Rapid and sensitive detection of major uropathogens in a single-pot multiplex PCR assay.

Urinary tract infection (UTI) is among the most common bacterial infections and poses a significant healthcare burden. Escherichia coli is the most common cause of UTI accounting for up to 70 % and a variable contribution from Proteus mirabilis, Pseudomonas aeruginosa and Klebsiella pneumoniae. To establish a complete diagnostic system, we have developed a single-tube multiplex PCR assay (mPCR) for the detection of the above-mentioned four major uropathogens. The sensitivity of the assay was found to be as low as 10(2) cfu/ml of cells. The mPCR evaluated on 280 clinical isolates detected 100 % of E. coli, P. aeruginosa, P. mirabilis and 95 % of K. pneumonia. The assay was performed on 50 urine samples and found to be specific and sensitive for clinical diagnosis. In addition, the mPCR was also validated on spiked urine samples using 40 clinical isolates to demonstrate its application under different strain used in this assay. In total, mPCR reported here is a rapid and simple screening tool that can compete with conventional biochemical-based screening assays that may require 2-3 days for detection.

Cutaneous cryptococcosis among HIV infected patients.

Five patients with HIV associated cutaneous cryptococcal infection are reported.

Cryptococcal meningitis among HIV infected patients.

Cryptococcal meningitis is an emerging opportunistic infection among HIV infected patients and an important cause of mortality among these patients. The incidence of cryptococcal meningitis varies from place to place. A total of 31 specimens of CSF out of 89 samples processed from known HIV positive cases yielded Cryptococcus neoformans during the period of 3 years. C.neoformans was the most common opportunistic pathogen isolated from CSF samples of these patients with an incidence of 34.8%

Amiodarone--induced changes in surfactant phospholipids of rat lung.

Amiodarone HCl (AD) is a very effective antiarrhythmic drug, but its use is often associated with serious pulmonary complications. It is shown to induce lung phospholipidosis. Nevertheless, the effects of this drug on pulmonary surfactant which is composed of about 75% phospholipids and which prevents alveolar collapse is not known. Therefore, we have examined the effect of AD on the intra- and extracellular surfactant pools and on the levels of phosphatidylcholine (PC), the primary constituent of pulmonary surfactant. Male Wistar rats were fed AD (175 mg/kg) by oral gavage for three weeks. At the end of the experimental period, the rats were killed, the lungs removed and perfused, and surfactant isolated. Some lungs were prepared for ultrastructural examination. Phospholipid was assayed in the intra- and extracellular surfactant. Amiodarone produced a significant increase in both the intra- and extracellular surfactant phospholipid along with an appreciable change in the phospholipid profile. Also, the drug seemed to increase the number of lamellar inclusions in the surfactant producing type II alveolar cells. These data suggest that administration of AD leads to an increase in the lung surfactant phospholipid levels and lamellar bodies in alveolar type II cells.

Effect of amiodarone, an antiarrhythmic drug, on serum and liver lipids and serum marker enzymes in rats.

Administration of amiodarone (AD) to rats leads to marked damage to liver, as evidenced by pathological changes and significant increases in activities of serum marker enzymes and levels of lipids like cholesterol and phospholipids with no alteration in the triglyceride levels. The risk factor, that is the total cholesterol/HDL cholesterol ratio, exhibited increase in the experimental animals, indicating that amiodarone treatment may lead to the development of coronary heart disease.

Haematological and erythrocyte membrane changes induced by amiodarone, in rats.

The effect of Amiodarone (AD), a cationic amphiphilic drug, on erythrocytes and leucocytes was studied. Treatment of rats with AD showed a significant decrease in the red cell count and the level of Hemoglobin. Amiodarone altered the fluidity of the erythrocyte membrane followed by a decrease in the activities of membrane bound enzymes like (Na+, K+)-ATPase, Acetylcholine esterase and NADH dehydrogenase. A slight increase in the leucocyte count was also observed in the treated animals.

Lung injury by amiodarone, an antiarrhythmic drug, in male rats.

Administration of single dose (175 mg/kg body wt) of amiodarone dissolved in water through gavage for 3 weeks damaged the lung and changed the content of lung lavage. Activities of bronchoalveolar lavage (BAL) angiotensin converting enzyme (ACE) and lactate dehydrogenase (LDH) increased significantly. Also, the protein and lactate content of the lavage fluid increased significantly over the control. The drug also produced marked changes in morphology of the lung of experimental animals.