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.

Clinicopathological and immunohistochemical analysis of Sarcomatoid carcinoma of head and neck mucosal region: a retrospective analysis

Background: Sarcomatoid carcinoma is a biphasic tumour comprising both of malignant epithelial and mesenchymal elements derived monoclonally from same stem cells. These are unusual variants of squamous cell carcinoma and constitute less than 1% of the head and neck mucosal tumors. Only few studies have been published and needs more understanding to establish treatment guidelines. The aim of this study was to review the cases of carcinosarcoma arising from mucosal sites of head and neck and study their clinical, histological and Immunohistochemical features.Methods: Retrospective data and slides of histologically proven sarcomatoid carcinoma over a period of thirty -four months between January 2016 - October 2018 were retrieved and evaluated for various clinical and histopathological parameters.Results: Total of 22 cases were included in the study and the mean age of presentation was 58years with male: female ration 2:1. Most of the patients (81.8%) presented with a mass lesion of less than 6 months duration. The most common site was oral cavity (68.1%) followed by larynx (22.7%). Clinical stage was known in eleven cases. One case presented with pulmonary metastasis. Histopathologically, epithelial differentiation was identified in nine cases (41%) on morphology and in thirteen cases could be highlighted by cytokeratin positivity. The Mesenchymal component was arranged in sheets (63.7%) and fascicles (31.8%). Marked anaplasia and brisk mitosis wereseen in 54.5% and 19.3% respectively.  On immunohistochemistry all 22 cases were positive for Vimentin, twenty cases were positive for cytokeratin/EMA and aberrant mesenchymal markers were expressed in 10% of cases.  Follow up was available in eighteen cases out of which fourteen cases died within one year of diagnosis.Conclusions: Diagnosis of sarcomatoid carcinoma is challenging especially on small biopsy because of overlapping features with other spindle cell tumors. Understanding the clinicopathological features facilitates their diagnosis and effective clinical management.

Process optimisation, characterisation and evaluation of resveratrol-phospholipid complexes using Box-Behnken statistical design.

With the advent of 21st century, researchers worldwide have extensively reviewed herbs and botanicals for their marked clinical efficacy. It has been estimated that most of the newly discovered compounds offer poor bioavailability due to their low aqueous solubility. Phospholipid complexation of the drug often helps to improve its water solubility and enhance the bioavailability. This study includes optimization of resveratrol-phospholipid complexes using a 3-factor, 3-level box-behnken design (15 batches). Three independent variables i.e. phospholipid-resveratrol ratio, refluxing temperature and reflux time were optimized for two dependent variables, i.e. yield and entrapment efficiency (EE). Complexes were prepared by refluxing stoichiometric ratio of Phospholipon 90G and resveratrol in dichloromethane and retrieved by precipitation with n-hexane. Complexation was confirmed by Fourier Transform Infra-Red (FTIR) spectroscopy. The data was suitably used to explore quadratic response surfaces and construct second order polynomial models with Design Expert®. Formulation with highest desirability (D=0.994) was selected as optimum and prepared using 1.5:1 Phospholipon 90G-resveratrol ratio (X1) at 59.4°C temperature (X2) and 4 h time (X3) to give maximum yield and entrapment efficiency. Analysis of variance (ANOVA) was also found to be significant for both the responses. Complexes were optimised for good yield and EE. The partition coefficient was lowered to 2.25 hypothesizing good passive absorption.