Immunohaematological conundrums in obstetrics from transfusion services viewpoint in Armed Forces setup.

Background: Obstetrics as a speciality has a very long association with the transfusion services and poses its own set of immunohematological (IHL) challenges. A study was carried out to evaluate the spectrum of IHL issues in obstetrics in our setup and to suggest a way forward. Methods: This study was carried out in a transfusion services setup catering to antenatal care (ANC) clientele in two tertiary-level health care setups. Samples were collected from all ANC patients requiring transfusion and patients reporting for Indirect Coombs Test (ICT). Data included ICT positive cases with implicated alloantibodies, those requiring specialised procedures and the foetal outcome. The results were described using descriptive statistics by frequencies and percentages. Results: A total of 4683 eligible samples were included in the study, out of 21,893 antenatal patients visiting our setup during study period. One hundred thirty-six ANC patient samples were found to be ICT positive. The most common single alloantibody was anti-D (n = 77, 57.5%). Double antibody positivity was found in 28 patients. Multiple alloantibodies were found in 1 patient. Up to 48% of these allo-anti D cases necessitated specialised procedures. Conclusion: The IHL issues of obstetrics faced in our setup are no less than that in Indian population. We have much higher frequency of double alloantibody in our ANC population. The authors propose that all multiparous ANC patients, especially with a history of transfusion, irrespective of Rh D status should be screened for irregular alloantibodies to circumvent these issues and the last-minute rush for provision of compatible units.

A pilot study on DNA hypermethylation status in promoter region of P16 gene in patients with sporadic breast cancer.

Background: Epigenetic modification of cancer-related genes plays a role over and above their genetic alterations and contributes to the tumor initiation and progression of breast cancer. Promoter methylation of tumor suppressor genes is one such epigenetic modification, which can be potential biomarker. In this study, promoter methylation status of p16 gene was studied in blood samples of patients with breast carcinoma. Methods: Seventy-five patients, freshly diagnosed with carcinoma of breast and 20 age and sex matched healthy control subjects were recruited for the study. DNA extracted from EDTA blood sample was bisulfite converted and subjected to methylation-specific PCR to amplify the p16 promoter region. Results: Out of 75 patients, 25 (33%) patients showed hypermethylation in promoter region of p16 gene, which was statistically significant in comparison with the control group (p < 0.05). In subgroup analysis, lymph node involvement, cancer grade, and histopathological finding did not show any difference with methylation status of p16 promoter. Conclusion: Significant hypermethylation of p16 promoter region in the blood of histopathologically proven cases of breast cancer was observed suggesting promoter hypermethylation of p16 may be a possible mechanism accounting for sporadic carcinoma of breast.

Major and minor blood group phenotyping and database generation for recruits : A pilot study.

BACKGROUND: Alloimmunization to minor blood group antigens is a problem that has for long befuddled the blood bankers, requiring blood group phenotyping of target populations worldwide. However, the same exercise had been lacking in our Armed Forces population necessitating this pilot study. METHODS: A total of 2000 recruits of a regimental center in northern India were phenotyped for major and minor blood group antigens including ABO, Rhesus (D, C, c, E, and e antigens), Kell (K), MNSs (S and s antigens), Kidd (Jka and Jkb), and Duffy (Fya and Fyb) using commercially prepared polyclonal antisera on a fully automated system based on electromagnetic technology. Typing for M and N antigens (MNSs system) was performed using the tube method using company provided specific polyclonal antisera. The results were also compared with that of Indian and other populations of the world. RESULTS: Blood group antigen frequencies observed in our study population were mostly in sync with the prevalence rates of major and minor blood group antigens reported in other studies; however, in certain cases, they were also at variance. In our study, Rh D-negative antigen, JKa-Jkb- phenotype frequency were comparatively much higher than those reported in other Indian studies. CONCLUSION: To conclude, ours was a pilot study to establish the database of major and minor antigens of Armed Forces combatants. However, it is recommended that if the same effort can be replicated at the other transfusion centers and major hospitals of Armed Forces, it will vastly benefit their alloimmunized patient clientele needing lifesaving transfusions.

Double-positive in triple-negative? How significant is basal cytokeratin expression in breast cancer?

BACKGROUND: Basal-like breast cancer has an unfavorable prognosis. Immunohistochemically, they are predominantly estrogen receptor (ER), progesterone receptor (PR) and CerbB2 receptor (HER2)-negative, show expression of Cytokeratins (CKs) 5/6, CK14, CK 17 and P-cadherin and are associated with germline BRCA1 mutations. Immunohistochemistry (IHC) is an easily available and relatively inexpensive technique that can detect this cancer subtype, and patients can benefit from aggressive management protocols. The aim of this study was to evaluate the expression of CK 5/6 and CK14 in breast cancer and its correlation with age, tumor grade, tumor size, histomorphological pattern, nodal status, ER, PR, HER2/neu, and Ki-67 index. METHODS: Fifty treatment-naїve patients of breast carcinoma who underwent surgery constituted the study group. No core cut biopsy specimens were considered. Histopathological examination along with IHC was performed for CK5/6, CK14, ER, PR, HER2/neu, and Ki-67. Comparison between the expression of CK5/6 and CK14 with age, tumor size, tumor grade, histological subtype, nodal status, ER, PR, HER2/neu, and Ki-67 expression was performed using SPSS 20 version software. RESULTS: Twenty-six percent of cases showed expression of CK5/6 and CK14. CK5/6 and CK14 expression correlated strongly with ER/PR negativity, young age, and Ki-67 proliferative index greater than 15%. No significant association with HER2/neu negativity was demonstrated. Contrasting results were obtained between CK5/6 and CK14 expression with respect to tumor grade and lymph node status. CONCLUSION: IHC can be used to identify patients with basal phenotype breast cancer with good sensitivity and specificity, and such patients can benefit from aggressive management.

Rapid in-situ hybridization and immunohistochemistry: a pilot comparative study of two rapid diagnostic techniques for establishing monoclonality in plasma cell dyscrasias.

BACKGROUND: Light chain restriction needs to be established on the paraffin embedded tissue in certain types of plasma cell dyscrasias when serum levels of monoclonal immunoglobulins and light chain assays in the urine and serum may be normal. Rapid-in-situ-hybridisation (RISH) is thought to be a superior to immunohistochemistry (IHC) for kappa and lambda staining due to brighter and crisp staining without any background. METHODS: Fifty cases were included in this pilot study. Serum light chain restriction status of the case was taken as gold standard. The results of standard IHC for kappa and lambda immunoglobulins on the bone marrow biopsy of these cases was compared with RISH performed by the two commercially available kits. The results of the two methods were compared for sensitivity, need to repeat the test and background staining. RESULTS: The study found that in IHC first run sensitivity was 58% which improved to 88% after the second run. For RISH the sensitivity was 100%. CONCLUSION: Rapid-in-situ-hybridisation (RISH) is a superior technique to IHC for detecting kappa and lambda light chain in plasma cells. The test is as labour intensive and time consuming as the routine IHC but has no background staining with more bright and crisp staining quality.

A study of BRAF mutation in colorectal carcinoma in Indian population.

CONTEXT: BRAF mutation has been extensively studied and associated with various tumors. Targeted therapeutic intervention against BRAF mutation is established modality against many such tumors. Various studies have estimated that the prevalence of BRAF mutation in colorectal carcinoma (CRC) is 5%-25%. Considering epidemiology differences from Western population and paucity of studies on BRAF mutation in CRC in Asian patients, the present study was done to study the BRAF mutation in CRC in Indian population. AIMS: The aim is to study the distribution of BRAF mutation and its correlation with the American Joint Committee on Cancer (AJCC) stage, grade, and other clinicopathological parameters in CRC. SETTINGS AND DESIGN: This was a cross-sectional study. SUBJECTS AND METHODS: Immunohistochemistry study was done using BRAFV600E monoclonal antibody (Clone VE1) for 65 consecutive cases of CRC in a tertiary care center. The results were statistically analyzed using SPSS version 2.0. RESULTS: This study found that BRAF mutation is not significantly present in CRC as only 4.6% of cases were positive for BRAFV600E mutation. However, there was statistically significant relation between increasing AJCC stage and BRAF mutation. CONCLUSIONS: This study concluded that BRAF mutation is not prevalent in Indian population with CRC. However, it is significantly related with advanced AJCC stages.

A study of therapy targeted EGFR/ALK mutations in Indian patients with lung adenocarcinoma: A clinical and epidemiological study.

BACKGROUND: Established predictive biomarkers for Non-Small Cell Lung Carcinoma (NSCLC) include sensitizing Epidermal Growth Factor Receptor (EGFR) mutations and Anaplastic Lymphoma Kinase (ALK) fusion oncogene. The primary aim of the study is to ascertain the prevalence of EGFR mutation and ALK gene rearrangement in patients of lung adenocarcinoma in Indian population and the second objective is to impress upon the importance of adequate processing of limited tissue samples. METHODS: Histopathologically confirmed cases of lung adenocarcinoma, whose tumour had been tested for both EGFR and ALK gene mutations, were included in this study. The EGFR mutations were analyzed using PCR and Gene Sequencing. ALK fusion oncogene was found by Fluorescence In Situ Hybridization (FISH) technique using kit of Vysis LSI ALK Dual colour Break Apart Rearrangement probe. RESULTS: A total of 152 cases of lung adenocarcinoma were included. Out of which, 92 (60.5%) were male and 60 (39.5%) were female. After exclusion of 17 cases due to unsatisfactory result, EGFR mutations were found positive in 35.5% cases (48/135). ALK gene rearrangement was found in 7.6% (10/131) after excluding 21 cases with unsatisfactory result. CONCLUSION: EGFR mutations and ALK gene rearrangement was found to be mutually exclusive. Incidence of EGFR mutations (35.5%) is much higher in Indian population than in Caucasians (13%) and is close to the incidence in East Asian countries. The 7.6% incidence of ALK fusion oncogene in Indian patients establishes the importance of molecular studies to give maximum benefit of targeted therapy to the patients.

Adult-onset Still's disease with myocarditis and hemophagocytic lymphohistiocytosis: Rare manifestation with fatal outcome.

Adult-onset Still's disease (AOSD) is a rare inflammatory disorder of unknown etiology characterized by fever, evanescent pink salmon rash, arthritis, and multiorgan involvement. Here, we report an unusual manifestation of AOSD in a 40-year-old male who presented to our hospital with pyrexia of unknown origin and rash of 3 weeks duration. All his serological investigations and imaging studies were unremarkable. He was fulfilling clinical and laboratory criteria as per Yamaguchi for AOSD and was managed for the same. Our patient did not respond well to the treatment, had a downhill course, and succumbed to his illness. Autopsy confirmed myocarditis and florid bone marrow reactive hemophagocytosis as the cause of his death.