Expression of neutrophil elastase and myeloperoxidase mRNA in patients with newly diagnosed type 2 diabetes mellitus.

BACKGROUND AND AIMS: Neutrophil elastase and myeloperoxidase enzymes protect us from infection by killing pathogens. However, exaggerated activities of these enzymes can induce tissue damage, inflammation and oxidative stress. The present study was aimed to explore the expressions of neutrophil elastase and myeloperoxidase mRNA in the peripheral blood leukocytes (PBL) in patients with newly diagnosed type 2 diabetes mellitus. METHODS: In this cross-sectional study, 104 participants including 65 normoglycemic control subjects and 39 newly diagnosed type 2 diabetes patients were recruited. Glycemic and metabolic markers were evaluated from fasting blood samples. The mRNA levels of neutrophil elastase and myeloperoxidase genes in the PBL were quantified by real-time quantitative PCR. RESULTS: Compared to control subjects, diabetes patients showed a significant down regulation of both neutrophil elastase (p = 0.039) and myeloperoxidase (p = 0.023) mRNA expressions in the PBL. The neutrophil elastase and myeloperoxidase mRNA levels showed a negative trend with fasting glucose levels but did not show any significant correlations with HbA1c, insulin level, insulin resistance or sensitivity status. CONCLUSIONS: It was concluded that type 2 diabetes mellitus is associated with a decrease in neutrophil elastase and myeloperoxidase gene expression in the PBL.

Neutrophil elastase and myeloperoxidase mRNA expression in overweight and obese subjects.

Neutrophil elastase and myeloperoxidase enzymes have been implicated in high-fat diet-induced obesity, insulin resistance (IR) and atherosclerosis in animal models. The aim of the present study was to explore neutrophil elastase and myeloperoxidase mRNA expressions in the peripheral blood leukocytes (PBL) in overweight and obese subjects, and to correlate those mRNA expressions with BMI, IR and cardiovascular biomarkers. In this cross-sectional study, 74 apparently healthy subjects including 22 lean, 27 overweight and 25 obese subjects were recruited. Cardiovascular and metabolic biomarkers were evaluated from fasting blood samples. The mRNA levels of neutrophil elastase and myeloperoxidase genes in the PBL were quantified by real-time PCR. Compared to lean group, the overweight and obese groups showed significant upregulation of both neutrophil elastase (p < 0.001) and myeloperoxidase (p < 0.03) mRNA expressions in the PBL. But no difference was found between overweight and obese groups. The neutrophil elastase and myeloperoxidase mRNA levels showed significant positive correlation with BMI, serum triglyceride, atherogenic index of plasma and 10-year risk of developing cardiovascular disease. But no correlation was found with glucose, insulin or IR. It was concluded that the neutrophil elastase and myeloperoxidase genes are up-regulated in both overweight and obese subjects and are associated with BMI and markers of cardiovascular disease.

Effect of oxidative stress in male infertility.

Infertility is a medical and social problem all over the world. Infertility results from abnormality of the male partners in almost 50% of cases. Oxidative stress is involved with many chronic pathological conditions and the current study was designed to evaluate any association that may exist between male infertility and oxidative stress. Infertile male patients (having female partners with normal fertility parameters; n=31) and age- matched healthy male fertile control subjects (n=30) were randomly selected from the Infertility Unit of the Gynaecology and Obstetrics Department of Bangabandhu Sheikh Mujib Medical University. As a marker of oxidative stress-induced lipid peroxidation, erythrocyte and seminal plasma malondialdehyde (MDA) levels were measured in fertile and otherwise healthy infertile male subjects. In addition, antioxidant capacity was evaluated by measuring erythrocyte and seminal plasma glutathione (reduced form, GSH) concentrations in fertile and infertile male subjects. The MDA and GSH levels were measured by thiobarbituric acid method and Elmans method respectively. The median (range) MDA level of erythrocyte was found significantly higher in infertile male subjects compared with healthy fertile male subjects 24.23(3.71-42.21) vs. 12.84(1.30-24.00)nmol/gm Hb p<0.001. However, the erythrocyte GSH level did not differ between the two groups 12.62(0.67-29.82) versus 13.93 (2.10-21.08)mg/gm Hb. In case of seminal plasma, the median (range) MDA level was found significantly higher in infertile group 3.17 (1.20-6.21) versus 1.88(0.50-5.37)nmol/ml, p<0.001. In addition, the seminal plasma GSH level was found markedly suppressed in infertile group compared with fertile group 1.64(0.23-7.50) versus 4.26(2.32-7.50)mg/dl, p<0.001. Taken together, infertile male patients show an elevation of oxidative stress markers both in the erythrocytes and in the seminal plasma. Indicating that male infertility might be associated with increased oxidative stress.

Role of cardiac troponin I in staging and prognosis of congestive heart failure subjects.

Congestive heart failure is the one of the major cardiovascular disorder that is increasing in incidence and cause of death globally. Mortality rate has increased 40%-50% in advanced cardiac failure and 15%-25% in mild to moderate cardiac failure within one year of diagnosis. There is no established biochemical marker for the diagnosis, prognosis and staging of heart failure. Cardiac Troponin I may be a novel useful tool in identifying patients with Heart failure who are at increased risk for progressive ventricular dysfunction and death. Thirty six congestive heart failure cases and thirty six healthy controls were included in this study and serum cardiac troponin I and Ejection fraction were measured. All the study subjects were grouped according to the NYHA class they belong. Cardiac troponin I was significantly higher in CHF cases than the controls. Troponin I also significantly differed among groups. EF of cases was significantly lower than the controls and also differed among groups. A significant negative correlation between cardiac troponin I and progressive decline of ejection fraction was evident in this study. Cardiac troponin I increased progressively with progression of heart failure. Thus, Cardiac troponin I could be used to stratify patients undergoing heart failure in to high and low risk groups for future cardiac events. Cardiac troponin I could also be used as a very important marker for the prognosis of the patients with congestive heart failure.

Spot morning urine protein creatinine ratio and 24 hour urinary total protein excretion rate.

Proteinuria helps to establish the diagnosis of most renal diseases and also to predict the outcome of such diseases. Proteinuria is biochemically represented by measuring the protein concentration in timed collection of 24 hour urine. But, 24-hour timed urine collection is time consuming, cumbersome and often unreliable due to collection errors and also results in undue delay on diagnostic process. An alternate approach avoiding arduous and inaccurate timed urine collection can be the measurement of protein creatinine ratio in spot morning urine. This study was aimed to evaluate whether the spot morning urine protein creatinine ratio can be a reliable alternative to 24-hour urinary total protein (UTP) estimation. The study was carried out in the department of Biochemistry, Bangabandhu Sheikh Mujib Medical University, Dhaka on 50 (fifty) non-diabetic Chronic Renal Disease (CRD) patients with an age ranging from 18 -70 years. The study subjects were grouped into mild, moderate and severe CRD on the basis of GFR. Urinary protein and creatinine concentrations were measured in spot morning urine samples and their ratios were calculated. Urinary protein measured in 24-hour timed collected urine samples gave the 24-hour UTP excretion rate. In our study, spot morning urine protein creatinine ratio significantly correlated with 24-hour UTP excretion rate in all CRD patients. Severe CRD patients gave significant positive correlation (p<0.05), whereas mild and moderate CRD patients gave very highly significant positive correlation (p<0.001). Therefore, it may be suggested that protein creatinine ratio in spot morning urine can be accepted as a reliable and alternative to 24-hour UTP excretion rate in non-diabetic chronic renal disease patients. This simple and inexpensive procedure will thus simplify the way of establishing the severity of renal disease along with its prognosis.

Lipoprotein -- an independent risk factor for CVD.

A case control study done to evaluate the Lipoprotein(a) [LP(a)] as a risk factor for CVD (cerebrovascular disease). 150 non-smokers, non-alcoholic subjects free from DM, renal disease, thyroid disease and liver disease were included in the study. Among them 120 were CVD cases and 30 were age and sex matched healthy control. Subjects were grouped as group-I (30, healthy control), Group-II (60, Hemorrhagic CVD) and group-III (60, Ischemic CVD). Fasting (12 hr) blood samples were collected from all subjects and in CVD cases samples were collected after 24 hr of attack. Lipid profile and LP(a) conc. were measured in all samples. Mean serum LP(a) conc. in Group-I, Group-II and Group-III were found to be 17.6 7.4 mg/dl, 31.9 15.6 mg/dl and 44.8 24.0 mg/dl respectively. Both the groups of CVD cases showed significantly higher level of serum LP(a) conc. compared to healthy control. CVD cases did not differ statistically in respect of their lipid profile when compared with control. Moreover the serum LP (a) conc. of CVD cases found to show no correlation with their lipid profile, suggesting the serum LP(a) conc. a possible independent risk factor for CVD.

Serum homocysteine concentration in patients with acute MI and chronic IHD.

This was an observational study carried out in the Department of Biochemistry, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka; with the active co-operation of Coronary Care Unit (CCU) of BSMMU & National Institute of Cardiovascular Disease (NICVD), Dhaka. This study was carried out from March 2002 to January 2003. Total seventy (70) subjects were studied. Out of them 20 were of Acute MI, 20 were Chronic ischemic heart disease (CHD) and 30 were age and sex matched healthy controls. Hospitalized diagnosed patients were selected by taking history, clinical examination and several investigations like ECG. Echocardiogram, Angiogram and several enzymes assay. Several studies in many countries showed that serum homocysteine (Hcy) was elevated in IHD Patient. Cardiovascular disease is alarmingly increasing in Bangladesh. So our aim and objective of the study was to find out the association of serum Hcy with Acute MI and chronic ischemic heart disease (CHD) patients in our population. Mean Hcy level of Acute MI were 21.16 +/- 4.56 (micromol/l), 27.55 +/- 10.40 (micromol/l) and that of control was 13.03 +/- 10.51(micromol/l). Serum Hcy was significantly higher in both cases than control. But insignificant difference was found between AMI vs CHD (P> 0.05). Quantitative measurement of serum Hcy was measured by fluorescence polarization Immunoassay (FPIA) in IMX analyzer (Abbott-USA).

Lipoprotein-a: a laboratory tool for clinical categorization of CVD.

A case-control study was done to evaluate the association of Lipoprotein(a)[LP(a)] with CVD (Cerebrovascular disease) and also to assess the implication of serum LP(a) concentration as a differentiating marker between ICVD (Ischemic CVD) & HCVD (Hemorrhagic CVD). 150 non-smokers, non-alcoholic subjects free from DM, renal disease, thyroid disease and liver disease were studied. Among them 120 were CVD cases and 30 were age & sex matched healthy control. Fasting (12 hr.) blood samples were collected from all subjects and in CVD cases samples were collected after 24 hr. of attack. Serum LP(a) concentration were measured in all samples. Mean serum LP(a) concentration in control, HCVD & ICVD were found to be 17.6 +/- 7.4 mg/dl, 31.9 +/- 15.6 mg/dl and 44.8 +/- 24.0 mg/dl respectively. Both HCVD & ICVD cases showed significantly higher level of serum LP(a) concentration compared to control. Moreover ICVD cases showed significantly higher level of serum LP(a) concentration compared to HCVD cases. The exquisite athero-thrombo-embolic potential of LP(a) explain its involvement with CVD but more with ICVD in comparison to HCVD; This finding apparently suggest the prospect of serum LP(a) concentration to be used as a promising laboratory maker to differentiate clinically the ICVD from HCVD following determination of cut-off value between ICVD & HCVD by broad based comprehensive study.

Studies on serum triglyceride level in patients with myocardial infarction.

Myocardial Infarction (MI) is the most common form of heart disease and the single most important cause of premature death in the developed and developing world. Unfortunately the incidence of the condition is increasing rapidly in many developing countries like Bangladesh. Effort should therefore be taken to minimize the risk factors of MI. Large scale randomized clinical trials have shown that lowering high triglyceride concentration mainly by drugs reduces the risk of cardiac events like MI. So the present work has been designed to see the serum triglyceride levels in normal healthy subjects, to compare serum triglyceride levels in patients with MI and those of healthy subjects and to evaluate the association of serum triglyceride in Bangladeshi MI patients. The present study was carried out in the Department of Biochemistry, BSMMU in collaboration with Department of Cardiology, BSMMU and NICVD, Dhaka during the period of July 2001 to December 2002. A total of 50 subjects were selected, Group A (30 subjects of control ) and Group B (20 subjects of test ). The mean level of serum triglyceride in control subjects were 117.07 +/- 32.41 mg/d1 and in test subjects were 176.87 +/- 37.15 mg/d1. So the present study showed that serum triglyceride level is significantly higher in patients with MI. From the present study, it is difficult to draw any definite conclusion but suggested that high serum triglyceride concentration is a cause of the incidence of MI.

Glycemic status in hyperthyroid subjects.

This was an observational case-control study carried out in the Department of Biochemistry, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka jointly with the 'Thyroid and Endocrine Clinic' of the same institution during the period of January 2002 to December 2002. Sixty-five (65) newly diagnosed hyperthyroid patients between 20-60 years of age were studied, where forty-five (45) were Graves' disease and twenty (20) were TMNG (Toxic multinodular goiter) patients. Thyrotoxicosis was diagnosed by history, clinical examination and biochemical investigations- FT4, TSH, and Radioactive iodine uptake (RAIU) test. Thirty (30) age and sex matched healthy subjects were taken as control. The mean age was 33.02+/-9.24 years in Graves' disease and 37.55+/-9.49 years in TMNG. Female predominance observed in both the diseases. Glucose intolerance was found in 72.3% of thyrotoxic patients, which is much higher than European population. Our study showed Diabetes mellitus (DM) in 11% of Graves' disease patients. The incidence of DM in Graves' disease was slightly higher in our population. Incidence of DM in TMNG in our study was much lower (5%) than that of Graves' disease (11%) but the incidence of IGT (Impaired glucose tolerance) in TMNG was more (85%) in relation to Graves' disease (54%). Percentage of RAIU was more marked in Graves' disease than TMNG. There is a significant positive correlation (p<0.05) between plasma glucose and FT4 in Graves' disease. Glucose intolerance is frequently found in Thyrotoxic patients.

Characterisation of Jack fruit lectin.

Jack fruit (Artocarpus Heterophyllus) seed extract contains a lectin termed Jack fruit lectin (JFL) which possesses diversed biological properties. A detailed analysis of its properties has been lacking. The present investigation was initiated to study the detail properties of JFL. After extraction and purification on affigel galactosamine-agarose column, JFL was subjected to ND-PAGE. Several different charged species from ND-PAGE upon SDS-PAGE gave rise to two dissimilar trimeric subunit at 12.5 and 15.0 KDa and retain biological activity. It was possible to elute the subunit bands separately from polyacrylamide gel to investigate their biological activity. Each subunit was found to be retained the lectin activity. Agglutinating activity of smaller subunit was found to be more, may be due to the greater amount of the subunit. This also suggests that each unit of trimeric JFL have similar lectin activity.

Isolation modifies the behavioural response in rats.

Twenty-four male wister rats were reared from weaning either alone (isolation reared) or in groups of five (socially reared) for 6 weeks. Thereafter, the present study examined and compared the behaviour of isolation and socially reared rats in the test of social interaction, in the elevated plus-maze test of anxiety, in the open field behavior of exploration and locomotion activity, and the depressive behaviour in forced swim test. Under high light in unfamiliar conditions isolation reared rats spent significantly more time in aggressive interactions (P < 0.05) and high levels of aggressive behaviour compared to the socially reared rats (P < 0.05). Isolation reared rats had spent less time in the open arms than the socially reared rats in the maze test (P < 0.05). Isolation reared rats were more exploratory than the socially reared rats in the open field test and the defaecation scores were less compared to the socially reared controls (P < 0.05). Isolated rats spent less time immobile on the rats' forced swim test behaviour but the difference was not statistically significant. These findings suggest that isolation reared rats are nervous, aggressive and hyperactive animals in a novel field. It appears that isolation rearing in the early stages of life has modified a variety of behaviour in the adult rats, and the investigation of the pattern of behavioural changes in isolation reared rats may help to explore the environmental influences on the development of human psychopathology.