Extraction and detection of Mycobacterium leprae DNA from ZNCF-stained skin smear slides for better identification of negative skin smears.

Identification of Mycobacterium leprae, which causes leprosy, is done by Ziehl Neelsen Carbol Fuchsin (ZNCF) stained slit skin smear microscopy that aids in the diagnosis and quantification of approximate bacterial load carried by the patient. We attempted M. leprae DNA extraction from 46 stained slit skin smear negative slides, using Proteinase K and SDS lysis, followed by ethanol precipitation. M. leprae specific primers (16SrRNA) were used for PCR-based amplification of DNA. We could detect M. leprae DNA in 15 (32.6%) samples. The method can be useful in the diagnosis of apparently slit skin smear negative leprosy cases.

Use of reverse transcription polymerase chain reaction for the detection of Mycobacterium leprae in the slit-skin smears of leprosy patients.

The relevance of bacterial index (BI) for understanding the prognosis of leprosy patients on treatment has been extensively debated, as it does not give a very clear idea of the viability of the bacteria in patients under treatment. Here we used slit-skin smear samples of leprosy patients to test the suitability for studying viability of Mycobacterium leprae using reverse transcription polymerase chain reaction (RT-PCR). For this purpose, we recruited 13 multibacillary (MB) leprosy patients (8 lepromatous and 5 borderline lepromatous). Of these, 7 were relapse cases, 3 were under treatment (MB-MDT), 2 were new cases and 1 had completed treatment. We carried out extraction of RNA using Trizol reagent (Life Technologies, UK) from the slit-skin smear samples from these patients. The RNA preparation was then used for the RT-PCR using Mycobacterium leprae-specific primers for the fragment of 16s ribosomal RNA gene. Samples from both the new cases, 4 suspected relapse cases and 1 patient under treatment showed positive RT-PCR results. Other 6 patients whose smear samples did not show any amplification by RT-PCR were on MB-MDT from 8 to 30 months. The usefulness of the technique needs to be validated using mouse footpad technique and also should be more extensively explored for studying the viability of M. leprae, the efficacy of treatment and the presence of other mycobacteria in the slit-skin smear samples.

Transdiaphragmatic extension of hepatic hydatid cyst.

Transdiaphragmatic extension of hydatid cyst (HC) or cystic echinococcosis (CE) of the liver is a rare phenomenon. We report a case that presented as a right middle lobe consolidation. The diagnosis of transdiaphragmatic extension of hepatic hydatid cyst was suspected on CT scan of the chest and abdomen, and confirmed operatively. A successful outcome was achieved by a combination of pre- and post-operative albendazole therapy combined with surgery.

Non responding pneumonia with skin lesions.

Sweet's syndrome (acute febrile neutrophilic dermatitis) is characterised by classical skin lesions accompanied by fever and malaise. Systemic involvement may be present and lung involvement in Sweet's syndrome has been reported in the form of bilateral pulmonary infiltrates, bronchiolitis obliterans organising pneumonia and pleural effusion. There are dense papillary neutrophilic infiltrates on histopathology. We present a case of Sweets' syndrome with left lower lobe consolidation and persistent fever which was non-responsive to antibiotics but showed clinical improvement with clearing of radiological opacities on oral steroid therapy.

Urinothorax: a case report.

A case of right sided transudative pleural effusion associated with right sided renal calculi and moderate hydronephrosis is reported. The diagnosis of urinothorax was confirmed by demonstrating a pleural fluid to serum creatinine ratio of greater than one. The relief of obstructive uropathy by surgical treatment of renal calculi resulted in resolution of the pleural effusion.

Recurrent respiratory papillomatosis.

Recurrent respiratory papillomatosis is characterized by the appearance of benign laryngeal squamous papillomas in childhood. Lung involvement is rare. We report a case of childhood laryngeal papillomas who developed tracheobronchial papillomas and a nodule in the lung after a period of 21 years. Frequent sampling of pulmonary lesions to detect malignant transformation is suggested as prognosis of lung lesions are worse in comparison to laryngeal papillomas.

Blood stem cell transplantation: current concepts.

Mobilized peripheral blood haematopoietic progenitor cells are increasingly being used as against bone marrow (BM) transplants, following high dose chemotherapy and/or radiotherapy for the management of chemosensitive malignancies. Rapid haematopoietic reconstitution as evidenced by reduced duration of neutropaenia, fewer donor platelet infusions, shorter hospital stay and reduced cost of treatment are the advantages of this procedure. Reduced tumour cell contamination of mobilized blood compared to bone marrow however, has not been substantiated. Mobilization of lymphokine activated killer cells (LAK), use of blood stem cells (BSC) for allogeneic transplants and ex vivo expansion of the mobilized cells are emerging as the future areas for research. Addition of interleukin-3 (IL-3), stem cell factor (c-kit ligand) and PIXY-321 appear to open-up new vistas by enforcing trilineage and multilineage haematopoietic reconstitution.