Evaluation of nested Polymerase Chain Reaction targeting hup B gene in the diagnosis of tubercular ascites

Abdominal tuberculosis usually has nonspecific presentation, frequently mimicking other diseases. Because of the limitations of the conventional methods of diagnosis of extra pulmonary tuberculosis, focus is shifted to molecular methods. The objectives of this study are to evaluate the role of nested Polymerase Chain Reaction (PCR) targeting hup B gene as a rapid diagnostic modality of tubercular ascites and also to detect the infecting species (Mycobacterium tuberculosis and Mycobacterium bovis). 100 suspected tubercular ascites patients were enrolled in the study. Ascitic fluid was processed by Universal Sample Processing (USP) method and two steps nested PCR was performed targeting hup B gene. Patients were put on Anti Tubercular Therapy [Category I, (2 HRZE + 4 HR) 3, RNTCP, India]. A positive response to therapy was considered as gold standard and PCR assay was compared to determine sensitivity, specificity, positive and negative predictive value in diagnosis of tubercular ascites. 79 patients could be followed up to see the response to therapy. Of these, 39 were PCR positive and 35 responded to Anti Tubercular therapy. The sensitivity, specificity, positive and negative predictive value were found to be 97.1%, 88.6%, 87.2% and 97.5% respectively. The good sensitivity and specificity obtained in the study suggests the use of PCR targeting hup B gene as a routine diagnostic tool for tubercular ascites. Another added advantage is the ability to identify between M. tuberculosis and M. bovis which otherwise have similar clinical presentation.Keywords: Hup B gene; M. tuberculosis; M. bovis; Nested Polymerase Chain ReactionInternet Journal of Medical Update 2012 July;7(2):9-1

Urban-Rural Differences in Atherogenic Dyslipidaemia (URDAD Study): A Retrospective Report on Diabetic and Non-diabetic Subjects of Northern India

Diabetes and urbanization are major contributors to increased risk factors of cardiovascular diseases. Studying whether atherogenic dyslipidaemia increases with urbanization in type 2 diabetes mellitus is, therefore, important. The sample of the present study consisted of 400 subjects. They were categorized according to residential area and diabetes into four groups: urban diabetic group, urban non-diabetic control group (from a metropolitan city Delhi), rural non-diabetic diabetic group, and rural control group (from villages of Khanpur Kalan, Sonepat, Haryana). Differences in lipid levels and risk factors of emerging cardiovascular diseases between groups were evaluated with analysis of variance. Diabetic patients of both urban and rural areas had significantly higher total cholesterol (TC), triglycerides (TG), very low-density lipoproteins (VLDL), TC to high-density lipoprotein cholesterol (TC/HDL) ratio, TG to high\u2011density lipoprotein cholesterol (TG/HDL) ratio, and atherogenic index (AI) compared to respective controls (p<0.05). The HDL concentrations in urban diabetics were significantly lower (p<0.05) than in urban non-diabetic group and rural diabetic group. Comparison between urban and rural diabetic groups showed significantly higher atherogenic dyslipidaemia (AD) in the urban patient-group (p<0.05). We evaluated significant relationships of diabetes and urbanization with AD by multiple regression analysis. Receiver operating curve (ROC) analysis showed high area under curve (AUC) for TG/HDL in urban diabetic group (0.776, p<0.0001) and in rural diabetic group (0.692, p<0.0001). It is concluded that diabetes was associated with higher AD parameters. Urbanization in diabetes is also associated with elevated levels of AD, indicating higher risk in urban population. This study suggests that TG/HDL may be particularly useful as atherogenic risk predictor in newly-diagnosed type 2 diabetic patients

Biochemical markers of tubercular ascites

ABSTRACT The diagnosis of abdominal tuberculosis requires a high index of suspicion due to its vague symptomatology. Early diagnosis of tubercular ascitis is crucial to prevent progression of disease to its advanced stages, thereby preventing the fatal complication like intestinal obstruction, fistulas and peritonitis. The objective of our research work is to evaluate the role of biochemical parameters such as Adenosine Deaminase (ADA), IgG, Lactate, Total protein and albumin, Glucose, Cholesterol and pH in diagnosis of abdominal tuberculosis. Ascitic fluid samples were taken from patients admitted in medicine wards of SSK Hospital after informed consent. A total of 100 patients meeting the selection criteria were enrolled in the study. The biochemical investigations performed for the ascitic fluid samples were Adenosine Deaminase (ADA), IgG against 38 kDa mycobacterial antigen, Lactate, Total protein, albumin, Glucose, Cholesterol and pH. In the 79 patients that had been followed up, with ATT response as a reference, a highly significant association was observed with ascitic fluid assays ADA, IgG, Serum Ascitic Albumin Gradient(SAAG), Cholesterol, Lactate and pH. Adenosine Deaminase (ADA) and IgG against 38 kDa mycobacterial antigen can be used as corroborative markers for diagnosis of extra pulmonary paucibacillary tubercular ascitis where conventional methods like smear microscopy and culture frequently fail to establish the diagnosis

Emerging Trend of Mutation Profile of rpoB Gene in MDR Tuberculosis, North India

The present study was conducted on North Indian population to observe rpoB gene mutation profile in multidrug resistant Mycobacterium tuberculosis. This was an observational study. 30 cases of MDR-TB proven by culture and drug sensitivity were selected. DNA sequencing of 81 bp (codon 507–533) long RRDR of Mycobacterium tuberculosis was done to detect the sites of mutation. Out of 30 cases, 24 showed a single mutation in the RRDR region of rpoB gene in which 16 (53.33 %) showed mutation in codon 531(TCG→TTG), 5 cases (16.66 %) showed mutation in codon 526(CAC→TAC), mutation in codon 516(GAC→GTC, AAC) was present in 3 cases (10 %). It was also observed that mutation in more than one codon was present in 4 cases (13.33 %), which included deletion at codon 509(AGC→–GC), mutation at 513(CAA→CTA), 516, 526, 529(CGA→CTA) and 531. No mutation was detected in RRDR in 2 cases (6.66 %). Our finding of 13.33 % cases with multiple sites of mutation in RRDR region is in contrast to earlier studies done in North India which showed single mutation detected in RRDR of rpoB gene that highlights the emerging change in the trend of mutation profile of rpoB gene in rifampicin resistant Mycobacterium tuberculosis

Raised TSH is associated with endothelial dysfunction in Metabolic Syndrome: A case control study

Introduction. Endothelial dysfunction has been considered as one of the important factors in pathogenesis of Metabolic Syndrome (Met S). Subclinical hypothyroidism (SCH) has also been reported to be associated with Met S. The aim of our study is to evaluate the association of raised TSH with mediators of endothelial dysfunction in Met S with Subclinical hypothyroidism as compared to healthy controls

Raised TSH is associated with endothelial dysfunction in Metabolic Syndrome: A case control study

Abstract Introduction. Endothelial dysfunction has been considered as one of the important factors in pathogenesis of Metabolic Syndrome (Met S). Subclinical hypothyroidism (SCH) has also been reported to be associated with Met S. The aim of our study is to evaluate the association of raised TSH with mediators of endothelial dysfunction in Met S with Subclinical hypothyroidism as compared to healthy controls. Methods. Study population consisted of 100 subjects, out of which 50 were cases of Met S and 50 were healthy controls. Met S group were further divided into two, based on the presence &amp; absence of SCH. Serum insulin, T3, T4, TSH were measured by chemiluminescence based immunoassay (CLIA). Serum nitric oxide (NO) levels were measured by Modified Griess’s method and serum endothelin-1 (ET-1) levels were measured by ELISA. Results. Out of 50 cases of Met S, SCH was diagnosed in 22. The mean serum TSH levels were significantly higher in Met S cases as compared to healthy controls (5.7 ± 1.2 μIU/mL vs. 2.3 ± 1.6 μIU/mL, P &lt;0.0001). Mean serum NO levels were significantly lower in Met S cases as compared to healthy control (15.4 ± 10 μM vs. 21 ± 10 μM, p = 0.009). Mean serum ET-1 levels were significantly higher in Met S cases as compared to healthy controls (2.68 ± 1.7 fmol/mL vs. 2.1 ± 0.84 fmol/mL, p = 0.011). On Pearson’s correlation analysis, TSH showed positive correlation with ET-1 (r = 0.341, p = 0.001) and negative correlation with NO (r = −0.331, p = 0.001). Binary logistic regression analysis showed that TSH, NO and ET-1 has significant odd’s ratio for predicting Met S. Conclusion. Met S cases were screened for thyroid abnormalities and found to have 44% of SCH along with co-existing endothelial dysfunction. Raised TSH in SCH could cause endothelial dysfunction which may lead to Met S and associated co-morbidities. Present study gives new insight in linking endothelial dysfunction and raised TSH in Met S. Therefore, Met S cases should be screened for SCH and treated appropriately to attenuate endothelial dysfunction and associated comorbidities in Met S.</jats:p

Association of prothrombotic adipokine (plasminogen activator inhibitor-1) with TSH in metabolic syndrome: a case control study

Abstract Background Metabolic syndrome (MetS) involves a cluster of cardiovascular risk factors, including abnormal lipids, insulin resistance and hypertension. The aim of the present study is to investigate associations between thyroid profile and the pro-thrombotic mediator, plasminogen activator inhibitor-1 (PAI-1), in MetS and identify associated biochemical markers. Materials and methods The present study was a case control study and consisted of 50 diagnosed cases of MetS and 50 healthy volunteers as controls. MetS cases were further divided into two groups based on the presence and absence of subclinical hypothyroidism (SCH). Data collected included demographic profile, clinical history and routine lab investigation. Special investigations included the thyroid function test and serum PAI-1 levels. Results The mean serum thyroid-stimulating hormone (TSH) levels were significantly higher in MetS cases as compared to controls (5.7 ± 1.2 mIU/L vs. 2.3 ± 1.6 mIU/L, p &lt; 0.0001), although the mean triiodothyronine (T3) and thyroxine (T4) levels were comparable in two groups. The mean levels of serum PAI-1 were significantly higher in MetS cases as compared to controls(231 ± 87 ng/mL vs. 185 ± 96 ng/mL, p = 0.013). TSH and PAI-1 levels were positively correlated with various markers of MetS and negatively correlated with high-density lipoprotein (HDL). Conclusion The present study points towards the presence of thyroid dysfunction, in the form of subclinical hypothyroidism (SCH), in cases of MetS. In the presence of thyroid dysfunction, abnormal adipocytes may release adipokines, such as PAI-1, which lead to increased risk of thrombotic episodes in these patients. Hence, SCH should be appropriately managed. </jats:sec

Study of Oxidative Stress in Vitiligo

Vitiligo is an idiopathic, acquired, circumscribed, hypomelanotic skin disorder, characterized by milky white patches of different sizes and shapes. It is due to the destruction of melanocytes resulting in the absence of pigment production of the skin and mucosal surfaces. Oxidative stress has been implicated in pathophysiology of vitiligo. To study the activity of blood Superoxide dismutase (SOD) and Glutathione peroxidase (GPx) in vitiligo patients. A case–control study was conducted in which 100 patients were enrolled after written consent. 50 cases were of active vitiligo and 50 served as control (25 healthy control and 25 with stable vitiligo). SOD—In our study, among the active vitiligo cases 90% had high level of SOD and 10% had normal level of SOD. Among the stable vitiligo controls, 92% had normal level of SOD and 8% had low levels of SOD.The difference between active vitiligo cases and stable vitiligo control as well as with healthy control was statistically significant (P value < 0.05). GPx—Among the active vitiligo cases 74% had normal GPx levels, 22% had low and only 4% had high levels of GPx. Among the stable vitiligo controls, 64% had normal GPx levels, 16% had low, and 20% had high levels of GPx. The difference between active vitiligo cases and stable vitiligo control as well as with healthy control was statistically not significant (P value > 0.05). Our study shows that oxidative stress is involved in the pathophysiology of vitiligo, as indicated by the high levels of serum superoxide dismutase activity