Use of cytology fluid samples for predictive biomarker testing in lung cancer patients.

OBJECTIVE: To provide a method of directly using cytology fluid samples for predictive biomarker testing in lung cancer patients and to determine the efficacy of a variety of fluid sample types. METHOD: A review of our in-house data from a range of cytology samples including endobronchial ultrasound (EBUS) fine-needle aspirate (FNA) needle washings (NW) and serous effusions tested on the Biocartis Idylla platform. All fluid samples were originally tested using Sanger sequencing. RESULTS: Using our method for fluid samples all of our cytology samples tested for epithelial growth factor receptor (EGFR) yielded valid results on this platform and all variant cases identified. The data showed serous fluids provided the best quality DNA, and variant genotype reports were obtained within 150 minutes. CONCLUSION: Cytology fluid samples can be used for predictive biomarker testing for lung cancer patients to provide in-house results with all fluids providing good-quality DNA.

Pretracheal ectopic thymoma: A diagnostic challenge in endobronchial ultrasound-guided transbronchial needle aspiration cytology.

Ectopic thymomas (ETs) are rare thymic neoplasms that arise from atypical anatomical sites and present a diagnostic challenge for clinicians as they can be mistaken for other pathological entities on fine needle aspiration (FNA) cytology.

Pleural malignant mesothelioma with metastatic deposition in the tongue.

A 70-year-old man with epithelioid malignant pleural mesothelioma was referred to the maxillofacial surgery department with a painful lesion on the lateral border of his tongue which interfered with swallowing. Physical examination demonstrated a 3 cm firm, non-ulcerated mass on the left lateral tongue, histologically shown to be metastatic epithelioid mesothelioma. MRI of the neck and CT scan of the thorax, abdomen and pelvis scans also demonstrated progressive local and metastatic disease. The patient was not felt to be fit for further treatment. More than 90% of oral cavity cancers are squamous cell carcinoma. All reported cases of metastasis of malignant mesothelioma to the tongue have been of the epithelioid subtype with 10 cases reported in the literature to date. Diagnosis is important as tongue lesions can significantly impact on quality of life and synchronous tongue primaries should be ruled out. Tongue lesions may herald metastases to other extrapleural locations.

Use of the Idylla EGFR Mutation Test for Variant Detection in Non-Small Cell Lung Cancer Samples.

OBJECTIVES: Stratified management of patients with non-small cell lung carcinoma (NSCLC) through epithelial growth factor receptor (EGFR) variant analysis has become standard clinical practice. The Idylla system is a fully compliant European in vitro diagnostics device, a fully automated platform designed to rapidly genotype formalin-fixed paraffin-embedded tissue samples. This retrospective study aims to validate the Idylla EGFR Mutation Test for use with extracted DNA from known NSCLC samples. METHODS: In this study, 20 µL of archival extracted DNA was placed directly inside the Idylla EGFR assay single-use cartridge. Idylla results were compared with the original Sanger sequencing reference method. RESULTS: The Idylla EGFR Mutation Test yielded valid results for all samples tested, confirming the variants identified by the reference method that lay within the Idylla target range. No false-positive cases were noted with the Idylla assay. Variant genotype reports were obtained within 150 minutes. CONCLUSIONS: The Idylla EGFR assay is sensitive for extracted DNA and can be reliably applied to cytologic specimens, enabling its implementation as an ancillary first-line test for patients with NSCLC.

Evolution of a rapid onsite evaluation (ROSE) service for endobronchial ultrasound guided (EBUS) fine needle aspiration (FNA) cytology in a UK Hospital: A 7 year audit.

OBJECTIVE: Endobronchial ultrasound fine needle aspiration (EBUS FNA) is a well-established procedure for the diagnosis and staging of lung cancer. We review our provision of this service at the Royal Devon and Exeter NHS Foundation Trust and the role of rapid onsite evaluation (ROSE) with the increasing demand for molecular markers in this era of personalized medicine. METHODS: A review of the changes in the Endoscopy clinic over the 7 years from the introduction of EBUS at the end of 2010 until 2017 was carried out. This included the availability of material obtained for diagnosis, accurate subtyping, and molecular testing. We also assessed the success of molecular genetics DNA techniques from EBUS material versus formalin fixed paraffin embedded tissue (FFPE). RESULTS: A total of 1218 EBUS cases with ROSE were reported between 2011 and 2017 Percentage diagnostic rates were calculated as 83, 82, 84, 92, 93, 94, and 92 for 2011, 2012, 2013, 2014, 2015, 2016, and 2017, respectively. Availability of material for immunocytochemistry ranged from 86 to 100% over the 7 years. Molecular testing was successfully performed for EGFR in 89-100% of requested cases and ALK testing in 87-100% of requested cases. EBUS sourced material gave on average twice the amount of DNA and fewer amplicon repeats per patient compared to FFPE material. CONCLUSION: ROSE at EBUS FNA provides access to suitable material for molecular testing with increased yields in the form of needle washings for EGFR with FFPE materials for ALK and PDL1 testing.

A case report of squamous cell carcinoma in a suprapubic urinary catheter tract: surgical excision and simultaneous colostomy formation.

Squamous cell carcinoma (SCC) arising from a suprapubic cystostomy tract is a rare complication of long-term suprapubic catheterization (SPC). A 53-year-old man with paraplegia secondary to spina bifida presented with a painful granulomatous lesion around his SPC site that was being treated with silver nitrate cauterization in the community. Consequently, he developed a sacral pressure sore due to reduced mobility from the pain. He also had increasing difficulties with defaecation secondary to his spina bifida. His sacral pressure sore was secondary to a cryptoglandular fistula with coccygeal osteomylelitis. Post-operative pathology revealed infiltrative SCC involving full thickness of the specimen from skin to the bladder wall with clear surgical margins. We describe the first case requiring a simultaneous suprapubic tract SCC excision and colostomy formation. We recommend early investigation of lesions arising from a long-term suprapubic tract especially in patients with spinal cord injuries or congenital defects.

Clinically unsuspected bilateral malignant pheochromocytoma.

Pheochromocytoma is an uncommon tumor and occurrence of bilateral malignant tumor is extremely rare. Labile hypertension is the common presenting feature of pheochromocytomas. Herein we describe one such case ofbilateral malignant pheochromocytoma, which was clinically silent except for pain and mass in hypochondrium. Histopathology showed features of malignancy along with hyaline globules, which are less described with malignant tumors. Electron microscopy showed neurosecretory granules in tumor cells.

Dying from cardiac tamponade.

BACKGROUND: To determine the causes of cardiac tamponade (CT), focussing especially on haemopericardium (HP), as a terminal mode of death, within a 430,000 rural English population. METHODS: Our hospital mortuary register and, all postmortem reports between 1995 and 2004 inclusive, were interrogated for patients dying of CT or HP. The causes of CT/HP and selected morphological characteristics were then determined. RESULTS: 14,368 postmortems were performed in this period: of these, 461 patients died of CT. Three cases were due to non-haemorrhagic pericardial effusion. HP accounted for the remaining 458 cases of which, five were post-traumatic, 311 followed rupture of an acute myocardial infarction (RAMI), 138 after intra-pericardial rupture of dissecting ascending aortic aneurysms (RD3A) and four were due to miscellaneous causes.HP was more commonly due to RAMI. Men tended to die from RAMI or RD3A earlier than women. RAMI or RD3A were commoner in men <70 yrs, but more frequent in women after this.Two thirds of RAMI were associated with coronary artery thrombosis. Anterior free wall rupture was commonest overall, and in women, but posterior free wall rupture was commoner in men.The volume of intrapericardial blood in RAMI (mean = 440 ml) and RD3A (mean = 498 ml) varied between 150 and 1000 ml: intrapericardial blood volume was greater in men than in women dying from either RAMI or RD3A. CONCLUSION: At postmortem, CT is most often related to HP, attributable to either RAMI or intrapericardial RD3A. Post-traumatic and other causes of CT are infrequent.

Pigmented vagal paraganglioma.

Paragangliomas are uncommon and those arising from the vagal trunk are rarer. Pigmented extra-adrenal paragangliomas are still rarer and reported sites of occurrence are the uterus, spine, retroperitoneum, bladder, mediastinum and orbit. The presence of abundant pigment in a cervical paraganglion has not been reported previously. We report one such unusual case of pigmented vagal paraganglioma that arose from the vagal trunk below the nodose ganglion, had massive central necrosis and showed hypovascularity on angiography. The unusual features and difficulties in the diagnosis of such cases are discussed.

Papillary carcinoma thyroid presenting as a choroidal metastasis. Report of a case and brief review of literature.

A patient of papillary thyroid carcinoma, who initially presented with a choroidal metastasis and secondary intractable glaucoma is reported. The orange-coloured tumour posed a diagnostic difficulty, until histopathology of the enucleated eye confirmed the metastasis from the thyroid. The initial presentation of distant metastasis in patients with thyroid cancer is rare.

Flow cytometric characterization of phenotype, DNA indices and p53 gene expression in 55 cases of acute leukemia.

OBJECTIVE: To characterize the phenotype of acute leukemia cases using flow cytometry, to detect mixed lineage cases and to use DNA index determination, including S-phase fraction (SPF) and p53 detection, to find if there was any correlation of SPF and p53 expression with outcome. STUDY DESIGN: Fifty-five cases of acute leukemia were enrolled in this study. A complete hemogram and routine bone marrow examination, including cytochemistry, was done. Mycloperoxidase-negative cases were evaluated on a flow cytometer using monoclonal antibodies. DNA indices were determined by flow cytometry in all cases, and p53 was detected immunohistochemically using the alkaline phosphatase/antialkaline phosphatase technique. RESULTS: Acute myeloblastic leukemia (AML) was diagnosed in 32 cases; acute lymphoblastic leukemia (ALL) was diagnosed in 18 (14 B lineage and 4 T line age). Four cases showed mixed lineage leukemia, and undifferentiated acute leukemia was diagnosed in one case. The mean/range of SPF for these groups were 3.76/0.33-6.91, 6.25/0.15-21.4, 2.89/0.35-10.64, 2.60/0.72-6.94 and 7.34, respectively. Aneuploidy was detected in two cases of B-lineage ALL and tetraploidy in a case of AML-M7, while all others were diploid p53. Was detected in 6 of 55 cases (10.90%). Follow-up was available for 24 patients. Five patients relapsed, and four had B-cell type ALL and were diploid and expressed no p53 gene. SPF% did not show any correlation with outcome. CONCLUSION: These data suggest that within acute leukemia subtypes, there is a wide variation in SPF. SPF does not seem to correlate with outcome. Immunophenotyping is essential to determine the lineage in myeloperoxidase-negative cases. It is perhaps the only way to diagnose mixed lineage leukemia and aberrant expression of markers presently. The p53 gene was detected less frequently. However, more studies are required from different centers with longer follow-up to evaluate prognostic significance.

Mean nuclear volume in cervical intraepithelial neoplasia and carcinoma.

OBJECTIVE: To correlate three-dimensional nuclear size (mean nuclear volume) estimated by the stereologic intercept methodfor objective classification of cervical intraepithelial neoplasia (CIN) and carcinoma. STUDY DESIGN: In this retrospective study a total number of 29 CIN cases (8 cases of CIN 1, 10 cases of CIN 2 and 11 cases of CIN 3) and 10 cervical squamous cell carcinoma cases were selected. Mean nuclear volume (MNV) of all cases was measured with an image cytometer (Leica, Cambridge, England) using Quantimet 600 software (Leica). Nuclear point resection method was adopted to measure nuclear volume. Mean intercepted diameter of at least 50 nuclei was measured randomly. MNV was correlated with the histologic grade and diagnosis. RESULTS: MNV of CIN 1, 2, 3 and carcinoma cases was 291.72, 403.33, 711.45 and 893 microm3, respectively. ANOVA test results showed that MNV of CIN 1 and 2 was significantly lower than that of CIN 3 and invasive carcinoma (P < .000). MNV of CIN 3 was also significantly lower than that of carcinoma cases (P <.05). CONCLUSION: The findings suggest that estimates of MNV on conventional histopathology slides provide objective and useful criteria for relatively subjective histopathologic grading.

Myofibroblastoma of the male breast: a diagnostic problem on fine-needle aspiration cytology.

Myofibroblastoma is an uncommon neoplasm of the male breast. Herein, we describe the cytologic features seen in the fine-needle aspirate of a 45-year-old man. The smears were cellular with intimate association of tumor cells with extracellular matrix material. The cells were spindle to polygonal and were without significant atypia. Numerous mast cells were observed. Nuclear grooving was present only occasionally, although this was conspicuous histologically. The presence of hyaline bands in between tumor cells, another interesting feature, was appreciated retrospectively. This neoplasm was initially misinterpreted as a malignant soft tissue tumor. Awareness of the cytologic features coupled with mammography should prevent a misdiagnosis of this tumor.