BRCA1 promoter methylation & its immunohistochemical correlation in sporadic breast cancer

Background & objectives: Studies have shown that apart from hereditary breast carcinomas, breast cancer susceptibility gene 1 (BRCA1) mutations conferring to its loss are seen in sporadic breast carcinomas (SBC) as well. The aim of the present study was to assess BRCA1 methylation in females presenting at King George’s Medical University, Lucknow, with SBC by both immunohistochemistry (IHC) and methylation PCR with respect to hormonal profile and various morphological prognostic parameters. The primary objective was to look for the association between BRCA1 protein expression and DNA promoter methylation. Methods: 81 mastectomy specimens from SBC of invasive breast carcinoma (no special type) were included in this study. After a detailed morphological assessment, formalin fixed paraffin embedded tissue from a representative tumour area was selected for BRCA1 IHC by heat-mediated antigen retrieval under high pH and DNA extraction and further bisulphate treatment. BRCA1 was studied for methylation by methylated and unmethylated PCR-specific primers. Results: BRCA1 promoter methylation was present in 42/81 (51.9%) participants, with significant BRCA1 protein loss (72.7%; P=0.002). A significant association between BRCA1 loss and hormonal profile was found (P=0.001); maximum in triple negative breast carcinoma (TNBC) (72%; 18/25). Most of the TNBC also harboured methylation (68%). Although not significant grade II and III tumours, lymph vascular invasion, ductal carcinoma in situ, and nodal metastasis (?3) were seen in a higher percentage in methylated tumours. Mortality in SBC was significantly associated with BRCA1 loss (30.3%; P=0.024). Interpretation & conclusions: Study results highlight the concept of “BRCAness” in SBC as well. Hence, we can confer that identification of BRCA1 loss in SBC can make it a perfect candidate for poly ADP- ribose polymerase inhibitors or cisplatin-based therapy like hereditary ones.

Disparity between cost variation of antibiotics in northern and western India: A cross - sectional study

Background - About 58% of India’s health expenditure is out of pocket expenditure. There is a wide variation in the cost of different brands of the same generic drug. Prescription of expensive brands of antibiotics contributes to the development of antibiotic resistanc e. There is literature available on the cost variation and cost ratio of antibiotics but none on the regional disparity. Materials and Methods - An observational cross - sectional study design was adopted. The study was conducted at the medical stores of Ali garh (Uttar Pradesh) and Mumbai (Maharashtra). The maximum and minimum cost/unit in Rupees (INR) of each antibiotic manufactured by multiple pharmaceutical companies was noted. Furthermore, Cost per daily defined dose (cost/DDD) was calculated for the most expensive and least expensive drug. Results - ALIGARH - The highest percent cost variation of 1897.14 and cost ratio of 19.97 was found for Piperacillin 4000mg + Tazobactam 500mg tablets. The least percent cost variation of 48.92 and cost ratio of 1.49 was found for ciprofloxacin 200mg vial. MUMBAI - The highest cost variation of 3031.05 and cost ratio of 31.31 was found for cefixime 200 mg tablets. The least percent cost variation of 6.43 and cost ratio of 1.06 was found for ciprofloxacin 200 mg vial. Concl usions - There is disparity in the cost variation and ratio between the two cities Aligarh and Mumbai in this study, which can be attributed to availability of different brands, under a generic group, in the different regions of the country

Chronic suppurative otitis media: a clinico-microbiological menace.

Background: Chronic Suppurative Otitis Media (CSOM) is an important cause of preventable hearing loss. Global emergence of resistant strains is of great concern. The aim of the present study was to determine the etiology and antibiotic sensitivity pattern of bacterial isolates from CSOM cases with special emphasis on ESBL (Extended Spectrum Beta- Lactamases) and AmpC beta lactamases. Methods: Patients with sign and symptoms suggestive of CSOM, ESBL (Extended Spectrum Beta-Lactamases), AmpC beta lactamases and MBLs (Metallo beta lactamases) were included. Two ear swabs were taken from all the patients and cultured on blood agar and MacConkey agar. Bacterial identification of isolates was done using standard biochemicals. Antimicrobial susceptibility was performed by Kirby-Bauer's disc diffusion method as per the Clinical Laboratory Standards Institute (CLSI) guidelines using antibiotic discs (HI MEDIA). Results: Out of 130 patients, 110(84.62%) had bacterial growth. The common pathogenic species were Pseudomonas aeruginosa 36(37.89%), Staphylococcus aureus 31(32.63%), Citrobacter koseri 9(9.47%) and Proteus vulgaris 6(6.32%). P. aeruginosa showed maximum sensitivity to colistin (94.4%), polymixin-B (91.3%) and imipenem (91.3%). Gram positive cocci showed maximum sensitivity to vancomycin (99%). MRSA (Methicillin Resistant Staphylococcus aureus) and HLAR (High Level Aminoglycoside Resistance) were detected in 9(29%) S. aureus and 1(50%) Enterococcus faecalis respectively. ESBL and AmpC were detected in 11(18.3%) and 12(20%) Gram negative bacteria, respectively and MBL producer was not detected. Conclusion: P. aeruginosa was found to be the most common isolate in CSOM cases and colistin, polymixin-B and imipenem was found to be most effective antibiotics.

Prevalence of genotype D in chronic liver disease patients with occult HBV infection in northern region of India.

Background: Etiology of nearly 30% cases of chronic viral hepatitis remains undetected. Occult HBV infection (OBI) has emerged as an important clinical entity in this scenario. Apart from prevalence and clinical outcome of OBI patients genotype was determined in northern region of India. Materials and Methods: A total of 847 patients with chronic liver disease (CLD) were screened for common viral etiologies and others serological markers of HBV. Amplifi cation of surface, precore and polymerase genes of HBV was performed in patients negative for other etiologies. Genotyping and sequencing of the precore region was performed for OBI cases. Results: Twenty-nine (7.61%) cases of OBI were identifi edof which 9 had chronic liver disease (CHD), 11 liver cirrhosis (LC) and 9 hepatocellular carcinoma (HCC). Majority of OBI cases were detected by amplifi cation of surface gene 26 (89.6%), followed by pre-core gene 12 (41.3%). Their liver functions tests were signifi cantly deranged in comparison to overt HBV cases. IgG anti HBc was present in 8 (27.6%) OBI cases. Mutation was observed in 8 (32%) in pre-core region at nt. 1896 of overt HBV cases. Genotype D was the predominant genotype. In conclusion: OBI in our study was characterized by predominance of genotype D and more severe clinical and biochemical profi le in comparison to overt HBV. IgG anti HBc positivity could be utilized as a marker of OBI. We recommend use of sensitive nested PCR for diagnosis of OBI, amplifying at least surface and precore gene.