Anti-Histone H3.3 G34W antibody is a sensitive and highly specific immunohistochemistry marker for the diagnosis of Giant cell tumor of bone. A validation based on analysis of 198 cases from a single centre in India

Context: The diagnosis of giant cell tumor of bone (GCTB) is difficult in small biopsies with unusual age of presentation, location, and extensive secondary changes. Most of the GCTBs harbor H3F3A G34W mutations with a subset of cases showing alternate G34V, G34R, and G34L mutations. Objectives: To analyze the expression of anti-histone H3.3G34W antibody in different cellular components of GCTB across different locations and presentations (including the unusual ones) and validate the utility of this antibody in the diagnosis of GCTB and differentiate it from the other osteoclast-like giant-cell-rich lesions. Design: Immunohistochemistry was performed using anti-histone H3.3G34W antibody in the diagnosed cases of GCTB (136 cases of GCTB from 133 patients, including two malignant GCTBs) and other giant cell-containing lesions (62 cases). The presence of unequivocal crisp nuclear staining was considered positive. Results: Immunohistochemistry revealed unequivocal nuclear positivity in the mononuclear cells in 87.3% of the cases of GCTB. Of these, most showed diffuse expression with moderate to strong intensity staining. The positive staining was restricted to the nuclei of mononuclear cells with the nuclei of osteoclastic giant cells being distinctly negative. In addition to conventional GCTBs, two cases each of multicentric and malignant GCTB showed positive staining. The other giant-cell containing lesions were distinctly negative. The present study showed a sensitivity of 87.3% with specificity and positive predictive value of 100%. Conclusion: The anti-histone G34W antibody is a highly sensitive and specific marker for the diagnosis of GCTB and differentiating it from its mimics. The positive staining is restricted to the mononuclear cell component of GCTB with sparing the osteoclastic giant cells further reiterating the fact that the mononuclear stromal cells are the true neoplastic component of GCTB.

Assessing The Rationale Of Fdc Containing Ofloxacin And Azoles: Dissolution, Permeation And Antimicrobial Studies

Objective: To study fixed-dose combinations (FDC) of antibacterial and antiprotozoal products (ofloxacin and azoles), prescribed for the treatment of diarrhea. Methods: Rationality of these FDC products was verified by assessing parameters such as drug content and release by assay and dissolution tests, respectively mentioned in the Indian Pharmacopoeia (IP). Amount of drug solubilized and permeated as per the Biopharmaceutics Classification System (BCS) was determined. Ex vivo permeation study was performed on the gut of goat using the everted gut sac technique. Antimicrobial efficacy in terms of minimum inhibitory concentration (MIC) was assessed using agar well diffusion method against Shigella boydii, the causative agent for diarrhea. Comparative studies were performed on an individual as well as combination doses of antibacterial and antiprotozoal products for the synergistic effects to assess the rationale of these FDC. Results: The BCS solubility of ciprofloxacin (CPX), norfloxacin (NFX) and tinidazole (TNZ) was high in acidic medium (pH 1-5) and decreased at pH above 5. The assay studies showed that the individual drug contents of FDC were within the IP limits. In vitro dissolution results for both, individual drugs and their combination illustrated 99 % drug release within 30 min in 0.01N HCl. Ex vivo permeation of TNZ was higher than CPX and NFX in individual drugs. No significant change in the permeation rate was observed for individual drugs and their FDC. CPX and NFX exhibited more antimicrobial activity in terms of inhibitory zones than their FDC with antiprotozoal TNZ, above 2.5 µg/ml MIC. The pharmaceutical, biopharmaceutical and antimicrobial evaluation study showed the similarity of FDC with the individual drugs. Conclusion: The study showed no significant data to justify the therapeutic advantage of FDC over individual drugs.

Characterization of Xanthomonas campestris pv. viticola causing bacterial leaf spot of grapes in Maharashtra, India

Aim: The aim of the present study was to isolate, characterize and identify pathogenic bacteria from bacterial leaf spot infected grape vineyards of Maharashtra. Methodology: Collected diseased samples were subjected to isolation. All isolates were further subjected to different morphological and biochemical characterization. These isolates were also used to standardize inoculation methods to study host range and test pathogenicity to confirm infectivity. Genomic DNA was isolated from bacteria for phylogeneitc analysis using 16 rDNA. Results: The disease is characterized by water soaked, angular leaf spot which later turns to irregular, dark brown to black necrotic region on the leaf surface. Microbial studies confirmed that it is a Gram negative, rod shaped bacterium with white, mucoid, glistening and convex circular colonies on artificial medium. Pathogenicity test was performed to confirm virulence. Phylogenetic analysis of 16S rDNA gene exhibited more than 99% similarity with other Xanthomonas campestris pv. viticola. The bacterium was found resistant to vancomycin, peniciliin and oxacillin antibiotics. The bacterium is also capable of infecting Mangifera indica and Citrus limon along with eight weed species.

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.

Clinical profile of non-O1 strain-O139 of Vibrio cholerae in the region of Ambajogai, Maharashtra.

OBJECTIVES: To study clinical profile of the newly emerged novel strain non-O1, O139 of Vibrio cholerae, in the region of Ambajogai, District Beed of Maharashtra. METHODS: Out of 208 patients of acute gastroenteritis, 41 revealed to be positive for Vibrio cholerae by recommended method of stool examination. All the strains were sent to National Institute of Cholera and Infectious Diseases, Calcutta for confirmation. RESULTS: Out of 41 cases, 12 were of Vibrio cholerae O1, 29 Non-O1, of which nine found to be O139 strain. All patients were from 2-80 years of age with low-socioeconomic status and maximum incidence was in August (64.70%), presented with severe rice watery loose motions. Vomiting was observed in 26 (63.41%), more so in patients of O139 infection (88.88%) than four (33.33%) of O1 infection. Sweating was observed in three patients (33.33%) of O139 infection, cramps in gastrocnemis muscles in three patients (33.33%) of O139 infection and two (16.66%) of O1 infection. Signs of dehydration were mild to moderate in four patients (33.33%) of O1 infection; severe dehydration in six (66.66%), moderate in two (22.22%) and mild in one patient (11.11%) of O139 infection. While dehydration was severe in four (20%), moderate in one (5%) and mild in three patients (15%) of Non-O1 infection (excluding O139 cases). Clinical features were more severe in patients of serotype O139 than the patients of O1 and Non-O1 (excluding O139 cases). However all patients responded to intravenous fluids, oral rehydration and antibiotics (tetracycline) within 24-48 hours without any mortality. CONCLUSIONS: This study reflects the first emergence of Non-O1, strain O139 during the year 1997 with severe and critical clinical features in Ambajogai region causing high morbidity in the form of severe dehydration and peripheral circulatory collapse which requires early and correct diagnosis and prompt treatment.

Left ventricular M-mode echocardiographic measurements of Indian population.

The left ventricular dimensions of 506 normal healthy subjects (men = 317, women = 189) in the age group of 17 to 80 years were measured echocardiographically. The measurements were found to differ significantly from the western data. A separate limit for left ventricular echocardiographic parameter for the normal Indian population was established.

Left ventricular mass in normal Indian population.

A prospective study was conducted to ascertain the distribution of the left ventricular mass in normal adult Indian population and to establish the upper limit of the same. Four hundred and eighty normal subjects (men = 310; women = 170), in the age group of 18 to 60 years were screened echocardiographically, to estimate the left ventricular mass. The left ventricular mass in men was found to be 124 +/- 32 gm whereas in women it was 93 +/- 37 gm. The upper limit of the left ventricular mass (mean +/- 2 SD) was found to be 189 gm and 167 gm, for men and women respectively. The left ventricular mass in both men and women correlated with the body weight (r = 0.51, p less than 0.001; r = 0.22, p less than 0.001), height (r = 0.27, p less than 0.001; r = 0.22, p less than 0.01), and the body surface area (r = 0.49, p less than 0.001; r = 0.27, p less than 0.001), whereas it was found to be independent of age (r = 0.01, p = NS; r = 0.10, p = NS). In men alone the diastolic blood pressure influenced the left ventricular mass (r = 0.27, p less than 0.001), otherwise the left ventricular mass was independent of systemic blood pressure variations within the normal range. The left ventricular mass in the present study differs significantly from the Western population.