Rapid culture diagnosis of tuberculous lymphadenitis from a tertiary care centre in an endemic nation: Potential and pitfalls.

In spite of low sensitivity and specificity, standard diagnostic algorithm recommends fine needle aspiration cytology (FNAC) and direct microscopic screening for acid-fast bacilli (AFB) for the routine diagnosis of tuberculous lymphadenopathy (LNTB). In this study, the diagnostic utility of liquid broth based automated culture (BacT/ALERT 3D) technique was assessed in comparison with conventional techniques in 89 clinically suspected tubercular lymphadenitis patients. 60% (n = 53) were positive by FNAC and 38.4% (n = 34) demonstrated AFB in smear examination. BacT/ALERT yielded isolation in 43.1% (n = 38) aspirates, confirming tubercular aetiology. We also found six paediatric culture-positive cases which showed negative outcome by both FNAC and smear. Thus, we conclude that culture by BacT/ALERT, may be used for faster yield of Mycobacteria in LNTB, especially in children. Additionally, this could also be used as a platform for further differentiation of Mycobacterium tuberculosis from non-tuberculous mycobacteria (NTM) infection and for testing of anti-tubercular chemotherapeutic agents whenever drug resistance is suspected.

Assessment of trends of ofloxacin resistance in Mycobacterium tuberculosis.

Purpose: Ofloxacin (OFX) is one of the potent fluoroquinolone (FQ) recommended to treat MDR-TB. Over a decade, the preexposure of this drug for the treatment of other bacterial infections has resulted in acquisition of FQ resistance among Mycobacterium tuberculosis strains. Considering this possibility, a study was undertaken in a tertiary care center in the capital city (India) to assess the drug resistance trends of OFX among susceptible and multidrug resistant (MDR) strains of M. tuberculosis. Materials and Methods: A total of 102 M. tuberculosis isolates (47 susceptible to first-line drugs and 55 MDR isolates) were screened for susceptibility testing of OFX with a critical concentration of 2 μg/ml by Lowenstein Jensen (LJ) proportion method. Results: The results showed 40 (85.1%) isolates among 47 susceptible isolates and 34 (61.8%) isolates among 55 MDR isolates, were found to be susceptible to OFX. Fisher's exact test showed significant P-value (0.0136) demonstrating 1.377 fold (95% confidence interval) increased risk to become resistant to OFX than susceptible isolates. These finding shows decreased OFX susceptibility is not only limited to MDR isolates but also increasingly seen in susceptible strains as a result of drug abuse. Conclusions: Our finding were not alarming, but highlights the general risk of acquiring resistance to OFX, jeopardizing the potential for these drugs to be used as second-line anti-TB agents in the management of drug-resistant TB and creating incurable TB strains.

Evaluation of Nucleic Acid Probe for Rapid Identification of Mycobacterium tuberculosis Complex in Extra-Pulmonary Culture Isolates.

Patients infected with Non-tuberculous mycobacteria (NTM) usually do not respond to conventional anti-tubercular treatment and are misdiagnosed as infection with multi-drug resistant strains of mycobacterium tuberculosis (M.tb) due to lack of correct species identification, particularly in the developing countries like India. One of the challenges faced by clinicians in the treatment of tuberculosis is the absence of an easy, reliable and rapid identification tool that can accurately differentiate disease caused by M.tb complex from NTM. Keeping this in consideration, the performance of species specific nucleic acid probe i.e. Accuprobe was assessed and compared with conventional niacin production, nitrate reductase assay techniques for identification of M.tb complex in 80 mycobacterial isolates obtained from different extra - pulmonary sites. Accuprobe identified 62 isolates (77.5%) as M. tuberculosis complex and remaining 18 isolates (22.5%) as NTM whereas 64 isolates (80%) were identified as M.tb and rest 16 (20%) were interpreted as NTM by conventional biochemical techniques. The overall agreement between both techniques was 96.9% The sensitivity, specificity, positive predictive value(PPV) and negative predictive value (NPV) shown by accuprobe were 96.9%, 100%, 96.9%, and 88.9% respectively. Thus, accuprobe has showed impressive sensitivity and specificity giving results in <3 hrs from culture-positive isolates and have sure edge over conventional biochemical methods which are, nonetheless, labour intensive and cumbersome to perform thus delaying prompt mycobacterial identification.

The use of E-test for the drug susceptibility testing of Mycobacterium tuberculosis - A solution or an illusion.

Aim: To evaluate E-test as a tool for rapid determination of drug susceptibility against the conventional LJ method focusing on reliability, expense, ease of standardization and performance of the technique in low resource settings. Materials and Methods: A total of 74 clinical isolates (2004-2005) of Mycobacterium tuberculosis were tested using E-test for susceptibility to streptomycin (STM), isoniazid (INH), rifampicin (RIF) and ethambutol (EMB) by E-strip and LJ (LJPM) proportion methods. Results: The LJPM method, the gold standard, detected resistance against STM in 16.2%, INH in 40.5%, RIF in 18.9% and EMB in 27% cases. In comparison, the resistance values showed by E-test was 66.67% for STM, 57.14% for INH 71.43% for RIF and 80% for EMB. The susceptible correlation was 90.32% for STM, 73.91% for INH, 93.33% for RIF and 59.26% for EMB. E-test correctly identified only eight of the 12 (66.6%) MDR isolates and wrongly identified four isolates which were not MDR. The overall agreement between the two methods was only 48.6%. Resistant isolates showed false positive resistance observed while using E-strip towards all the drugs. Conclusion: E-strips are not quite feasible as a replacement for LJ-proportion method on a large scale due to high risk of cross contamination, laboratory infection, expense associated with it and high false positive resistance observed to all first line drugs. However, the good correlation observed for RIF between the two methods indicates that E-test could contribute to the role in rapid screening of MDR TB isolates as rifampicin mutations are invariably observed in MDR TB isolates.

Paraoxonase (PON1) activity in north west Indian Punjabis with coronary artery disease & type 2 diabetes mellitus.

BACKGROUND & OBJECTIVE: Paraoxonase (PON1), an arylesterase is associated with high density lipoprotein cholesterol (HDL-C). PON1 prevents low density lipo-protein cholesterol (LDL-C) from peroxidation and can also hydrolyze lipid peroxides, thereby providing protection against atherosclerosis and coronary artery disease (CAD). The incidence of CAD is known to be high in north western Indian Punjabis. Though many factors may play a role in its pathogenesis, low PON1 activity could be an independent risk factor. We carried out this study to determine PON1 activity in north-west Indian Punjabi patients with CAD with and without type 2 diabetes mellitus and compared with healthy individuals. METHODS: A total of 120 patients with angiographically proven CAD (57 with and 63 without type II diabetes mellitus) and 19 healthy controls were studied for plasma PON1 activity and lipid variables. Comparison was undertaken between CAD patients and healthy controls and between CAD patients with and without type II DM. RESULTS: Significantly lower plasma PON1 activity (P<0.05) along with lower HDL-C (P<0.001) and higher LDL-C (P<0.05) levels were observed in CAD patients as compared to healthy controls. On univariate analysis of variance after adjusting for age and sex, no significant difference could be observed between PON1 activity and age and sex. On discriminant analysis, no clear cut-off could be observed in PON1 activity between patients CAD and controls. Similarly between CAD with and without patients type II diabetes mellitus, there was no significant difference in PON1 activity and lipids. INTERPRETATION & CONCLUSION: The low plasma PON1 activity irrespective of being diabetic may be an independent risk factor for CAD in north-western Indian Punjabi population. Similar studies involving larger samples in different ethnic groups in India need to be done to find out the role of PON1 activity in CAD.

Noninvasive estimation of clinically asymptomatic pulmonary hypertension in idiopathic pulmonary fibrosis.

OBJECTIVES: To determine the prevalence of pulmonary hypertension in patients with idiopathic pulmonary fibrosis (IPF) by noninvasive methods. PATIENTS AND METHODS: Twenty seven patients with IPF attending the Chest Clinic over a period of one-and-a-half-years underwent echocardiography for evidence of pulmonary hypertension, which was defined as pulmonary artery systolic pressure > or = 40 mmHg by Doppler echocardiography, or pulmonary acceleration time < or = 100 milliseconds or two-dimensional echocardiographic findings of right ventricular hypertrophy or overload. RESULTS: Two patients with clinical evidence of pulmonary hypertension were excluded from analysis. Their mean age (n = 25) was 53.8 years. The mean duration of symptoms before presentation was 2.1 years. Pulmonary hypertension was diagnosed by echocardiography in nine of the 25 patients (36%). There was a statistically significant difference between the duration of illness, arterial oxygen tension (PaO2) levels and forced vital capacity in patients with pulmonary hypertension than in those without pulmonary hypertension. Logistic regression analysis showed a significant association between the duration of illness, decreasing forced vital capacity and hypoxemia with the development of pulmonary hypertension. CONCLUSIONS: Echocardiography detects pulmonary hypertension in clinically asymptomatic individuals, and should be used routinely for the diagnosis of pulmonary hypertension in patients with IPF.

Expression and functional activity of receptor-Ck in mononuclear cells of a homozygous hypercholesterolemic family.

The study was addressed to explore the expression and functional activity of a novel cholesterol-specific cell surface receptor-Ck in a typical homozygous familial hypercholesterolemic family. Functional activity of receptor-Ck was characterized by its ability to downregulate Bcl-2 gene expression through a 47 kDa factor having an affinity for the sterol-regulatory element in the promoter region of this gene. The result of such a study revealed normal expression and functional activity of receptor-Ck accompanied by a lack of Apolipoprotein B-specific low-density lipoprotein receptor gene expression in the mononuclear cells derived from these patients. On the basis of these results, it is tempting to speculate that receptor-Ck may be involved in the maintenance of cellular cholesterol homeostasis observed in homozygous familial hypercholesterolemic patients.

Perioperative myocardial infarction in coronary artery disease patients and 'at-risk' for coronary artery disease patients undergoing non-cardiac surgery.

BACKGROUND: Perioperative myocardial infarction (POMI) carries a high mortality and occurs more commonly in patients with a history of coronary artery disease (CAD). However, there are also other patients undergoing surgery who are 'at risk' for CAD but who do not have a history of infarction or angina. We compared the incidence of POMI in these two groups. METHODS: In a prospective study of 69 men and 39 women over 30 years of age undergoing non-cardiac surgery under general or regional anaesthesia, 56 had definite CAD and 52 were 'at-risk' for CAD. All these patients were followed up with serial postoperative electrocardiography and CK-MB isoenzyme analysis for the diagnosis of POMI. RESULTS: The POMI rate was 32% in definite CAD patients and 15% in patients 'at-risk' for CAD. Mortality in patients with POMI was 17% in those with CAD and 13% in those 'at-risk' for CAD. Perioperative myocardial infarction was maximal in the first 24 hours following surgery (77%). All the POMIs were painless. Anaesthesia techniques--whether regional or general--did not influence the incidence of POMI (Chi-square, p > 0.05). The type of drugs used in the treatment of CAD such as beta-blockers, calcium channel blockers and antiplatelet agents did not cause any difference in the incidence of POMI (Chi-square, p > 0.05). Patients who had either an intraoperative hypertensive episode, tachycardia, arrhythmias or ST-segment changes had a higher incidence of POMI (Chi-square, p > 0.05). The incidence of POMI was not lower in patients undergoing transurethral resection of the prostate compared to patients undergoing other types of non-cardiac surgery (Chi-square, p > 0.05). CONCLUSION: POMI occurs in one-third of patients with a history of CAD and one-sixth of those 'at-risk'. It carries a mortality of 17% and 13% respectively. Decisions to operate on such patients should be taken with caution.