk factors in the development and prognosis of triple negative and triple positive breast cancer

Methods: This study is a part of an ongoing project which is being conducted in Karachi from 2013 to 2020. Informed consent from triple negative breast cancer [n=134] and triple positive breast cancer [n=87] patients were taken prior to their recruitment into the study. Demographic, anthropometric, reproductive and disease history of patients were recorded. Means, frequency distribution, data classification and association analyses were done by SPSS version 17.0

Results: Statistical analyses revealed that delayed first child bearing age and lower number of children are associated with the development of triple negative breast cancer. However, no significant effect of these parameters has been observed on the outcomes of triple positive breast cancer

Conclusions: Reproductive factors have more pathological implications than sociodemographic factors in both triple positive and triple negative breast cancer development. These findings might prove to be beneficial for effective and better breast cancer management

study on Catalase Gene Promoter Polymorphism-21 A/T [rs7943316] in Healthy Pakistani population

Background and Objective: Catalase [CAT] is an important endogenous antioxidant enzyme that detoxifies H[2]O[2] into water and oxygen, consequently limiting the deleterious effects of reactive oxygen species. It has suggested that CAT -21A/T [rs7943316] OMIM: 115500 gene promoter polymorphism is predominantly associated with different human disorders such as hypertension, cancers, diabetes, nephropathy, and other diseases accompanied by oxidative stress. This study was designed to investigate the prevalence of mutant T allele frequency in healthy individuals

Methods: The study group consisted of 110 healthy individuals were enrolled from Baqai Institute of Diabetology and Endocrinology [BIDE], Karachi, Pakistan, during the period of April 2010 to May 2013. DNA was isolated from leukocytes. Genotyping of CAT -21A/T [rs7943316] gene promoter polymorphism was carried out using thermal cycler followed by RFLP. Blast N analysis was performed for the confirmation of gene sequences

Results: In CAT -21A/T [rs7943316] gene promoter polymorphism, wild type genotype [AA] was observed in 18.26% and alterered genotype [AT/TT] found in 81.74% cases

Conclusions: Data demonstrates that frequency and distribution of mutant T allele was more prevalent as compared to wild type A allele in the study group

Cheminformatics-based anticoagulant study of traditionally used medicinal plants

Background: Medicinal plants, as a complementary medicine, have been used to treat various diseases since ancient times. These plants have numerous beneficial applications and are the source of certain conventional drugs. In diseases such as stroke and ischemia, which are caused by several factors, abnormal coagulation is an important causative factor. Accordingly, novel and effective therapies such as herbal remedies should be practiced to prevent such lethal diseases

Methods: Using the available databases such as Google Scholar and PubMed, the previously reported anticoagulant compounds and plants possessing anticoagulant activity were identified and collected in two separate lists. Next, the fast and cost-effective cheminformatics methods incorporated in PubChem were applied to detect some compounds similar to reported anticoagulants. Subsequently, 15 native medical plants of Iran containing the potential anticoagulants were selected. The selected plants were purchased and chopped, and the potential compounds were extracted by ethanol. Then three concentrations of extracts [1, 10, and 100 micro g per ml] were made. Finally, anticoagulant effect of the selected plants was evaluated by in vitro prothrombin time and activated partial thromboplastin time coagulation tests

Results: Among the 15 selected medicinal plants, three plants, including Terminalia bellirica [P=0.0019], Astragalus arbusculinus [P=0.0021], and Origanum vulgare [P=0.0014] showed a more promising anticoagulant effect in comparison to the control

Conclusion: The anticoagulant activity was identified for the first time in these three plants. Further in vivo study and mechanism of action assay are required to be performed on these three plants, which could be suitable candidates for use as natural anticoagulant medicines

Association of single Nucleotide Missence Polymorphism Val109Asp of Omentin-1 gene and coronary artery disease in Pakistani population: Multicenter study

Background and Objective: Coronary artery disease [CAD] is a most important cause of morbidity and mortality worldwide as well as in Pakistan. Recent studies have shown that the combination of obesity, insulin resistance and fluctuation in circulating adipocytokines levels is associated with the pathogenesis of coronary artery disease. Omentin-1 is recently found adipocytokine that is highly expressed in visceral adipose tissue. It has anti- inflammatory properties and is negatively correlated with ischemic heart disease. Therefore, this study was designed to investigate the relationship between omentin-1 Val109Asp polymorphism and CAD in Pakistani population

Methods: A total of 350 subjects were included in the study. Two hundred fifty were diagnosed witd coronary artery disease while 100 served as healthy controls. PCR-RFLP was performed at Dr. AQ. Khan Institute of Biotechnology [KIBGE] to analyze Val109Asp polymorphism. In this, case control study SPSS software version 16 [Chicago, IL, USA] was used for data analysis. Continuous variables and categorical variables were presented as mean+/-SD or in percentage. Independent sample test and chi-square test was performed to compare the differences in means between cases and controls. Genotype distribution was analyzed by chi-square test and results were presented as percentage and frequency. MultivariWe regression analysis indicated that Val109Asp SNP might be an independent risk factor for CAD susceptibility after adjustment for some well- known CAD risk factors including age, gender, body mass index, smoking, hypertension, diabetes mellitus and lipid abnormalities. There was estimation of odd ratios [OR] and 951 confidence intervals [CIs] to determine the correlation between genotypes and the risk of CAD. [p> 0.05], Genotype frequencies were compared by Chi-square test

Results: There was prevalence of Omentin-1 Val109Asp polymorphism in both case and control groups, However, Val/Asp [heterozygous mutant] genotype was detected more frequently in patients with CAD, OR[95%]=1.921; Cl=1.173-3.1469 in comparison of Asp/Asp and Val/Val genotypes

Conclusion: Individuals having Val/Asp heterozygous gemotype of omentin-1 gene polymorphism are at more risk of developing CAD in Pakistani population, further studies are required in different populations and ethnicities to confirm our findings

Fresh and aged human lymphocyte metaphase slides are equally usable for GTG banding

The identification of chromosomes for routine cytogenetic analysis is based on quality of metaphases and good banding pattern

Fresh slides of human lymphocytes have been shown to produce good bands for the identification of chromosomes morphology. G-bands by Trypsin using Giemsa [GTG] banding of aged slides is generally considered hard to get desired band pattern of chromosomes persistently. The current study is focused on GTG banding of aged slides. A total of 340 subjects including 290 primary infertile and 50 fertile were selected. The blood samples were drawn aseptically for cytogenetic analysis

Lymphocytes were cultured and GTG banding was done on 1440 glass slides

Giemsa trypsin banding of aged slides were done by adjusting average trypsin time for each month according to the slide age and metaphase concentration

Correlation analyses showed a significant and positive correlation between slide ageing and trypsin pre-treatment time. The results of this study suggest that, the fresh and aged human lymphocyte metaphases are equally usable for GTG banding

Serum and salivary minerals in dental caries

Dental caries is a multifactor disease, affecting people of all ages. Inorganic mineral of serum and saliva can also have protective role in dental caries. This study was carried out to evaluate and compare the possible role of salivary and serum factors like pH, adequate level of calcium, phosphate and fluoride in dental caries. A Total of 100 subjects aged 10-40 were selected. Decayed, missed and filled teeth [DMFT] were used as indices for scoring the dental caries and were distributed or divided into 4 groups on the basis of DMFT indices as 4-8 [Group I], 9-16 [Group II], 17-24 [Group III] and more than 25 [Group IV], while the control subjects had DMFT index equal to or less than 3 BH, calcium phosphate, fluoride and lactic acid were analyzed in saliva and serum. Patients of dental caries showed significantly low levels of calcium, phosphate, fluoride [P<0.001] and significantly high level of lactic acid [P<0.001] in all the groups as compared to control subjects. Prominent significant changes were observed in different groups. The salivary and serum pH, calcium, fluoride, phosphate and lactic levels were found to be significantly changed among the patients having dental caries. It can be concluded from the findings of present study that the adequate levels of calcium, phosphate and fluoride in saliva as well as serum are responsible for the significant deposition of these minerals in plaque which greatly reduces the developmental caries in the adjacent enamel

role of bezafibrate in lowering risk factors for cardiovascular disease in diabetic patients

A number of hypolipidaemic drugs have been shown to lower the raised plasma fibrinogen concentrations indicating that lipid and haemostatic system may act synergistically in pathogenesis of cardiovascular diseases. The current study examines the possible role of lipid lowering drug bezafibrate to induce change in blood coagulation system and the architecture of fibrin network in addition to its known effects on lipid profile in diabetic patients. Methods based on turbidity for measurement of mass length ratio of fibrin fibers, permeability of the networks and their tensile strength have been used to assess the alterations induced by lipid lowering agent. The study group consisted of 38 patients [18 male and 20 female; age 40 - 65 years]. The patients were categorized into two groups. Group one included 20 diabetic patients, selected from Liaquat National Hospital, Karachi, with normal lipid profile as control [9 male, age 51.88 +/- 1.93 years; 11 female, age 48.36 +/- 2.44 years]. Second group included 18 diabetic patients, selected from National Institute of Cardiovascular Disease, Karachi, with altered lipid profile i.e. hypertriglyceridaemia and/or hypercholesterolemia [9 male, age 47.33 + 3.08 years; 9 female, age 48.66 +/- 3.99 years]. Blood samples were collected after 12-14 hours fasting at the start of the study and after administration of Bezafibrate [200 mg TDS] for 4 weeks. The samples were analyzed for blood parameters like cholesterol, low-density lipoprotein-cholesterol [LDL-C], high-density lipoprotein-cholesterol [HDL-C], triacylglycerol [TG],-and fibrin network characteristics. In addition to the changes in lipid profile, significant alterations were observed in the characteristics of fibrin networks by the drug. Amongst the changes induced are the properties of networks that make the networks more lysable by fibrinolytic agents. Clinical and pharmacological implications of these findings need further investigations

Serum leptin and pituitary thyroid axis in hyperthyroid male patients

To see the correlation of serum leptin with pituitary-thyroid axis in hyperthyroid male patients and in euthyroid lean and obese control subjects. Four groups of males were included in the study, they comprised of 22 newly diagnosed, untreated hyperthyroid patients, 24 age, gender, BMI matched euthyroid lean subjects, 27 euthyroid obese subjects. Clinically and biochemically confirmed hyperthyroid patients were selected from the thyroid OPD of Atomic Energy Medical Centre, JPMC, Karachi. Healthy, age, gender, matched euthyroid [lean and obese] controls were selected from general population after checking their thyroid profile and clinical examination. Patients and controls suffering from diabetes or other endocrinal diseases and /or taking treatment for hyperthyroidism or steroids were excluded from the study. Ten ml of blood was drawn after an over night fast from each patient and control. Serum leptin was measured by EL1SA and FT4, FT3 and TSH were measured by radioimmunoassay [RIA], fasting blood sugar [FBS], triglycerides [TG], total cholesterol [TC], high density lipoprotein cholesterol [HDL-C] were checked by kit methods. Low density lipoprotein cholesterol [LDL-C] was calculated by the Friedwald's formula. The mean +/- SEM values of leptin in male hyperthyroid patients wasl.71 +/- 0.45 ng/ml, while in lean and obese control group it was 8.27 +/- 1.91 and 21.34 +/- 3.4 ng/ml respectively. In hyperthyroid patients, significantly low levels of serum leptin were found as compared to age, gender, BMI matched lean [p<0.005] and obese control group [p<0.005]. Obese control group had significantly higher values of leptin as compared to lean group [p<0.05]. No significant correlation of leptin was found with T4, T3, and TSH in hyperthyroid patients and lean and obese control subjects. Triglycerides [TG, p<0.005], total cholesterol [TC, p<0.005] and low dentistry lipoprotein cholesterol [LDL-C, p<0.05] were significantly lower in hyperthyroid patients as compared to control groups. Serum leptin level significantly correlated with the BMI [p<0.005], in hyperthyroid patients and euthyroid lean and obese control groups. There appears no relationship between serum leptin and pituitary thyroid axis in dysthyroid patients as well as in euthyroid subjects but it is strongly related with BMI in all subjects