Correlation NKX2.2 IHC and EWSR1 break-apart FISH in the diagnosis of Ewing sarcoma: Can combined NKX2.2 and CD99 immunoexpression obviate or minimize the need of FISH testing? First assessment study from Indian tertiary cancer care center

Context: Ewing sarcoma (ES) are malignant small round cell tumors (MSRCT) characterized by rearrangements of EWSR1 gene. Although gold standard for diagnosis is detection of specific fusion genes by molecular testing, these ancillary tests are costly and only available in limited number of settings. There is a persuasive evidence for reliability of NKX2.2 immunohistochemistry (IHC) as a surrogate marker for EWSR1 gene rearrangement in ES. Aims: The aim of this study is to correlate the NKX2.2 immuno-expression with genetically confirmed ES cases and also to assess the reliability and accuracy of NKX2.2 along with combined positivity of NXX2.2 and CD99 in diagnosing ES and differentiating it from other relevant histological mimics. Settings and Design: The present study is a retrospective study conducted over a period of 6-year duration in a tertiary cancer care center. Methods and Material: We evaluated NKX2.2 immunoexpression in 35 genetically confirmed cases of ES and also in pertaining differential entities (n = 58) of ES including rhabdomyosarcoma (n = 20), lymphoblastic lymphoma (n = 14), Wilms tumor (n = 10), poorly differentiated synovial sarcoma (n = 4), small-cell osteosarcoma (n = 4), neuroblastoma (n = 5), and mesenchymal chondrosarcoma (n = 1). CD99 was performed in the category of MSRCTs showing NKX2.2 positivity to evaluate combined specificity for the diagnosis of ES. Results: Of the 35 genetically confirmed cases of ES, 29 cases (83%) showed NKX2.2-positive expression (83% sensitivity). Compared to ES, NKX2.2 was positive in only 05% cases (3/58 cases) of non-ES MSRCT. Only two of five cases of neuroblastomas and one case of mesenchymal chondrosarcoma showed NKX2.2 positivity. CD99 positivity was seen in 100% of ES and in the single case of mesenchymal chondrosarcoma. All five cases (100%) of neuroblastoma were negative for CD99. Conclusions: The presented study, which is the first from an Indian oncology center, showed NKX2.2 IHC is quite reliable in diagnosis of ES in the right clinicopathological context. With remarkable sensitivity and specificity of NKX2.2 IHC for diagnosis of ES, we propose that combined positivity of CD99 and NKX2.2 IHC can obviate or minimize the need of EWSR1 gene rearrangement molecular testing for diagnosis of ES.

Diagnostic ability of real-time quantitative polymerase chain reaction versus immunohistochemistry for Ki-67 assessment in breast cancer: An Indian perspective

Background & objectives: Breast cancer is the most common cancer of women. Inferior prognosis in some patients has been attributed to the higher proliferative capability of the tumour. Immunohistochemistry (IHC) for Ki-67, despite being a simple and cost-effective method, has not become a valid tool to evaluate this biomarker. This is ascribed to variation in pre-analytical and analytical techniques, variable expression, hotspot distribution and inter-and intra-observer inconsistency. This study was aimed at defining the analytical and clinical validity of real-time quantitative polymerase chain reaction (RT-qPCR) as an alternative to IHC evaluation. Methods: This study included a total of 109 patients with invasive breast cancers. Ki-67 IHC visual assessment was compared with the mRNA value determined by RT-qPCR. Concordance between both the methods was assessed. Receiver operating characteristic (ROC) curve analysis and Cohen's kappa value with intraclass correlation were performed. Results: The threshold value for Ki-67 by RT-qPCR obtained by ROC curve was 22.23 per cent, which was used to divide breast cancer cases into high proliferative and low proliferative groups. A significant correlation was observed between both the breast cancer groups formed using RT-qPCR threshold as well as median laboratory value of Ki-67 labelling index by IHC. Interpretation & conclusions: The study results showed a significant correlation between the two methods. While IHC is subject to technical and interpretative variability, RT-qPCR may offer a more objective alternative.

Immunophenotyping of male breast cancer - Experience at a tertiary care centre

Background: Male breast cancers (MBCs) are uncommon and account for 1% of all breast cancers. Medical conditions that increase the estrogen to testosterone ratio are implicated as the risk factors. Morphologically similar, but MBCs have biological differences compared with female breast cancer (FBC). Purpose: The present study was aimed to examine the immunophenotype of MBC, subsequent molecular subtypes, their association with clinicopathological features, and prognosis. Materials and Methods: We analyzed clinicopathological features of 42 cases of MBC, and classified them according to molecular classification using immunohistochemistry (IHC). This is the second largest study from India. Results and Conclusion: Median age of patients was 61 years (age range: 41-87 years). Invasive duct carcinoma comprised 95.2% of cases. Tumor grade II and III was seen in 50% and 47.6% of cases, respectively, and advanced stage disease (III/IV) was seen in 45.2% cases (n = 39). Estrogen receptor (ER) was positive in 97.6% cases, progesterone receptor (PR) in 83.3%, androgen receptor (AR) in 76.2%, HER2 in 4.8%, Cyclin-D1 in 92.9%, Bcl2 in 66.7%, GCDFP-15 in 23.8%, p53 in 16.7%, and Ki67 index was low (<14%) in 66.7% cases. Molecular subtyping of these cases revealed 64.3% of luminal A, 35.7% of luminal B, and no HER2 rich/driven category or triple negative case. There was no statistical significance between luminal A and B category pertaining to overall stage of tumor (P = 0.905). Lymph node metastasis was more commonly associated with luminal B category (P = 0.089). p53 positivity showed significant association with luminal A cases (P = 0.002) and nodal metastasis (P = 0.042). GCDFP-15 positivity showed significant association with higher tumor grade (P = 0.042) and stage (P = 0.047). Stage was the most significant prognostic marker (P < 0.0001). On follow-up (n = 27), all the six cases that showed recurrence/persistent disease were high stage (III/IV) on presentation.

Study of clinical profile of organophosphorus poisoning with special reference to electrocardiographic changes and electrolyte derangement

Background: Organophosphorus Compounds (OPC) are main cause of accidental and suicidal poisoning in agrarian countries like India. Aim was to study the clinical profile of OPC-Poisoning and correlate it with the electrocardiographic (ECG) changes and electrolyte abnormalities.Methods: Hundred consecutive cases admitted to Medicine Department underwent clinical examination, ECG, electrolytes, Acetyl Choline Esterase (AChE) estimation from time to time and Paradeniya Organophosphorus Poisoning (POP) score at the time of admission. All these parameter with duration of hospital stay and outcome were statistically analysed using X2 test, Fisher exact test, and inference was drawn.Results: In hundred OPC-Poisoning patients [Male (n=48), Female (n=52), M: F ratio 0.92:1] with mean age of 37.78�.95 years, commonest poison was cholropyrifos+cypermethrin and was mostly suicidal (96%). Common symptoms were sweating (48%), salivation, blurring of vision, breathlessness and signs were smell of poison (90%), tachypnea, altered sensorium, miosis and fasciculation. POP scoring found 41% of patients in mild, 26% in moderate and 33% in sever grade of poisoning. Hospital stay ranged from 4-18 days. Complications were pulmonary Edema (PE) in 28%, Respiratory Failure (RF) 18%, Aspiration Pneumonia (AP) 15% and Intermediate Syndrome (IS) 4%. 10 died out of 42 patients who had complications and the cause of death was RF in 4, Ventricular Fibrillation (VF) 2 and IS in 2. ECG finding showed sinus tachycardia (31%), prolonged corrected QT (QTc) interval (28%), sinus bradycardia (25%), ST-T changes (17%) and Premature Ventricular Contraction (PVC) in 4% which degenerated to VF in 2%. 24 patients were Hypokalemic from which 16 developed complications.Conclusions: Similar to earlier studies we observed poisoning which was suicidal. QTc prolongation and Hypokalemia are associated with high morbidity and mortality in OPC-Poisoning.

The correlation of cardiovascular risk factors and angiographic findings with serum vitamin D levels in patients undergoing coronary angiography

Background: Studies regarding correlation of various conventional risks factors for Coronary Artery Disease (CAD) are many. Keeping in mind the scarcity of studies regarding Vit-D Deficiency (VDD), a new risk factor in CAD, present study was conducted to correlate Vit-D level with conventional risk factors and Coronary Angiography (CAG).Methods: Hundred adult patients admitted to Medicine and Cardiology undergoing CAG with suspected or established CAD were kept in study. Patients having renal, hepatic, parathyroid disease, osteomalacia and patients taking drugs interfering with Vitamin D (Vit-D) metabolism were excluded. After detailed history and thorough clinical examination, routine investigations and 25-(OH) D level was estimated. Subsequently patients underwent CAG. Statistical analysis by Mann Whitey test and Chi-square Test was done and inference was drawn.Results: 100 patients in different age groups had hypertension (HTN) in 53, diabetics mellitus (DM) 39, dyslipidemia 62, smokers 38 and family history of CAD 19. CAG showed normal coronaries in 4, Single Vessel Disease (SVD) in 30, double vessel diseases (DVD) 43 and triple vessel disease (TVD) 22. Vit-D level <20ng/ml i.e. Vit-D deficiency (VDD) in 68, 20-30ng/ml i.e. Vit-D insufficiency (VDI) in 22 and >30ng/ml (normal) in 10. VDD was profound in 51-60 and 61-70 years age groups. Statistical correlation of VDD was not significant with conventional risk factors, but statistically significant correlation of VDD was observed with dyslipidemia and CAG findings in our study.Conclusions: Like earlier studies we observed significant correlation of Vit-D levels with CAD. Though dyslipidemia was significantly correlated with VDD, correlation of other conventional risk factors like age, sex, HTN, DM, smoking and family history was not found. We conclude from this study presence of significant association of VDD with severe CAD. DVD and TVD by CAG were common with low Vit-D levels.

Brain tumor and Gliadel wafer treatment.

Glioblastoma is a rapidly progressive and extremely fatal form of brain tumor with poor prognosis. It is the most common type of primary brain tumor. Even with the most aggressive conventional treatment that comprises surgery followed by radiotherapy and chemotherapy, most patients die within a year of diagnosis. Developments in molecular and cell biology have led to better understanding of tumor development, leading to novel treatment strategies including biological therapy and immunotherapy to combat the deadly disease. Targeted drug delivery strategies to circumvent the blood-brain barrier have shown efficiency in clinical trials. Gliadel wafer is a new approach to the treatment of glioblastoma, which involves controlled release delivery of carmustine from biodegradable polymer wafers. It has shown promising results and provides a silver lining for glioblastoma patients.

Cerebral phaeohyphomycosis due to Cladophialophora bantiana.

Cladophialophora bantiana, a dematiaceous fungus and a member of the family Phaeohyphomycetes, is primarily a neurotropic fungus causing central nervous system (CNS) infection. We report a case of a well preserved, young adult male presenting with a capsuloganglionic abscess caused by C. bantiana, a rare entity. Diagnosis was made based on the mycology and histopathology findings of the aspirate from the abscess through a burr hole. The patient responded clinically to amphotericin B.

Serial pattern learning after mild head injury.

Non-verbal serial pattern learning in patients with mild brain injury was examined using a serial pattern-learning task introduced by Nissen and her colleagues. The task involves two types of pattern cycles: Simple and Complex. Each pattern cycle consists of eight acquisition trials followed by a final generation phase, i.e. the ninth trial. The subjects responded to the asterisks appearing in repetitions of a 10-element spatial sequence in each pattern cycle. Eighty subjects were taken, of which 40 subjects had mild head injury and 40 were uninjured. Prior research with this task has shown that individuals show intact performance on the indirect measure of pattern learning, but are impaired on the direct measure. The results of this study showed that mild brain injury does cause a marked disruption in the ability to learn and remember serial pattern information in both simple and complex patterns.

Unruptured intracranial aneurysms : review of natural history and recommended management strategies.

Unruptured intracranial aneurysms (UIA) are common and in the absence of evidence from randomized controlled trials, management decision can be difficult. However with the various well-planned meta-analyses, certain guidelines can be formulated for risk assessment and surgical management. UIA in patients aged more than 64 years irrespective of size, and those measuring less than 10mm in the maximum diameter should be treated conservatively, but should be evaluated yearly with MR angiography or CT angiography to look for rapid increase in size. UIA showing a diameter increase of more than 0.95mm in a year should be treated by surgery or endovascular therapy. Surgery is indicated for all UIA measuring more than 10mm in maximum diameter in patients aged less than 64 years. Endovascular therapy is indicated for UIA, which need intervention but are not surgically accessible or where surgery is contraindicated due to high-risk medical factors.

Decompressive craniectomy for stroke : indications and results.

The role of decompressive hemicraniectomy in stroke victims is gaining popularity among neuroscientists as it is found to be beneficial in malignant infarction of middle cerebral artery (MCA) territory which results in massive edema and brain swelling. Recent articles have showed that this procedure not only reduces mortality but also improves the functional outcome. We report our experience of 32 patients, who underwent this procedure. We found a good outcome in patients in whom the procedure was done early and whose pre operative Glasgow coma scale (GCS) score was 8 and above. It offered no benefit in patients whose pre operative GCS was below 5.

Facial nerve neurinoma presenting as middle cranial fossa and cerebellopontine angle mass: a case report.

Facial nerve neurinomas are rare. The tumours arising from the geniculate ganglion may grow anteriorly and superiorly and present as a mass in the middle cranial fossa. Only a few cases of facial nerve neurinomas presenting as middle cranial fossa mass have so far been reported. These tumours present with either long standing or intermittent facial palsy along with cerebellopontine angle syndrome.

CSF rhinorrhoea from unusual site : report of two cases.

CSF rhinorrhoea is associated with high morbidity and mortality. Bone and dural defects may result from trauma or enlarging 'pitholes' or breach in lateral recess of sphenoid sinus. Unless surgically corrected, they tend to cause meningitis and rhinorrhoea. Unusually delayed rhinorrhoea is a diagnostic problem.