Development of TaqMan real-time polymerase chain reaction for the detection of the newly emerging form of carbapenem resistance gene in clinical isolates of Escherichia coli, Klebsiella pneumoniae, and Acinetobacter baumannii.

Purpose: The newly emerging form of the so-called New Delhi Metallo-beta-lactamases (NDM-1) has been reported recently from patients worldwide and broadly thought as a potential source for the major global health problem. Thus, it is important to study the epidemiology of the so-called NDM-1 harbouring bacteria to prevent its further spread and to place effective control measures. The present study describes the use of the real-time polymerase chain reaction (PCR) assay for the detection of the bla NDM-1 gene using TaqMan probes among clinical isolates. Materials and Methods: Clinical isolates of Escherichia coli (11 strains), Klebsiella pneumoniae (17 strains) and Acinetobacter baumannii (six strains) that were resistant to either of the carbapenems (meropenem or imipenem) were included in the study. The presence of carbapenemases in such strains was confirmed using the modified Hodge test. A real-time PCR assay was optimized for the detection of NDM-1 using a cloned synthetic gene fragment followed by testing of the clinical isolates. The findings were further confirmed using PCR and gene sequencing. Results: TaqMan probe assay displayed a good detection limit with analytical sensitivity of the assay up to 10 copies of bla NDM-1 gene per reaction. The isolates of E. coli and K. pneumoniae revealed narrow range crossing point values (Cp values) between (12-17) cycles (mean Cp value 14), indicating number of bla NDM-1 gene copies of 106-108. The wider range of Cp values (15-34) cycles with a higher mean Cp value (23.6) was observed in A. baumannii with number of bla NDM-1 gene copies of 103-108. Conclusions: The study demonstrates that real-time PCR assay based on TaqMan chemistry is a useful technique for the detection of bla NDM-1 harbouring clinical isolates of E. coli, K. pneumoniae and A. baumannii. The assay has great precision in measuring the number of bla NDM-1 gene copies per specimen of DNA.

The effect of fissure morphology and eruption time on penetration and adaptation of pit and fissure sealants: An SEM study.

This study was designed to examine the effect of fissure morphology on penetration and adaptation of fissure sealants and their relationship with the eruption time of tooth. Materials and Methods: One hundred and fifty extracted molars and premolars were divided into two groups on the basis of their eruption time. The two groups were further divided into five subgroups on the basis of fissure morphology. An scanning electron microscopic analysis of penetration and adaptation of sealant was done. Observations and Results: V- and U-shaped fissures were found to have the maximum penetration. Penetration was very poor for I- and IK-types of fissures. No significant difference in penetration was found in relation to eruption time. Adaptation of sealant was not affected by any of the factors. Conclusion: Even the well-applied sealant does not necessarily provide complete obturation of pits and fissures, thus necessitating periodical clinical observation to determine the success or potential failure of the sealant treatment.

Mycotic infections of brain.

Six cases of mycotic infectation of the brain are presented. All the patients were in the age group ranging from 18 years to 38 years. the duration of clinical symptoms varied from 6 days to 7 months. Computerized tomographic visualization of brain revealed a mass lesion in all. Operative findings were suggestive of tumour in 3 cases. All the patients were non-immunocompromised. There was history of previous ear infection and sinusitis in one case. Histopathological examination of biopsy tissue showed dichotomously branching septate fungal hyphae highlighted by special stains like methanamin silver and PAS in all cases.

Mixed germ cell tumour of the pineal region: a case report.

An intracranial mixed germ cell tumour with germinoma and teratoma components is reported. The patient presented with parinaud's syndrome and precocious puberty. The treatment involved partial surgical debulking followed by whole brain radiotherapy (4500 cGY in 25 fraction over 5 weeks) and chemotherapy (consisting of cisplatin and etoposide). Post treatment MRI showed no residual lesion. The controversies in the management are discussed.

Acute pneumonitis with pulmonary hemorrhage an uncommon and potentially fatal complication of systemic lupus erythematosus: a case report.

Acute pneumonitis with diffuse alveolar haemorrhage is potentially fatal. When it occurs in a patient of systemic lupus erythematosus, the primary disease itself may be responsible for it; rather than any complicating infection or metabolic/physiological derangement. Diagnosis of primary pulmonary involvement by systemic lupus erythematosus can only be made on open lung biopsy coupled with immunofluorescent and/or ultrastructural studies. Early diagnosis of acute pulmonary complications in systemic lupus erythematosus patients is essential as specific management is reported to improve the chances of recovery.

Synchronous adenocarcinoma and MALT lymphoma of stomach.

We describe a patient in whom adenocarcinoma and lymphoma occurred simultaneously in the stomach. She presented with pain and lump in the epigastrium with history of hematemesis. Endoscopy revealed a growth involving the lesser curvature, and biopsy showed poorly differentiated carcinoma. Histological examination of the gastrectomy specimen showed synchronous diffuse adenocarcinoma with primary lymphoma of MALT type. The latter entity is known to be associated with Helicobacter pylori infection.

Epithelioid granulomas in Hodgkin's disease--prognostic significance.

Prognostic significance of non-caseating epithelioid granulomas in association with Hodgkin's disease has been studied. Such granulomas were found in 15 of the total of 104 cases of Hodgkin's disease encountered between Jan. 1981 and June 1990. These 15 patients were compared with 30 concurrent patients of Hodgkin's disease who did not have associated granulomas. All the patients were initially staged, treated and followed up for a period of two years. There was no significant difference in overall survival rate between the granuloma group and the control group. However, in relapse free survival rate in advanced stages of the disease (III & IV), although the difference between granuloma group and the control group was not statistically significant (p = 0.11), yet the relapse free survival curves revealed a tendency towards better survival with lesser number of relapses and longer remissions in granuloma group.

Bone marrow involvement in Hodgkin's disease -- clinicopathological study of seven cases.

Bone marrow involvement was observed in 7 cases amongst a total of 75 cases of Hodgkin's disease encountered between January 1981 and June 1990; thus representing an incidence of 9.3 percent. Analysis of these 7 cases showed male preponderance. The age ranged from 26 to 74 years. All patients presented within seven months of onset of symptoms; and all had symptoms at presentation. Lymphadenopathy was found in all the cases, splenomegaly was present in six; and hepatomegaly in 4 cases. Mixed cellularity and lymphocytic depletion subtypes showed the highest frequency of marrow involvement (6 cases; 85.7 percent). Alkaline phosphatase was raised in 6 cases (85.7%). All cases received standard combination chemotherapy with or without local radiation therapy. Two years follow-up revealed an overall survival of 42.9%. Replace free survival at the end of two years was zero, as all the cases relapsed within seven months of initiation of treatment.

Hodgkin's disease: a clinicopathologic study.

One hundred and four cases of Hodgkin's disease diagnosed between July 1981 and June 1991 have been analysed. There was a definite male preponderance. Majority of the patients (82.7%) were below the age of 50 years. Mixed cellularity was the most common type (57.7%). It was followed by both nodular sclerosis and lymphocyte predominant types (16.3% each). Lymphocyte depletion Hodgkin's disease, the most aggressive variant, was the least common (9.7%). The detailed observations, as compared to the previous studies in this region as well as in other parts of the world have been presented and discussed.

Effect of aerosolised verapamil and nifedipine in bronchial asthma.

Twenty patients of bronchial asthma were given 5 mg of either nifedipine or verapamil as aerosol therapy in a randomized fashion. Both drugs showed a statistically significant improvement in FVC, PEFR at zero hours lasting through 90-120 minutes. Changes in respiratory rate, pulse rate, blood pressure were insignificant with verapamil while appreciable tachycardia and transient fall in BP occurred with nifedipine therapy.

Hepatic involvement in tuberculosis.

Seventy one cases of pulmonary and extrapulmonary tuberculosis (confirmed by clinical, sputum examination, Skiagram, lymph node or other organ biopsy) and 13 cases of PUO suspected to be tubercular in origin were included in the study. Besides, thorough clinical workup, LFTs and liver biopsy were also done in all cases. LFT alterations were seen in only 5 (7%) cases of tuberculosis while in PUO group, 3 (23%) cases showed deranged LFT. In all, alteration in SGOT/SGPT was most frequent observation. In Liver biopsy, histopathological changes were seen in 63 percent (45 out of 71) of cases of tuberculosis and in 46.16 percent cases of PUO. In tubercular cases, various abnormalities observed were caseating granulomas: 18.30 percent, nonspecific inflammatory infiltration: 25.32 percent, focal Kupffer cell hyperplasia: 11.2 percent and fatty changes: 8.45 percent. Four of the PUO cases, showing Kupffer cell hyperplasia and non-specific inflammatory infiltration responded to antituberculous therapy. Thus it is surmised that though the caseating granuloma is the hallmark of tuberculosis, but nonspecific inflammatory hepatitis and retothelial tubercle nodules may be the forerunner in the evolution of granuloma.

Congenital hemihypertrophy: report of four cases.

Congenital hemihypertrophy is a rare condition. We came across four such cases within a period of six months. These cases are presented here and their unusual features highlighted.

In vitro sensitivity of antifungal agents.

In vitro sensitivity tests indicated econazole to be the best drug against all 30 isolates of Trichophyton species followed by miconazole, clotrimazole and tolnaftate. Econazole inhibited the growth of all isolates of Trichophyton species in low concentrations of 2-4 micrograms/ml except one isolate of T. rubrum which was inhibited by a concentration of 8 micrograms/ml. Miconazole was also able to inhibit growth of 73.3 per cent isolates of Trichophyton species in low concentrations of 2-4 micrograms/ml. On the other hand, only 13.3 per cent isolates were found sensitive at 2-4 micrograms/ml concentration to tolnaftate and clotrimazole. Relative resistance was encountered by 7 isolates (23.3%) to tolnaftate and 2 isolates (6.6%) to clotrimazole. Isolates of T. tonsurans were resistant to tolnaftate, whereas, the isolate of T. mentagrophytes was sensitive to all the drugs in low concentrations.