High-resolution melt curve analysis: An approach for variant detection in the TPO gene of congenital hypothyroid patients in Bangladesh.

TPO (Thyroid Peroxidase) is known to be one of the major genes involved in congenital hypothyroid patients with thyroid dyshormonogenesis. The present study aims to validate high-resolution melting (HRM) curve analysis as a substitute method for Sanger sequencing, focusing on the frequently observed non-synonymous mutations c.1117G>T, c.1193G>C, and c.2173A>C in the TPO gene in patients from Bangladesh. We enrolled 36 confirmed cases of congenital hypothyroid patients with dyshormonogenesis to establish the HRM method. Blood specimens were collected, and DNA was extracted followed by PCR and Sanger sequencing. Among the 36 specimens, 20 were pre-sequenced, and variants were characterized through Sanger sequencing. Following pre-sequencing, the 20 pre-sequenced specimens underwent real-time PCR-HRM curve analysis to determine the proper HRM condition for separating the three variations from the wild-type state into heterozygous and homozygous states. Furthermore, 16 unknown specimens were subjected to HRM analysis to validate the method. This method demonstrated a sensitivity and specificity of 100 percent in accurately discerning wild-type alleles from both homozygous and heterozygous states of c.1117G>T (23/36; 63.8%), c.1193G>C (30/36; 83.3%), and c.2173A>C (23/36; 63.8%) variants frequently encountered among 36 Bangladeshi patients. The HRM data was found to be similar to the sequencing result, thus confirming the validity of the HRM approach for TPO gene variant detection. In conclusion, HRM-based molecular technique targeting variants c.1117G>T, c.1193G>C, and c.2173A>C could be used as a high throughput, rapid, reliable, and cost-effective screening approach for the detection of all common mutations in TPO gene in Bangladeshi patients with dyshormonogenesis.

Hypothalamic-Pituitary-Adrenal Axis Activity in SARS-CoV-2 Infected Noncritically Ill Hospitalized Patients.

Objectives: This study determined the baseline hypothalamic-pituitary-adrenal axis hormonal levels and their associated factors in noncritically ill hospitalized patients with coronavirus disease 2019 (COVID-19). Methodology: This cross-sectional study was carried out in 91 noncritical RT-PCR-confirmed COVID-19 patients (aged 18 to 65 years) recruited consecutively from the COVID unit of two tertiary care hospitals over a period of six months. After the screening, relevant history and physical examinations were done, and blood was drawn between 07:00 am to 09:00 am in a fasting state to measure serum cortisol and plasma adrenocorticotropic hormone (ACTH) by chemiluminescent microparticle immunoassay. Results: Of 91 patients, 54, 26, and 11 had mild, moderate, and severe COVID-19, respectively. Median values of serum cortisol (p = 0.057) and plasma ACTH (p = 0.910) were statistically similar among the severity groups. Considering a cortisol cut-off of 276 nmol/L (<10 µg/dL), the highest percent of adrenal insufficiency was present in severe (27.3%), followed by mild (25.9%) and least in the moderate (3.8%) COVID-19 cases. Using the cortisol/ACTH ratio >15, only 6.6% had enough reserve. Conclusions: The adrenocortical response was compromised in a significant percentage of noncritically ill hospitalized patients with COVID-19, with the highest percentage of adrenal insufficiency present in severely infected cases. The HPA axis parameters of serum cortisol, plasma ACTH and cortisol/ACTH were similar across the severity of noncritical patients with COVID-19.

Sex Reversal Syndrome (SRS): A Case of SRY-Positive 46,XX Testicular Disorder.

We report a case of an SRY-positive 46,XX Indian male who presented with small testis and phallus, poor beard and mustache development and gynecomastia at the age of 24 years. He was biochemically found to have hypergonadotropic hypogonadism. He had 46,XX karyotype and Quantitative Fluorescence-PCR (QF-PCR) identified the SRY gene on the X chromosome. SRY-positive 46 XX male SRS cases usually present as phenotypically male since birth but develop features of hypogonadism, poor testicular development, and infertility after puberty. Infertility, hypogonadism, external genital development, and psychological distress are the major concerns during the management of the patients. Testosterone therapy for hypogonadism, artificial reproductive technologies for fertility, surgical repair of hypospadias/ cryptorchidism/under-virilized genitalia and psychological and genetic counseling are helpful for proper management of the patients.

Lipid Accumulation Product Better Predicts Metabolic Status in Lean Polycystic Ovary Syndrome than that by Visceral Adiposity Index.

Background: Both visceral adiposity index (VAI) and lipid accumulation product (LAP) can be used to assess insulin resistance (IR) and metabolic syndrome (MetS) which are required for management of even lean polycystic ovary syndrome (PCOS) (body mass index [BMI] <23 kg/m2). Aim: This study was aimed to see the magnitude of associations of VAI and LAP with cardiometabolic risk factors including IR and MetS in lean PCOS. Study Setting and Design: This cross-sectional study was done amongst 62 newly detected lean PCOS patients and 58 age- and BMI-matched healthy controls. Materials and Methods: PCOS was diagnosed according to the Revised 2003 Rotterdam Consensus criteria. Along with relevant clinical data, fasting blood was taken to measure glucose, insulin and lipid profile by glucose oxidase, chemiluminescent microparticle immunoassay and by glycerol phosphate dehydrogenase-peroxidase method, respectively. IR was calculated by homeostasis model of IR (HOMA-IR). VAI and LAP were calculated from BMI, waist circumference, triglyceride and high-density lipoprotein cholesterol by using sex-specific formulae. Statistical Analysis Used: Linear and binary regression analyses and receiver operating characteristics curve (ROC) analyses were done as appropriate. Results: Only LAP had predictive associations with age, systolic and diastolic blood pressure and total and low-density lipoprotein cholesterol. Both VAI and LAP had predictive associations with history of subfertility and MetS. LAP had moderate discriminating index for IR with cut-off of HOMA-IR of 2.3. Both VAI and LAP had excellent discriminating index for MetS in lean PCOS patients. Conclusions: LAP had more associations with cardiometabolic risks than VAI and was a moderate discriminator of IR in lean PCOS.

Serum Leptin Correlates with Obesity But Does Not Differ Between Gestational Diabetes and Normal Glucose Tolerance during 24-28 Weeks of Gestation.

Leptin is an adipocytokine secreted by adipocytes which positively correlates with obesity. It is considered as a potential mediator for precipitating Gestational diabetes mellitus (GDM) which is more evident during 24-28 weeks of gestation. This study was conducted to see serum leptin level during 24-28 weeks of gestation in GDM at the Department of Endocrinology, BSMMU, Bangladesh from March 2019 to August 2020. Pregnant women (N=108) were challenged with 75gm oral glucose (OGTT) at 24-28 weeks of gestation and divided into GDM [n=45, age: 27.80±3.98 years, mean±SD; BMI: 27.88 (24.46-30.43) kg/m², median Interquartile range (IQR)] and normal glucose tolerance [NGT; n=62, age: 26.19±5.30 years, mean±SD; BMI: 25.80 (23.65-28.42) kg/m², median (IQR)] on basis of WHO-2013 diagnostic criteria. Fasting serum leptin and glucose were measured by ELISA and glucose oxidase method respectively. No statistically significant difference was found between GDM and NGT for leptin [26.05(16.92-50.55) vs. 23.50(14.95-38.30) median (IQR), p=0.360]. It was also not different statistically between GDM and NGT either for age groups (p=NS for all) or for Asian categories of BMI subgroups (p=NS for all). However, it was higher in subjects with BMI ≥23kg/m² than that with BMI ≤23kg/m² for both GDM [16.65 (6.39, 35.75) vs. 28.35 (19.60, 51.10) median (IQR), p=0.114] and NGT [14.65(9.19, 19.60) vs. 26.00 (17.30, 43.40) median (IQR), p=0.002]. It was also statistically similar in the GDM subgroups divided by Asian BMI cut-off (p=NS). BMI correlated with leptin in NGT (r=0.495, p<0.001) but not in GDM (r=0.177, p=0.251) and regression analysis revealed BMI (kg/m²) as predictor for high leptin (p=0.008). ROC curve analysis for leptin showed AUC for GDM was 0.553 (p=0.360) suggesting it as a poor predictor. It is concluded that fasting leptin in 24-28 weeks of gestation better relates with BMI but does not differ between GDM and NGT anddoes not seem to be a good predictor for GDM. Further study is required to make a comment on its prediction over GDM.

Insulin secretory defect may be the major determinant of GDM in lean mothers.

AIMS: To compare the insulin sensitivity and secretion indices of pregnant Bangladeshi women with GDM and normal glucose tolerance (NGT). METHODS: This cross sectional study was performed with 40 GDM and equal number of NGT pregnant women diagnosed on basis of WHO criterion-2013 during 24-40 weeks of gestation. Glucose was measured by glucose oxidase method and fasting insulin by chemiluminescent immunoassay. Equations of homeostatic model assessment (HOMA) were used to calculate indices of insulin resistance (HOMA-IR), ß-cell function (HOMA-B) and insulin sensitivity (HOMA-%S). RESULTS: The GDM group had significantly higher insulin resistance as indicated by higher fasting insulin value [GDM vs. NGT; 10.23 (7.94-14.50) vs. 7.07 (5.28-11.07) µIU/ml] and HOMA-IR [GDM vs. NGT; 2.47 (1.75-3.43) vs. 1.50 (0.99-2.22)] and poor ß-cell secretion [GDM vs. NGT; HOMA-B: 113.37 (90.30-191.35) vs. 150.98 (109.85-271.72), median (IQR); p < 0.001 for all]. HOMA-B was significantly lower in GDM than NGT with BMI < 23 kg/m2 [GDM vs. NGT; 63.37 (49.19-83.83) vs. 134.89 (93.50-193.17) ng/ml; p = 0.010] despite having statistically comparable difference in IR. BMI was found to be a significant predictor of HOMA-IR in GDM. CONCLUSIONS: Though impairments of both insulin secretion and sensitivity are hallmarks in the pathogenesis of GDM, ß-cell dysfunction contributes more to development of GDM in those with relatively lower BMI in our population.

Antithyroid Antibody Status in Non-Pregnant Adult Bangladeshi Patients with Subclinical Hypothyroidism.

Sub clinical hypothyroidism (SCH) is common in clinical practice. Autoimmunity is thought to be the most important cause of SCH. In this cross-sectional study, we investigated 120 SCH patients and 100 healthy controls attending the Endocrinology Outpatient Department of Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh from June 2014 to April 2015 for anti-thyroid antibodies (anti-TPO and anti-Tg). Measurement of serum TSH, FT4, anti-TPO, and anti-Tg antibodies were done by using the chemiluminescent sequential immunometric assay. SCH patients had a higher mean age; the frequencies of female subjects, those having family history of thyroid disease or other autoimmune diseases, and goiter were higher in SCH group than in the control group. Forty-five percent (45%) of SCH patients were positive for anti-thyroid antibodies (23.3% for both anti-TPO and anti-Tg, 16.7% for only anti-TPO, and 5% positive for only anti-Tg) in comparison to only 10% anti-thyroid antibody positive controls (none for both antibodies, 8% for only anti-TPO, and 2% positive for only anti-Tg). The SCH subjects in the lower age group, females and with a TSH >10µIU/mL had the higher frequency of thyroid autoimmunity. Female gender, high socioeconomic condition, the presence of other autoimmune diseases, the presence of goiter and TSH >10µIU/mL were associated with higher odds of anti-thyroid antibody positivity in the SCH group, though none were statistically significant. The frequency of anti-thyroid antibody was higher in SCH and was more prevalent among the females, younger patients and those having a goiter, other autoimmune diseases, and TSH >10µIU/mL.

Postoperative expression of Cushing disease in a young male: metamorphosis of silent corticotroph adenoma?

SUMMARY: Silent corticotroph adenoma (SCA) is an unusual type of nonfunctioning pituitary adenoma (NFA) that is silent both clinically and biochemically and can only be recognized by positive immunostaining for ACTH. Under rare circumstances, it can transform into hormonally active disease presenting with severe Cushing syndrome. It might often produce diagnostic dilemma with difficult management issue if not thoroughly investigated and subtyped accordingly following surgery. Here, we present a 21-year-old male who initially underwent pituitary adenomectomy for presumed NFA with compressive symptoms. However, he developed recurrent and invasive macroadenoma with severe clinical as well as biochemical hypercortisolism during post-surgical follow-up. Repeat pituitary surgery was carried out urgently as there was significant optic chiasmal compression. Immunohistochemical analysis of the tumor tissue obtained on repeat surgery proved it to be an aggressive corticotroph adenoma. Though not cured, he showed marked clinical and biochemical improvement in the immediate postoperative period. Anticipating recurrence from the residual tumor, we referred him for cyber knife radio surgery. LEARNING POINTS: Pituitary NFA commonly present with compressive symptoms such as headache and blurred vision. Post-surgical development of Cushing syndrome in such a case could be either drug induced or endogenous. In the presence of recurrent pituitary tumor, ACTH-dependent Cushing syndrome indicates CD. Rarely a SCA presenting initially as NFA can transform into an active corticotroph adenoma. Immunohistochemical marker for ACTH in the resected tumor confirms the diagnosis.

Mutation Spectrum in TPO Gene of Bangladeshi Patients with Thyroid Dyshormonogenesis and Analysis of the Effects of Different Mutations on the Structural Features and Functions of TPO Protein through In Silico Approach.

Although thyroid dyshormonogenesis (TDH) accounts for 10-20% of congenital hypothyroidism (CH), the molecular etiology of TDH is unknown in Bangladesh. Thyroid peroxidase (TPO) is most frequently associated with TDH and the present study investigated the spectrum of TPO mutations in Bangladeshi patients and analyzed the effects of mutations on TPO protein structure through in silico approach. Sequencing-based analysis of TPO gene revealed four mutations in 36 diagnosed patients with TDH including three nonsynonymous mutations, namely, p.Ala373Ser, p.Ser398Thr, and p.Thr725Pro, and one synonymous mutation p.Pro715Pro. Homology modelling-based analysis of predicted structures of MPO-like domain (TPO142-738) and the full-length TPO protein (TPO1-933) revealed differences between mutant and wild type structures. Molecular docking studies were performed between predicted structures and heme. TPO1-933 predicted structure showed more reliable results in terms of interactions with the heme prosthetic group as the binding energies were -11.5 kcal/mol, -3.2 kcal/mol, -11.5 kcal/mol, and -7.9 kcal/mol for WT, p.Ala373Ser, p.Ser398Thr, and p.Thr725Pro, respectively, implying that p.Ala373Ser and p.Thr725Pro mutations were more damaging than p.Ser398Thr. However, for the TPO142-738 predicted structures, the binding energies were -11.9 kcal/mol, -10.8 kcal/mol, -2.5 kcal/mol, and -5.3 kcal/mol for the wild type protein, mutant proteins with p.Ala373Ser, p.Ser398Thr, and p.Thr725Pro substitutions, respectively. However, when the interactions between the crucial residues including residues His239, Arg396, Glu399, and His494 of TPO protein and heme were taken into consideration using both TPO1-933 and TPO142-738 predicted structures, it appeared that p.Ala373Ser and p.Thr725Pro could affect the interactions more severely than the p.Ser398Thr. Validation of the molecular docking results was performed by computer simulation in terms of quantum mechanics/molecular mechanics (QM/MM) and molecular dynamics (MD) simulation. In conclusion, the substitutions mutations, namely, p.Ala373Ser, p.Ser398Thr, and p.Thr725Pro, had been involved in Bangladeshi patients with TDH and molecular docking-based study revealed that these mutations had damaging effect on the TPO protein activity.

A rare variety of congenital adrenal hyperplasia with mosaic Klinefelter syndrome: a unique combination presenting with ambiguous genitalia and sexual precocity

Congenital adrenal hyperplasia (CAH) due to the three-beta-hydroxysteroid-dehydrogenase (3ß-HSD) enzyme deficiency is a rare autosomal recessive disorder presenting with sexual precocity in a phenotypic male. Klinefelter syndrome (KS) is the most common sex chromosome aneuploidy presenting with hypergonadotropic hypogonadism in a male. However, only a handful of cases of mosaic KS have been described in the literature. The co-existence of mosaic KS with CAH due to 3ß-HSD enzyme deficiency portrays a unique diagnostic paradox where features of gonadal androgen deficiency are masked by simultaneous adrenal androgen excess. Here, we report a 7-year-old phenotypic male boy who, at birth presented with ambiguous genitalia, probably a microphallus with penoscrotal hypospadias. Later on, he developed accelerated growth with advanced bone age, premature pubarche, phallic enlargement and hyperpigmentation. Biochemically, the patient was proven to have CAH due to 3ß-HSD deficiency. However, the co-existence of bilateral cryptorchidism made us to consider the possibility of hypogonadism as well, and it was further explained by concurrent existence of mosaic KS (47,XXY/46,XX). He was started on glucocorticoid and mineralocorticoid replacement and underwent right-sided orchidopexy on a later date. He showed significant clinical and biochemical improvement on subsequent follow-up. However, the declining value of serum testosterone was accompanied by rising level of FSH thereby unmasking hypergonadotropic hypogonadism due to mosaic KS. In future, we are planning to place him on androgen replacement as well. Learning points: •• Ambiguous genitalia with subsequent development of sexual precocity in a phenotypic male points towards some unusual varieties of CAH. •• High level of serum testosterone, adrenal androgen, plasma ACTH and low basal cortisol are proof of CAH, whereas elevated level of 17-OH pregnenolone is biochemical marker of 3ß-HSD enzyme deficiency. •• Final diagnosis can be obtained with sequencing of HSD3B2 gene showing various mutations. •• Presence of bilateral cryptorchidism in such a patient may be due to underlying hypogonadism. •• Karyotyping in such patient may rarely show mosaic KS (47,XXY/46,XX) and there might be unmasking of hypergonadotropic hypogonadism resulting from adrenal androgen suppression from glucocorticoid treatment.

Thyroid Dysfunction and Autoimmunity in First Trimester of Pregnancy, Single Center Experience in Bangladesh.

Study on thyroid function and thyroid autoantibody status in pregnancy considering the trimester specific range, is scarce in Bangladesh. This cross sectional study done in Bangabandhu Sheikh Mujib Medical University, Bangladesh from August 2012 to June 2013 encompassed 186 pregnant women of first trimester for study of thyroid function (TSH, FT4) and anti-thyroid antibodies (anti-TPO, anti-TG). Age of the subjects was 25.4±4.9 years (mean±SD), median gestational age was 9 weeks. Applying the trimester-specific normal reference range set by American Thyroid Association (ATA), 48(25.8%) of the women were found to have thyroid dysfunction; 40(21.5%) subclinical hypothyroid (SCH), 1(0.5%) overt hypothyroid (OH) and 7(3.8%) hyperthyroid; 40(21.5%) women had goiter. If non-pregnant adult normal rage is used, 22 of SCH women as per ATA criteria will be labeled as normal and 19 normal women as per ATA cut off will be labeled as hyperthyroid. There was statistically significant disparity for functional status defined by these two references cut off value (p<0.001). 29(15.6%) women had thyroid autoimmunity and the autoantibody positivity was more frequent in women with thyroid dysfunction than euthyroid women (22.92% vs. 13.04%, p<0.001). Even though universal screening for thyroid dysfunction is not yet a recommendation, it should be considered in our population.

Testing blood glucose may be useful in the management of dengue.

Recently dengue viral fever is observed each year in Bangladesh. Overall skills for diagnosis and management have improved owing to national awareness for the disease. We have observed and investigated the frequency of glucose intolerance in the early phase of dengue fever. A two-sampled challenge test by 75 gm oral glucose (OGTT) was done in 133 patients [age (mean+/-SD):33+/-13 years, sex (male/female): 97/36] suffering from dengue fever during their illness. Diagnosis of dengue was based on serologic test for anti-dengue antibodies after the first week. Other investigations were done as part of the management. Among 133 studied dengue patients, 100 were found to have glucose intolerance by OGTT (75.2%). In regards to intensity of intolerance, 21.1%(28/133) had diabetes while 54.1%(72/133) had impaired glucose tolerance (IGT). Among the patients who agreed for a second OGTT (n=40) during discharge, 11(28%) had normal OGTT at both events, 22(55%) revert to normal on second OGTT while 7(17.5%) persisted abnormal glucose intolerance. On regression analysis, glucose intolerance was independently related to increased age (p=0.001) and higher titre of IgG antibody (p=0.006). The study demonstrated that glucose intolerance is frequently associated with dengue fever in its early course. These findings may help for the early diagnosis of dengue fever; and warrants for avoidance of dextrose infusions as fluid replacement in dengue fever. Moreover, patients suffering from dengue fever should be cautioned for development of diabetes in future.

Endoscopic para-thyroidectomy: a new approach.

Two patients (one male and one female) underwent endoscopic para-thyroidectomy for parathyroid adenoma at the Department of Surgery, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh. Three ports (one mid line and two laterals) were employed, a 10 mm telescope was used for the visualization and a harmonic scalpel was used for the dissection. To the best of our knowledge, there was no report of endoscopic para-thyroidectomy from Bangladesh. Both patients were fed on the first post operative day and discharged from the 4th and 8th operative day. Both patient's parathyroid hormone (PTH) level dropped to about one fourth the level in 12 to 20 minutes after enucleation (as compared to the immediate pre operative level). Endoscopic para-thyroidectomy appears to be a technically feasible patient friendly modality of treatment for the selected cases of para-thyroid pathology in experienced hand with excellent cosmetic outcome.

Urinary iodine status and thyroid dysfunction: a Bangladesh perspective.

Iodine deficiency is endemic in Bangladesh. Compulsory iodization of table salt was introduced since 1993 to prevent and improve thyroid disorders in the country. Urinary iodine status, thyroid function and antithyroid antibodies were studied in 397 newly diagnosed thyroid patients and 94 age-sex matched controls. Among thyroid patients, 96 were hyperthyroid, 185 euthyroid and 116 hypothyroid. Mean and median urinary iodine were higher (p=0.075) in thyroid patients (26.13+/-0.91 and 23.03) than controls (22.65+/-1.47 and 18.59); in hyperthyroid and euthyroid than hypothyroid (p=0.020); in multinodular (28.08+/-2.80 and 26.94) and diffuse (27.35+/-1.19 and 26.71) goitre than uninodular (23.91+/-2.37 and 19.14) and nongoitrous (NG, 21.5+/-2.05 and 18.27) (p=0.098) patients but no sex difference (p=0.466). Antimicrosomal (26.7%) and antithyroglobulin (34%) antibodies were more frequently positive among thyroid patients than controls (6.4% and 12.8% respectively) (p=0.00002 and p=0.00005 respectively). Antibody positivity was higher in diffuse (82/228) and multinodular (20/47) goitre than nongoitrous (20/56) and uninodular (13/66) goitre (p=0.046) as well as in hypothyroid (55.2%) and hyperthyroid (36.5%) than euthyroid (19.5%) patients (P<0.001). Urinary iodine correlated neither with antimicrosomal (thyroid patients: p=0.597 and control: p=0.112) nor with antithyroglobulin (thyroid patients: p=0.388 and control: p=0.195) antibody. Thyroid autoimmunity and dysfunction seems common; and interaction of salt iodization with iodine status and thyroid disorders may be important in Bangladesh.

Chronic carriers of hepatitis B virus in Bangladesh: a comparative analysis of HBV-DNA, HBeAg/anti-HBe, and liver function tests.

Serological markers of hepatitis B virus (HBV), liver function tests and quantitative estimation of HBV-DNA are important in the assessment of the state of infection and prognosis following treatment for hepatitis B. This study aimed to determine whether low-cost assays, eg hepatitis B e antigen (HBeAg) and liver function tests, could be used for the assessment of infectivity as an alternative to HBV-DNA estimation. We tested 125 hepatitis B carriers for HBeAg, antibody to HBeAg (anti-HBe), and serum HBV-DNA; we also carried out a range of standard liver function tests. Seventy-three subjects were positive and 52 were negative for HBeAg. Of the HBeAg positive cases, 3 were also positive for anti-HBe; of the HBeAg negative cases, 5 were also negative for anti-HBe. Of these 8 cases, 7 had no detectable HBV-DNA. Most of the HBeAg positive but anti-HBe negative subjects were positive for HBV-DNA (74.3%; 52/ 70) whereas most of the HBeAg negative and anti-HBe positive subjects (93.6%; 44/47) were also negative for HBV-DNA. Of 56 HBV-DNA positive individuals, alanine transaminase (ALT) was found to be raised in 69.6% (p=0.066) and aspartate transaminase (AST) was raised in 66.1% (p=0.011), while 67.9% had normal alkaline phosphatase (ALP) (p=0.054). HBeAg (p=0.018) and raised ALT (p=0.008) were found to be independent predictors for HBV-DNA positivity among HBV carriers. This study suggests that HBeAg positive and anti-HBe negative hepatitis B carriers with raised ALT and AST are likely to be positive for HBV-DNA; the combination of routine serology and biochemical tests may be considered as an alternative to HBV-DNA in evaluating the state of chronic HBV infection. However, HBV-DNA should be specifically assessed if discordance is observed between seromarkers and transaminases.

Predominance of the DEN-3 genotype during the recent dengue outbreak in Bangladesh.

A recent outbreak of dengue in Bangladesh was marked by many fatal complications. As clinical virulence varies among the genotypes of dengue virus, a study was conducted to investigate the molecular genotypes of dengue in Bangladesh. Reverse transcription polymerase chain reaction was used to determine viral genotypes using oligonucleotide generic primers that produce a 511 bp product. The resulting product was typed by nested PCR with strain-specific primers, yielding 482 (DEN-1), 119 (DEN-2), 290 (DEN-3) and 392 (DEN-4), visualized on UV transilluminator after electrophoresis on 2% agarose gel stained with ethidium bromide. Of 45 clinically diagnosed dengue patients (mean age 28 years; male/female 30/15), 19 (42.2%) had detectable viral RNA in their blood. However, during the first 5 days of fever in 30 patients, the frequency was 60% (18/30), implying that the sooner serum is drawn after the fever, the greater the chances of detecting viral RNA. DEN-3 was detected in all except 2 patients who were infected with DEN-2. DEN-2 (two cases) and DEN-4 (one case) were present as co-infections with DEN-3. All of the patients presented with fever, anorexia and vomiting; many had headache and general body ache; a few had a rash. About a quarter had suffered episodes of bleeding, while ascites, pleural effusion and CNS symptoms were found in a few patients Patients positive for viral RNA were also positive for anti-dengue IgM (p=0.007) in subsequent sampling. The study suggests the predominance of DEN-3 infection with occasional co-infection with other types, during the recent outbreak of dengue in Bangladesh.

Status of antithyroid antibodies in Bangladesh.

To study autoimmunity among thyroid diseases, 397 thyroid patients (age 30 (13) years; M/F 75/322) from two referral centres in Bangladesh and 94 healthy controls (age 30 (13) years; M/F 24/70) were studied for antimicrosomal and antithyroglobulin antibodies. Thyroid patients were clinically grouped as suspected autoimmune thyroid disease (AITD), non-autoimmune, or indeterminate groups (where no decision could be reached). Antimicrosomal antibody was strongly positive in 19.4% and weakly positive in 7.3% of patients but only 4.3% and 2.1% respectively in the controls (chi(2) = 17.852; p = 0.000) whereas strong and weak positivity were 27.2% and 6. 8% in patients compared with 8.5% and 4.3% respectively in the controls (chi(2) = 16.916; p = 0.000) for antithyroglobulin antibody. Antibodies were positive in 63.0% with Hashimoto's thyroiditis, 36.4% with Graves' disease, and 44.7% with atrophic thyroiditis among the autoimmune group. In the non-autoimmune group antibodies were positive in 100% with multinodular hypothyroidism, 46.7% with subacute thyroiditis, 40.0% with suspected iodine deficiency goitre, 31.3% with toxic multinodular goitre, 30.8% with non-toxic solitary nodules, and 19.4% with simple diffuse goitre. None was positive for antimicrosomal antibody without being positive for antithyroglobulin antibody. The two antibodies strongly correlated in both patients (r = 0.977, p = 0.000) and controls (r = 0.986, p = 0.000). About 9% (36/397) of patients were mismatched with the final diagnosis on antibody measurement; most of them had Hashimoto's thyroiditis (33/36). Prevalence of AITD among thyroid patients was 48.36%. Specificity of antimicrosomal and antithyroglobulin antibodies were 93% and 87%. It was concluded that AITD is not uncommon in Bangladesh; antimicrosomal antibody is a useful marker for AITD and unless antibodies are checked, an appreciable number of patients with AITDs will remain undetected.