Identification of Variants and Mutational Analyses of Cardiac Myosin-binding Protein C (MYBPC3) Gene of Adult Bangladeshi Patients with Hypertrophic Cardiomyopathy.

Hypertrophic cardiomyopathy (HCM) is the most prevalent genetic hereditary cardiomyopathy characterized by sudden cardiac death. Mutations in the MYBPC3 gene are often the most prevalent genetic abnormality in HCM with a prevalence ranging from 20.0 to 42.0%. The mutation spectrum is available for different countries, but such studies are lacking in the Asian population including Bangladeshi patients. A cross-sectional descriptive study was conducted for mutation analysis of the whole MYBPC3 gene on a cohort of 75 HCM Bengali Bangladeshi probands through Next Generation Sequencing at the Genetic Research Lab of Bangabandhu Sheikh Mujib Medical University from 2016 to 2019. The structural and functional impact of the mutations was further analyzed by in silico process. We analyzed the data and found 103 variants in 102 locations in the MYBPC3 gene. Variants were identified in both the coding region and the non-coding region. We found one possibly novel variant in the MYBPC3 gene. The findings of this research will help to develop a genetic database of HCM which will help in the early diagnosis and proper management of HCM patients in Bangladesh. One pathogenic splice donor variant (47356592 C >T) was found in the intronic region. Among the variants in the coding region, one missense mutation was pathogenic (NP₋000247.2: p.Asp770Asn) which was found in seven patients and another one is of conflicting interpretations of pathogenicity (NP₋000247.2: p.Ser217Gly) which was found in two patients. We have identified one in-frame deletion (NP₋000247.2: p.Ala433del) that is possible a novel variant responsible for the development of HCM.

Estimation of Serum Creatinine and Uric Acid in Bangladeshi Gestational Diabetic Mother Attending in Tertiary Care Hospital.

The hyperuricaemia in Gestational diabetes mellitus has been explained to be a component of the metabolic syndrome which reflects insulin resistance and it has been shown to have a positive correlation with the creatinine levels. Gestational hyperuricaemia was found to be significantly associated with a high rate of maternal and foetal complications along with proteinuria and hypertension. Aimed of this study was to evaluate the serum creatinine and uric acid levels in Bangladeshi women with GDM in their second and third trimester of pregnancy. This descriptive cross sectional study was conducted at Mymensingh Medical College Hospital (MMCH), Mymensingh, Bangladesh from July 2013 to June 2014. Pregnant women, in their second and third trimester, attending the outpatient department of Obstetrics and Gynecology and the Department of Endocrinology, MMCH, Mymensingh, Bangladesh were enrolled by purposive sampling technique. GDM was diagnosed on the basis of oral glucose tolerance test (OGTT) as outlined in UN agency WHO criteria 2013. Out of 172 participants, 86 had GDM (Case) and 86 were normoglycemic (control). The mean age of GDM and control groups was 28.6±3.2 and 27.3±3.1 years respectively. The BMI was 26.4±1.5 kg/m² and 26.3±1.3 kg/m². In this study we found serum creatinine levels in GDM cases were significantly (p<0.001) increased in both trimesters (1.09±0.16mg/dl and 1.07±0.11mg/dl) compared to those without GDM (0.69±0.16mg/dl and 0.64±0.15mg/dl). Serum uric acid levels in GDM and without GDM in 2nd trimester were (4.47±0.42mg/dl and 4.43±0.63mg/dl respectively) had (p>0.05) no significant difference. Serum uric acid levels in GDM cases were significantly (p<0.001) higher in third trimesters (4.48±0.41mg/dl) compared to those without GDM (3.52±0.74mg/dl). There was distinct alteration of serum creatinine and uric acid levels in GDM compared to normal pregnancy.

Estimation of Serum Zinc, Copper and Magnesium Levels in Bangladeshi women with Gestational Diabetes Mellitus Attending in a Tertiary Care Hospital.

Zinc (Zn), Copper (Cu) and Magnesium (Mg) are essential trace elements for normal embryogenesis and fetal growth. Alteration of Zn, Cu and Mg concentrations in blood has been observed in normal pregnancy as well as in gestational diabetes mellitus (GDM). Aim of this study was to evaluate the serum Zn, Cu and Mg levels in Bangladeshi women with GDM in their second and third trimester of pregnancy. This descriptive cross sectional study was conducted at Mymensingh Medical College Hospital (MMCH), Mymensingh, Bangladesh from July 2013 to June 2014. Pregnant women, in their second and third trimester, attending the outpatient department of Obstetrics and Gynecology and the Department of Endocrinology of MMCH were enrolled by purposive sampling technique. GDM was diagnosed on the basis of oral glucose tolerance test (OGTT) as defined in WHO criteria 2013. Out of 172 participants, 86 had GDM (Case) and 86 were normoglycemic (control). The mean age of GDM and control groups was 28.6±3.2 years and 27.3±3.1 years respectively. The BMI was 26.4±1.5kg/m² and 26.3±1.3kg/m². In this study we found serum Zn levels in GDM cases were significantly (p<0.001) low in both trimesters (43.93±5.48µg/dl and 46.86±3.12µg/dl) compared to those without GDM (67.30±7.81µg/dl and 67.58±9.12µg/dl). On the contrary, serum Cu levels in GDM cases were significantly (p<0.001) higher in both trimesters (224±3.8µg/dl and 243.91±6.9µg/dl) compared to those without GDM (220.1±7.6µg/dl and 234.9±4.6µg/dl). There was significant (p<0.001) increase of serum Cu levels in 3rd trimester compared to 2nd trimester in both GDM and non GDM cases. Serum Mg level was significantly low (p<0.001) in 2nd and 3rd trimesters in GDM cases (1.39±0.26mg/dl and 0.93±0.15mg/dl) compared to control group (1.67±0.30mg/dl and 1.67±0.31mg/dl). There was distinct alteration of serum Zn, Cu and Mg levels in GDM compared to normal pregnancy.

Study of Thyroid Disorders among Type 2 Diabetic Patients Attending a Tertiary Care Hospital.

Type 2 diabetes mellitus (T2DM) and thyroid disorders (TD) are the two most common endocrine disorders in clinical practice. Unrecognized thyroid disorders may adversely affect the metabolic control and add more risk to an already predisposing type 2 diabetes mellitus. The objective of the study is to find out the percentage of hypothyroidism and hyperthyroidism in patients with type 2 diabetes mellitus and non diabetic subjects. This cross-sectional study was conducted in the department of Biochemistry and outpatient Department of Endocrinology, BIRDEM General Hospital, Dhaka, Bangladesh from July 2014 to June 2015. Total 200 patients were selected by inclusion and exclusion criteria. Among them, 115 were T2DM subjects and 85 were non diabetic subjects. Type 2 diabetes mellitus and normoglycemia were diagnosed according to WHO criteria from previous medical records. All patients were underwent thyroid function tests; free thyroxine (FT4), free tri-iodothyronine (FT3) and S. TSH (thyroid stimulating hormone). Study sample were divided 4 groups: Group 1: Type 2 diabetic subjects with thyroid disorders; Group 2: Non diabetic subjects with thyroid disorders; Group 3: Type 2 diabetic subjects without thyroid disorders; Group 4: Non diabetic without thyroid disorders. Thyroid disorders were categorized into subclinical hypothyroidism, overt hypothyroidism, subclinical hyperthyroidism and overt hyperthyroidism. Percentage of thyroid disorders in type 2 diabetes mellitus and non diabetic patients were estimated. Comparison of all demographic characteristics was done among the groups. The percentage of thyroid disorders in type 2 diabetic patients was 23.5%. The most frequent thyroid disorders in type 2 diabetic patients were subclinical hypothyroidism (10.4%) and overt hypothyroidism (6.1%). The percentage of thyroid disorders in non diabetic subjects was 12%. Here, subclinical hypothyroidism is 5.9% and overt hypothyroidism is 3.5% were significantly higher in female, overweight and obese subjects. Patients with thyroid disorders (both type 2 diabetic and non-diabetic) had strong association with family history of thyroid disorders. This study showed that the percentage of thyroid disorders in type 2 diabetes mellitus is very high. Specially hypothyroidism (both subclinical and overt) are most frequent. So it is very necessary to provide regular screening for thyroid disorders in type 2 diabetic patients as well as initiating early treatment. Early treatment thyroid disorders in type 2 diabetic patients will help us to avoid complications and reduce morbidity and mortality of diabetes mellitus and thyroid disorders.

Challenges in HbA1c Analysis and Reporting in Patients with Variant Hemoglobins.

Hemoglobin A1c (HbA(1)c) is a well-established indicator of mean glycemia. The presence of genetic variants of hemoglobin can profoundly affect the accuracy of HbA(1)c measurements. Variants of hemoglobin especially Hemoglobin E (HbE) is prevalent in South East Asia including Bangladesh. The objective of our study is to compare the HbA(1)c values measured on high performance liquid chromatography (HPLC) and Turbidimetric Inhibition Immunoassay (TINIA) in diabetic patients with variant hemoglobins including HbE. A total of 7595 diabetic patients receiving treatment at BIRDEM General Hospital were analyzed for HbA(1)c results within a period of two months from December 2013 to January 2014. Seventy two cases out of 7595 (0.95%) had either undetectable or below normal HbA(1)c levels (males-33 and females-39; ratio = 0.82:1) by HPLC method. In 34(0.45%) cases, HbA(1)c value was undetectable by HPLC method but was in the reportable range by TINIA method. In the other 38 (0.55%) cases, HbA(1)c levels were below the reportable range (<4%) by HPLC method but were in the normal or higher range by TINIA method. TINIA method did not agree with HPLC method on Bland Altman plot in the 38 cases with below normal HbA(1)c levels, [Mean bias -5.2(-9.3 to 1.0), 95% CI] but agreed very well [mean bias -0.21 (-0.84 to 0.42), y=1.1037+0.776X; r(2)=0.30, p<0.01] in controls. In control group mean MCV was 83.80±7.48 and in study group was 73.65±10.44. Alkaline electrophoresis confirmed the variant hemoglobin to be HbE. The fasting blood sugar levels of all the 72 cases correlated strongly with TINIA method (r(2) =0.75, p<0.0001) but not with HPLC (r = 0.24, p=0.13). In our regions where populations have a high prevalence of Hb variant, proper knowledge of hemoglobin variants which affect the measurements HbA(1)c level is essential. MCV of 80fl or below may serve as a rough guide to select samples that require analysis by TINIA method. Moreover, HPLC may be a convenient and inexpensive tool for screening of hemoglobinopathies especially among diabetic population in Bangladesh. It may therefore be helpful in improving management of complications related to both anaemia and iron overload.

The Aspect of NK2 Transcription Factor Related Locus-5 (NKX2.5) Gene Mutations in Bangladeshi Atrial Septal Defect (ASD) patients and 2D Relationship with their Age.

Atrial septal defect (ASD) is a developmental defect of the heart which arises from the congenital abnormality of interatrial septum that perturbs the normal blood flow. Development of the heart is a complex biological process regulated by numerous genetic and environmental factors. During this process DNA binding proteins Myocardin, NKX2.5 (NK2 Transcription Factor Related Locus-5) and GATA4 (GATA Binding Protein-4) function by binding to SRF (Serum Response Factor) which is also a key regulator of myogenic terminal differentiation and frequently results in mitogenesis. Several studies suggest that mutations in the homeodomain containing transcription factor, NKX2.5, is implicated with atrial septal defect. This cross sectional descriptive study was done to investigate the frequency of NKX2.5 gene mutations among the patient with ASD who were undergoing surgical repair at the National Institute of Cardiovascular Diseases (NICVD) and National Heart Foundation and Research Institute (NHF&RI), Dhaka from July 2010 to June 2011. Patients presented with ASD at any age of both sexes were selected as study population. We found six distinct polymorphic sites among Bangladeshi population. Among six polymorphic sites, two were located at position 487 and 495. These were present in around 80% of the affected individuals. However they were not present in control population. Our study also revealed that mutations present in the downstream sites or towards the end of the genes are restricted to older people, whereas mutations present towards the 5' site is common to population of all ages. This interesting relationship has encouraged us to raise two new hypotheses.

Value of red cell distribution width (RDW) and RBC indices in the detection of iron deficiency anemia.

Iron deficiency anemia is the common problem during pregnancy. Detection of iron deficiency early during pregnancy can reduce maternal and child mortality and morbidity. Red cell distribution width is a new routine parameter in fully automated hematology analyzer that can give the idea of early iron deficiency before other test. It gives the idea of red cell size variation which is the earliest morphologic changes in iron deficiency anemia. In prelatent and latent stage of iron deficiency MCV are normal. Whereas in latent stage Red Cell Distribution Width (RDW) would be expected to increase because of a microcytic population of cells appears in the blood. This study evaluates the role of red cell distribution width and RBC indices in determining iron deficiency early and provide reliable and useful technique. In this study 190 pregnant women were included. RDW, MCV, MCH, MCHC and iron profile were done. RDW compared with MCV, MCH and MCHC in various stages of iron deficiency. In latent stage of iron deficiency higher RDW was found significant than MCV, MCH, MCHC (p<0.05). In this study RDW had sensitivity 82.3% and specificity 97.4%. Whereas MCV, MCH and MCHC had 29.2%, 68.1% and 15% sensitivity but specificity was 98.7%, 83.1% and 96.1% in the detection of iron deficiency. Iron deficiency anaemia without other complicating disease could be screened out early by increased RDW when RBC indices were normal.

Role of red cell distribution width (RDW) in the detection of iron deficiency anaemia in pregnancy within the first 20 weeks of gestation.

Iron deficiency anemia is common problem during pregnancy. Red cell size variation (anisocytosis) is the earliest morphologic changes in iron deficiency anemia. Red cell distribution width is a quantitative measure of red cell size variation and it can give the idea of early iron deficiency before other test to become positive. 190 pregnant women were included in this study. Red cell distribution width was compared between iron deficient & non-iron deficient pregnant women. Red cell distribution width also compared with Hb level, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration and peripheral blood film in prelatent iron deficiency, latent iron deficiency, mild and moderate iron deficiency anemia. Red cell distribution width had sensitivity 82.3% and specificity 97.4%. Whereas Hb level, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration and peripheral blood film all had 56.6%, 29.2%, 68.1%, 15% and 38.9% sensitivity but specificity was 90.9%, 98.7%, 83.1%, 96.1% and 98.7% in the detection of iron deficiency. Red cell distribution width appears to be a reliable and useful parameter for detection of iron deficiency during pregnancy.

Drug utilization of oral hypoglycemic agents in a university teaching hospital in India.

BACKGROUND: India has witnessed a rapidly exploding epidemic of diabetes in recent years and currently leads the world with the largest number of diabetic subjects in a single country. World Health Organization estimates that in 2000, 31.7 million individuals were affected by diabetes in India and these numbers will rise to 79.4 millions by the year 2030. In view of the above situation, drug utilization review of antidiabetic medicines in Indian healthcare settings has a valid significance to promote rational drug use in diabetics. OBJECTIVE: The present study is aimed to determine the drug utilization patterns in type 2 diabetic patients on oral hypoglycemic agents in the Medicine Outpatient Department (OPD) and Inpatient Department (IPD) of Majeedia Hospital, a teaching hospital of Hamdard University, New Delhi. METHODS: Patients with established type 2 diabetes (n = 218) visiting the OPD and IPD were interviewed using a structured questionnaire during the period January-May 2006. RESULTS: A majority of the type 2 diabetic patients in this setting were treated with multiple antidiabetic drug therapy. The most commonly prescribed antidiabetic drug class was biguanides (metformin) followed by sulphonylureas (glimepiride), thiazolidinediones (pioglitazone), insulin and alpha-glucosidase inhibitors (miglitol). As monotherapy insulin was the most common choice followed by metformin. The most prevalent multiple therapy was a three-drug combination of glimepiride + metformin + pioglitazone. More than half of the type 2 diabetic patients showed poor adherence (compliance) to the prescribed therapy. CONCLUSION: This study strongly highlights the need for patient education or counselling on use of antidiabetic and concomitant drugs, monitoring of blood glucose and glycosylated haemoglobin (HbA1c) levels, diet control, and correction of diabetic complications. Metabolic control was poor and HbA1c monitoring was underutilized. Clinical monitoring of patients' adherence to prescribed treatments is recommended and measures should be taken to improve it.

Effects of inorganic phosphorus compounds on the hydrolysis of phosphatidylcholine liposomes by phospholipid-deacylating enzymes.

Structural requirements of inorganic phosphorus compounds as specific activators or inhibitors for phospholipase A2 and phospholipase B were investigated using orthophosphate, pyrophosphate and polyphosphate. It was observed that orthophosphate and pyrophosphate stimulated the activities of phospholipase A2 from bee venom, snake (Naja naja) venom and pig pancreas, and also phospholipase B from the yeast Torulaspora delbrueckii. However, polyphosphate was found to act as an inhibitor for phospholipase A2 in the above species and also for phospholipase B from T. delbrueckii. Orthophosphate and pyrophosphate induced gradual aggregation of liposome, but polyphosphate prolonged the lifetime of the liposome, suggesting that orthophosphate and pyrophosphate destabilize the bilayer structure of phosphatidylcholine and polyphosphate stabilizes it.

Cloning and sequencing of phospholipase B gene from the yeast Torulaspora delbrueckii.

The extracellular phospholipase B gene from baker's yeast Torulaspora delbrueckii was cloned and sequenced. Analysis of DNA sequence data revealed an open reading frame (ORF) encoding a 649-amino acid protein, that included amino acid sequences obtained from the purified enzyme. Comparison of these sequence data with the N-terminal amino acid sequence of the enzyme indicated that this secreted protein is synthesized as a large precursor with a 21-amino acid N-terminal extension to the mature enzyme of 628 amino acids. A homology search was carried out between phospholipase B from T. delbrueckii and Penicillium notatum. The deduced amino acid sequence of the cloned phospholipase B was homologous (about 50% identity) to phospholipase B from P. notatum, and contained six conserved regions. The transcriptional level of mRNA of the phospholipase B gene was higher in the cells from early exponential and stationary phases.

Calcium ion dependent interaction of inorganic phosphorus compounds with phosphatidylcholine.

The translocation of orthophosphate, pyrophosphate, and polyphosphate into an organic phase was examined by means of the two-phase partition method. When the organic phase contained phosphatidylcholine (PtdCho), the phosphorus compounds translocated from the water-phase into the organic phase, depending on the concentrations of PtdCho and Ca2+, and the pH of the solution. Calcium ions were essential for the translocation. The concentration of Ca2+ necessary for the translocation appeared to depend on the molar concentration of the phosphorus compounds. Orthophosphate was translocated above pH 6.5, while pyrophosphate was translocated above pH 3.5. In contrast, polyphosphate was translocated in the whole pH range examined (pH 2.5-9.0), although approx. 9% of the total polyphosphate remained in the water-phase even at alkaline pH. These results suggest that phosphorus compounds can interact with PtdCho when the phosphorus compounds have formed complexes with Ca2+.