ABSTRACT
High prevalence of premature coronary heart disease in Pakistanis compared to other populations points towards the genetic predisposition of this population to develop this disease. Since no investigations have been carried out in Pakistan to study the relationship of polymorphisms in genes involved in homocysteine cycle, the objective of the present study was to find out if there is any association of methylenetetrahydrofolate reductase [MTHFR] C677T, A1298C; methionine synthase [MS] A2756G; cystathionine-beta-synthase [CBS] 844ins68, G919A polymorphisms with premature acute myocardial infarction [AMI] in a population of Pakistani patients with this disease. In a cross-sectional study, DNA samples of 143 AMI patients [age <45 years] and 153 healthy controls were genotyped for the above mentioned polymorphisms using PCR-RFLP methods. Plasma/serum samples of both patients and healthy controls were screened for homocysteine, folate and vitamin B12. One way ANOVA and chi-squared test were used for analysis of data. Mean plasma homocysteine levels in premature AMI patients and healthy controls were found to be 23 +/- 17.2 and 23 +/- 13.4 micro mol/l, respectively which are higher than the upper normal limit of this biomarker [15micro mol/l]. MTHFR 677 CT genotype in healthy controls and MTHFR 677 TT genotype in AMI patients were found to have significantly increased levels of plasma homocysteine [p value <0.05], while all other polymorphisms did not show any significant difference in mean levels of homocysteine between AMI patients and healthy controls. Moreover, no association was observed between MTHFR C677T, A1298C; MS A2756C; CBS844ins68 polymorphisms and premature AMI in this population. This indicates that common polymorphisms in MTHFR, MS and CBS genes have no role in premature AMI in Pakistani population
ABSTRACT
To investigate the role of dietary intake in the development of premature acute myocardial infarction [AMI] in a hospital-based Pakistani population in Karachi. In a case control study, 203 consecutive patients [146 males and 57 females] with their first AMI and age below 45 years were enrolled with informed consent. Similarly, 205 gender and age matched [within 3 years] healthy adults were also included as controls. Dietary intake of both cases and controls was assessed by using a simple 14-item food frequency questionnaire. Using factor analysis, 3 major dietary patterns- prudent dietary pattern, combination dietary pattern and western dietary pattern were identified. Fasting plasma/serum of both cases and controls were analyzed for homocysteine, folate, vitamin B[12], blood Pb, ferritin, cholesterol, LDL-cholesterol, HDL-cholesterol and triglycerides. ANOVAand conditional logistic regression were used to predict the association of dietary patterns with AMI. Consumption of prudent diet, characterized by high consumption of legumes, vegetables, wheat, chicken and fruits, is protective against the risk of premature AMI. Moderate to high consumption of combination diet, characterized by high intake of eggs, fish, fruits, juices and coffee was associated with decreased risk of AMI. No association was observed between western diet, characterized by high intake of meat, fish and tea with milk and risk of AMI. Consumption of a prudent dietary pattern and a combination dietary pattern is protective against the risk of AMI in a Pakistani population
ABSTRACT
Methylenetetrahydrofolate reductase [MTHFR] gene polymorphisms have been reported to be associated with response to methotrexate [MTX] in certain populations of patients with rheumatoid arthritis [RA]. This study aims at investigating any relationship of two single nucleotide polymorphisms [SNPs] in MTHFR gene, C677T and A1298C with response to therapy with MTX in Pakistani RA patients. Allelic frequencies of the two polymorphisms [C677T and A1298C] were determined in 67 RA patients [9 males and 58 females; mean age 42.87 +/- 13.5 years] who had previously participated in a prospective clinical trial. Fifty-one patients had received MTX and were followed up for response up to 6 months. Genotyping of the two MTHFR polymorphisms was carried out using PCR-RFLP, while fasting concentration of plasma homocysteine was determined using a kit method. Twenty-eight patients were found to be "good responders", while twenty-three were [poor responders]. MTHFR 1298C and MTHFR 677T alleles' frequencies in [good responders] were not different from frequencies in [poor responders] [0.574 vs. 0.521; p=0.6 and 0.197 vs. 0.196; p=0.75, respectively]. Plasma homocysteine levels in female RA patients were significantly higher compared to general population in Karachi [13.1 +/- 6.7 micromol/1 vs. 11.4 +/- 5.3 micromol/1; p<0.00l]. MTHFR C677T and A1298C polymorphisms are not associated with response to MTX in a population of Pakistani RA patients
Subject(s)
Humans , Adult , Female , Male , Middle Aged , Methotrexate , Prospective Studies , Polymorphism, Genetic , Methylenetetrahydrofolate Reductase (NADPH2)ABSTRACT
Apolipoprotein E [Apo E] is a basic component of very low-density lipoprotein [VLDL] and high-density lipoprotein [HDL]. It plays an important role in the clearance of cholesterol from circulation, and thereby slows down the process of atherosclerosis. Its 3 isoforms, E2, E3 and E4 are encoded by 3 alleles, e2, e3 and e4, respectively. E3 is the most common isoform in most populations in the world, while E2 is the least frequent isoform. A meta-analysis of several studies conducted on Asian populations revealed that carriers of e4 allele had 42% higher risk of coronary heart disease [CHD], while e2 allele had no significant association with this disease. Epidemiological studies performed in this region showed that E3 was the most prevalent isoform and most likely to be involved in CHD in Asia. Studies conducted in India indicated that individuals with e3/4 and e4/4 genotypes had considerably higher prevalence of dyslipidemia as compared to other genotypes, and hypertensive patients had high frequency of e4 allele. However, several other studies failed to show a relationship between ApoE gene polymorphism and CHD. Inadequate statistical power, low allele frequency, CHD phenotypes could be some of the possible reasons for conflicting results of some of these studies. This area of research is wide open and epidemiological studies with large sample size would be required to ascertain the relationships between ApoE gene polymorphism and CHD
ABSTRACT
Low doses of granulocyte- colony stimulating factor [G-CSF] and granulocyte macrophage- colony stimulating factor [GM-CSF] have been shown to be beneficial in reducing duration of systemic antibiotic therapy and in-patient hospitalization by decreasing the period of neutropenia in cancer patients undergoing chemotherapy. Since the underlying mechanism is unclear, the aim of this study was to investigate whether the administration of G-CSF and GMCSF in two different doses [low dose and standard dose] would result into resolution of neutropenia with concomitant increase in multiple forms of dihydrofolate reductase [DHFR, a pivotal enzyme in the pathway of de novo DNA synthesis]. Thirty seven cancer patients [26 males and 11 females; age 14-73 years] having chemotherapy-induced neutropenia [absolute neutrophil counts <500/ micro l] were treated with colony stimulating factor [CSF] in the following manner: 11 received GM-CSF [7 received a dose 250 micro g/m[2] and 4 received a dose of 100 micro g/m[2]]; 26 received G-CSF [14 received a dose of 5 micro g/kg and 12 received a dose of 2.5 micro g/kg]. CSFs was given every day till the absolute neutrophil count was more than 1,000/ micro l. Ten ml blood was collected from each patient and analyzed for total leukocyte count [TLC] and active DHFR and immunoreactive nonfunctional form of DHFR [IRE] in the cytoplasm of blood leukocytes by using methotrexate binding assay and enzyme-linked immunosorbent assay [ELISA]. A significant increase [p<0.05] in concentrations of both active DHFR and IRE following stimulation with low as well as standard doses of CSFs was observed along with increase in the TLC. There was no significant difference in number of days to resolution of neutropenia at these two doses, indicating that even low doses of CSFs are clinically effective. Along with an increase in TLC, the levels of DHFR increased even at low doses of CSF suggesting that this might be one of the mechanisms for CSF-induced proliferation of leukocytes in neutropenic cancer patients
ABSTRACT
Trans fatty acids [TFA] are produced either by hydrogenation of unsaturated oils or by biohydrogenation in the stomach of ruminant animals. Vanaspati ghee and margarine have high contents of TFA. A number of studies have shown an association of TFA consumption and increased risk of cardiovascular disease [CVD]. This increased risk is because TFA increase the ratio of LDL cholesterol to HDL cholesterol. Food and Agriculture Organization of the United Nations and World Health Organization have come up with the recommendation that the contents of TFA in human dietary fat should be reduced to less than 4%. There is high prevalence of CVD in Pakistan. High consumption of vanaspati ghee which contains 14.2-34.3% of TFA could be one of the factors for this increased burden of CVD in Pakistan. Consumption of dietary fat low in TFA would be helpful in reducing the risk of CVD in South Asia. Denmark by banning the sale of food items with TFA has brought down the number of deaths due to coronary heart disease by nearly 50% over a period of 20 years. Public awareness about the adverse effects of TFA on human health would be extremely important. Media can play a very effective role in educating the masses and advocating the policy for the sale of only low TFA food items. Google and US National Library of Medicine, National Institute of Health were the sources of papers cited in this review article
ABSTRACT
Lead [Pb] pollution is a serious problem in the developing countries, especially in South Asia. No levels of blood Pb can be considered safe for human health. While hypertensive effects of Pb have been well studies, there are only a few studies on the relationship of Pb exposure to the development of cardiovascular disease [CVD] in the Asian developing countries. The underlying mechanism is still unclear; however, oxidative stress and hyperhomocysteinemia appear to be most plausible hypotheses for the development of atherosclerosis leading to CVD. Enforcement of regulations for control of Pb pollution, increased public awareness about the harmful effects of Pb, cessation of smoking, frequent use of fruit juices and avoidance from iron and folate deficiencies would be required to protect masses from the deleterious effects of Pb
ABSTRACT
The countries of South Asia have been afflicted with poverty and are under tremendous economic strain. The high prevalence of cardiovascular disease [CVD] due to urbanization and adoption of unhealthy life style is putting further stress on the economy of these countries. The projected cost of CVD in terms of lost GDP by 2015 could be 31 billion US dollars in Pakistan and 237 billion dollars in India if appropriate measures are not adopted to decrease the burden of these diseases. Major risk factors associated with CVD are fortunately modifiable such as tobacco use, alcohol use, hypertension, obesity, high cholesterol, high glucose, low intake of fruits and vegetables and physical inactivity. By adopting policies for control of tobacco use, alcohol use, easy availability of health-promoting foods, provision of opportunities for engaging in physical activity, control of pollution, dissemination of health promotion messages through media and school curricula and introduction of cost-effective screening programs the burden of CVD could be reduced in this region, thereby having a positive impact on the economy of South Asian countries
ABSTRACT
To find out the total antioxidant status [TAS] and its relationship to blood lead [Pb] in apparently healthy adults in a population in Karachi. In a cross-sectional study, a cohort of 132 healthy adults [67 having blood Pb levels below 7 micro g/dl and 65 having blood Pb levels above 11 micro g/dl] were randomly selected from 872 healthy adults [age 18-60 years] recruited from a low-income population in Karachi. TAS in serum was analyzed using spectrophotometric assay based kit, while blood Pb levels were determined using Atomic Absorption Spectrometric method. Mean serum TAS levels were significantly less in subjects with high blood Pb concentration compared to those with low blood Pb concentration [1.05 +/- 0.27 mmol/l vs 1.16 +/- 0.24 mmol/l; P=0.01]. In a correlation analysis adjusted for age and gender, TAS levels were found to be modestly associated with blood Pb [Pearson's r=-0.2; P=0.02]. Mean TAS levels were significantly decreased in apparently healthy adults with high blood Pb in a general population in Karachi. Exposure to Pb pollution could be compromising antioxidant ability of individuals in the study population
Subject(s)
Humans , Male , Female , Lead/blood , Adult , Cross-Sectional Studies , Spectrophotometry , Spectrophotometry, AtomicABSTRACT
To find out the prevalence of hyperhomocysteinemia, and deficiencies of folate, vitamin B6 and vitamin B12 in an urban population in Karachi, Pakistan. In a pre and post experimental study, eight hundred and seventy-two apparently healthy adults [aged 18-60 years; 355 males and 517 females] were recruited from a low-income urban locality in East of Karachi from February 2006 to March 2007. Fasting venous blood was obtained. Serum was analyzed for folate and vitamin B12. Plasma was analyzed for pyridoxal phosphate [PLP, coenzymic form of B6] and total homocysteine. A group of vitamin-deficient individuals [n=194] was given 3-week supplementation with folic acid [5mg/day], methycobalamin [0.5mg/day] and pyridoxine hydrochloride [vitamin B6, 50 mg/day]. After supplementation, serum/plasma levels of folate, vitamin B12, PLP and homocysteine were again determined. Prevalence of hyperhomocysteinemia [>15micromol/l] was 32%. Similarly percent values of folate deficiency [<3.5ng/ml], vitamin B6 deficiency [PLP<20 nmol/l] and vitamin B12 deficiency [<200pg/ml] in the study population were 27.5%, 33.7% and 9.74%, respectively. Hyperhomocysteinemia was associated with male sex, folate deficiency, vitamin B12 deficiency [OR [95%CI], 8.3[5.7-12.1]; 2.5[1.76-3.58]; 2.6[1.5-4.5], respectively]. A 3-week supplementation with folic acid, methycobalamin and pyridoxine hydrochloride in vitamin-deficient subjects decreased plasma homocysteine levels by 37%. High prevalence estimates of folate, vitamin B12, and vitamin B6 deficiencies appear to be the major determinants of hyperhomocysteinemia in a low income general population in Karachi
Subject(s)
Humans , Adolescent , Adult , Middle Aged , Male , Female , Folic Acid/blood , Folic Acid Deficiency/epidemiology , Vitamin B 6 Deficiency/epidemiology , Vitamin B 12 Deficiency/epidemiology , Urban Population , Cross-Sectional StudiesABSTRACT
Mild grade anemia in adult population is defined as a blood hemoglobin concentration between 10.00 to 12.9 g/dl in males or between 10.00 to 11.9 g/dl in non-pregnant females. Mildly anemic but apparently healthy subjects are commonly found in general population. Such individuals are at a greater risk of not being sufficiently investigated for the underlying disease and may end up with serious morbidity and health problems such as, diminished cardiovascular responses, compromised physical activity, poor cognitive function and growth retardation in children, and increased risk of falls and fractures and 5-year all cause mortality rates among elderly people. Nutritional deficiencies, chronic diseases, recurrent infections and aging are the major causes of mild anemia. The best approach to combat mild anemia in general population of a developing country would be through initiation of food fortification program, creating more awareness among people about the benefits of dietary diversification, use of fresh fruits and green leafy vegetables and adopting measures such as, frequent hand washing and use of boiled water for drinking to avoid common infections
Subject(s)
Humans , Male , Female , Anemia/mortality , PopulationABSTRACT
To determine the plasma/serum levels of homocysteine, and vitamins folate, B6 and B12, in Pakistani healthy adults. Cross-sectional study. The Aga Khan University, from October 2006 to April 2008. Fasting levels of plasma/serum folic acid, pyridoxal phosphate [PLP], vitamin B12 and homocysteine were determined in 290 apparently healthy hospital personnel from institutions in two cities of Pakistan. Spearman correlation test and linear regression analysis was conducted. There were 219 males and 71 females with mean age of 46 +/- 10.5 years and mean body mass index of 23.5 +/- 3.8. Mean plasma homocysteine levels in Pakistani normal adults were found to be 17.95 +/- 8.4 mol/l. Mean concentrations of plasma/serum folate, vitamin B12 and PLP were found to be 5 +/- 3.9 ng/ml, 522 +/- 296 pg/ml and 21.6 +/- 14 nmol/l, respectively. Serum/plasma levels of folate, vitamin B12 and PLP were negatively correlated with plasma homocysteine [rho coefficient=-0.367, p < 0.001; -0.173, p=0.004; -0.185, p=0.002, respectively]. Serum folate and plasma PLP levels were inversely related with plasma homocysteine, adjusted for gender, age, smoking and body mass index [p < 0.001 and p=0.003, respectively]. Percent deficiency values of folate, vitamin B6 and vitamin B12 were 39.7%, 52.8% and 6.6% respectively. The high levels of plasma homocysteine could indicate a reason for mass micronutrient supplementation to prevent the high incidence of cardiovascular disease observed in Pakistani population
Subject(s)
Humans , Male , Female , Folic Acid Deficiency , Vitamin B 6 Deficiency , Adult , Homocysteine , Cross-Sectional Studies , Dietary Supplements , Vitamin B 12 Deficiency , Pyridoxal Phosphate , Demography , Body Mass IndexABSTRACT
A relatively simple spectrophotometeric method has been developed for the determination of a tocopherol in human plasma. Method is modification of a previous micromethod base on oxidation of a tocopherol by ferric chloride. The complex of ferrous ions [generated in this reaction] with bathophenanthroline is determined spectrophotometrically at 536 nm. The absorbance of this colored complex is directly proportional to concentration of a tocopherol. The method involves extraction of a tocopherol from human plasma using nhexane which is relatively less toxic than xylene. The assay is sensitive enough to detect as little as 0.2 microg of vitamin. The recovery of a tocopherol from the plasma using n-hexane was in the range of 75% - 100%. The mean values of intra-assay and interassy coefficient of variation were found to be 5.3% and 13%, respectively. The assay was used to monitor a tocopherol levels in plasma samples of 81 normal healthy adults. Mean concentration of plasma a tocopherol in these normal healthy adults was found to be 9.45 +/- 2.64 microg/ml. Sixteen percent of adults had low levels of a tocopherol. The method is rapid, convenient, reproduciable and relatively less hazardous compared to methods using xylene for the extraction of vitamin E. It can be routinely used to analyze as many as 20 plasma samples in about 2 hours time
Subject(s)
Humans , alpha-Tocopherol/blood , Tocopherols/analysis , Tocopherols/blood , Clinical Laboratory TechniquesABSTRACT
Human serum paraoxonase is a high density lipoprotein [HDL]-bound enzyme exhibiting antiatherogenic properties. The aim of this study was to investigate any relationship between serum paraoxonase activity and serum levels of HDL-cholesterol in Pakistani patients with acute myocardial infarction [AMI] compared to normal healthy subjects and to examine possible association between serum paraoxonase activity and AMI in Pakistani population. In a case-control study, serum paraoxonase activity and serum levels of HDL-cholesterol and LDL-cholesterol were monitored in 164 Pakistani patients with AMI and 106 normal healthy adults matched for gender, BMI and age within 10 years. Mean serum concentration of HDL-cholesterol and mean serum paraoxonase activity in AMI patients were not significantly different from the corresponding values in normal healthy subjects. Mean serum paraoxonase activity value was significantly lower in normal healthy subjects with low HDL-cholesterol [serum levels<40mg/dl] compared to the value in those with normal levels of HDL-cholesterol [P=0.04]. In AMI patients, paraoxonase activity was lower in subjects with low HDL-cholesterol compared to those with normal levels of HDL-cholesterol, however, the decrease was not statistically significant. Correlation analyses of the data revealed a moderate association of paraoxonase activity with HDL-cholesterol [Pearson's r=0.225, P<0.01 for AMI patients and r=0.281, P<0.01 for normal healthy controls]. Seventy three percent of normal healthy subjects and 65% of AMI patients in this study had low HDL-cholesterol. Low serum paraoxonase activity and high prevalence of low HDL-cholesterol in Pakistani population could be contributing to the high rates of coronary heart disease in this population
ABSTRACT
To investigate the frequency and distribution of DRB1 and DQB1 alleles in Patients with rheumatoid arthritis [RA] and analyze the relationship between clinical response to methotrexate [MTX] and the HLA-DR and HLA-DQ genotypes in these patients. In this case-control study, the HLA-DRB1 and HLA-DQB1 polymorphism in 91 RA patients and 91 healthy controls was done using polymerase chain reaction and sequence specific primers. There was no statistical difference in frequencies of HLA-DRB1*03, DRB1*04, DRB1*07, DRB1*10, DRB1*11, DRB1*12, DRB1*13, DRB1*14, DRB1*15 and DRB1*16 genotypes between patients and controls. However, DRB1*01 was found to be significantly more common [p=0.015] in RA patients compared to controls. HLA-DRB1*15 was more common in patients [43.5%] compared to controls [30.8%] but results were not significant. HLA-DRB1*08 and DRB1*09 were present in negligible number in patients as well as controls while HLA-DRB1*12 was conspicuously absent in controls. Similarly, DQB1*06 was also significantly more common [p = 0.01] among the patients compared to healthy control subjects, while there was no statistical difference in the frequencies of DQB1*02, DQB1*03, DQB1*04 and DQB1*05 among the cases and the controls. RA susceptibility in most patients appeared to be associated with the HLA-DRB1*01/DQB1*06 genotype. Regarding association between HLA-DR or HLA-DQ genotype and clinical response to methotrexate [MTX], the data showed that with the exception of HLA-DRB1*03, there appears to be no association between the particular subtypes of HLA-DR and HLA-DQ. HLA-DRB1*03 was significantly more common among non-responders to MTX alluding to the possibility that another genes responsible for MTX metabolism, might be in linkage disequilibrium with HLA-DRB1*03 in the Pakistani population, thereby making such individuals non-responsive to MTX-therapy. RA susceptibility in most Pakistani patients is associated with the HLA-DRB1*01/DQB1*06 genotype. HLA-DRB1*03 was found to be significantly more common among non-responders to MTX treatment suggesting that Pakistani patients with this genotype are less likely to benefit from MTX