BRAF V600E mutation detection in hairy cell leukemia-utility of archival DNA from bone marrow aspirate/imprint smear and amplification refractory mutation system.

BRAF V600E is a disease defining mutation for hairy cell leukemia (HCL), which helps in its diagnosis and differentiation from morphologically similar splenic marginal zone lymphoma (SMZL) and HCL-variant (HCL-v). Forty eight cases:HCL(n = 34), SMZL(n = 11) and HCL-v(n = 3) were included. Of these, 32 were retrospective and 16 were prospective. DNA was extracted, in retrospective cases from cells obtained by smears from bone marrow aspirate/trephine imprint (BMA/BMTx) slides, and in prospective cases from peripheral blood (PB)/BMA samples. BRAF V600E mutation testing was done using ARMS-PCR. BRAF V600E mutation was positive in all HCL and negative in all SMZL and HCL-v cases. DNA extracted from BMA/BMTx slides gave results comparable to DNA extracted from PB/BMA samples. Median age of presentation for HCL (53 years) and SMZL (56 years) were quite similar, however, HCL-v (71 years) cases presented at an older age. Statistically significant differences between the three groups were seen for total leucocyte, platelet, absolute lymphocyte and monocyte counts, bone marrow-infiltration pattern, reticulin fibrosis, and an expression of CD11c, CD25, CD103, CD123, and CD200. The use of BMA/BMTx smears for DNA extraction was found to be a useful alternative to DNA extraction from formalin-fixed paraffin-embedded biopsy sections. ARMS-PCR is an efficient and specific technique to detect BRAF V600E mutation in HCL patients.

Fine needle aspiration cytology of primary and metastatic gastrointestinal stromal tumour.

AIM: To explore the cytological spectrum of the gastrointestinal stromal tumour (GIST) including its metastatic sites. MATERIAL AND METHODS: A total of 42 patients (45 sites) diagnosed with GIST or its metastases on fine needle aspiration cytology were studied over a period of 5 years. May-Grünwald Giemsa- and haematoxylin and eosin-stained smears were reviewed and analysed for the cytomorphological spectrum of GIST. RESULTS: Primary GIST alone was seen in 24 cases, E-GIST in eight cases and metastasis in 11 cases (one patient showing metastasis at two distinct sites), whereas concurrent primary and metastatic lesions were noted in two cases. Amongst primary sites, the most commonly affected location was stomach (n = 22), followed by ileum (n = 2), duodenum (n = 1) and rectum (n = 1). Extra-GIST was seen in retroperitoneum and pelvis (n = 3 each), omentum and mediastinum (n = 1 each). Fine needle aspiration cytology was done from 11 metastatic sites of GIST which included liver, gall bladder fossa, chest wall, and thigh. The classic spindle cell arrangement was the predominant cytological pattern. About 8.8% cases showed predominant epithelioid cell morphology and 15.5% cases had a mixed cytomorphology comprising of both spindle cell and epithelioid cell patterns. Nuclear pseudoinclusions, perinuclear vacuoles and multinucleation were seen in four cases. Immunocytochemistry on cell-block sections for confirmation was performed in 18 cases and all these cases showed strong c-KIT positivity. CONCLUSION: In this largest case series of cytomorphological diagnosis of GIST, we describe the cytomorphology and immunocytochemistry of primary and metastatic GIST. GISTs with predominant epithelioid cell morphology may pose a diagnostic dilemma therefore in all suspected cases of GIST, immunocytochemistry for c-KIT and/or DOG1 should be employed on cell-block preparations to confirm the diagnosis of GIST.

Comparative Evaluation of Cost-Effectiveness, Clinical and Microbiological Parameters of Systemic Antibiotics Versus Local Drug Delivery in Aggressive Periodontitis.

BACKGROUND: Periodontitis is a biofilm-associated inflammatory disease of the periodontium in which microbial component contributes to its initiation that mainly includes chronic periodontitis and aggressive periodontitis (AgP). AIM: The present study aimed to ascertain a cost-effective treatment approach in AgP with minimal side effects by comparing systemic antibiotics (SA) and local drug delivery (LDD) as an adjunct to scaling and root planning (SRP). Furthermore, the objectives were to analyze its effect on clinical and microbial parameters. MATERIALS AND METHODS: The participants were randomly divided into two groups A and B with eleven participants each. Group A was administered with Chlosite Gel [LDD] whereas group B with amoxicillin (AMX) 500 mg + metronidazole (MTZ) 400 mg [SA] thrice daily for 14 days. Clinical parameters such as plaque index (PI), gingival index (GI), clinical attachment level (CAL), and pocket probing depth (PPD)] were recorded at baseline and three months postoperatively. Microbiological parameters i.e Aggregatibacter actinomycetemcomitans (Aa) and Porphyromonas gingivalis (Pg) counts were also measured at baseline and after three months. RESULTS: Statistically significant difference was observed in all clinical parameters i.e. PI, GI, CAL, and PPD after three months follow-up in groups A and B. Reduced bacterial load i.e. Aa and Pg was significant at three months in both the groups. However, on comparing the total expenditure of both the groups, group A ranged from 920-1480 ₹ while in group B it ranged from 330-360 ₹. Hence, lower expenditure in group B (3.7 fold) compared to group A was evident. CONCLUSIONS: Cost-effective approach recommended in the present study is mainly to promote awareness among periodontal patients in the public and private sectors, who refuse to get dental treatment due to fear of high expenditure. This can be attained by incorporating SA as an alternative to LDD in AgP patients.

Mechanical Thrombectomy in Embolic Cardiac Myxoma: Case Report and Literature Review.

Myxomas are the most common cardiac tumors and present clinically with cardiac manifestations, systemic constitutional symptoms, and embolic events. Posterior circulation involvement occurs in approximately 20 percent of cerebral ischemic events.The endovascular technique is an established life-saving therapy for eligible patients upto 24 hours from symptom onset. However, the role of endovascular management in embolic atrial myxoma remains unknown with no international consensus guidelines for the management of stroke in such patient population. Here, we present a case report of an embolic posterior circulation stroke in a young female treated with mechanical thrombectomy at 23 hours from symptom onset. To the best of our knowledge, this is the first thrombectomy case in posterior circulation with embolism from myxoma. Further workup confirmed an atrial myxoma which was resected. We also review the previous cases with mechanical thrombectomy done in such cases.

Molecular Testing for BRAFV600E and RAS Mutations from Cytoscrapes of Thyroid Fine Needle Aspirates: A Single-Center Pilot Study.

CONTEXT AND AIM: Molecular testing of thyroid FNA has been advocated in the indeterminate categories of The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC) 2018. The utility of cytoscrapes of thyroid FNA samples for BRAF V600E and RAS mutations was evaluated in this pilot study. METHODS AND MATERIALS: Thyroid FNA samples between 2015 and 2018 from TBSRTC categories 3-6 were included. DNA was extracted from one to two representative smears (cytoscrape). Real-time PCR for BRAF V600E and RAS (KRAS, NRAS, and HRAS) gene mutations was performed. Histopathology correlation was available in 44 cases. STATISTICAL METHODS: Chi-square test and calculation of sensitivity, specificity, and positive/negative predictive values were performed. RESULTS: A total of 73 thyroid FNA cases and 11 nodal metastases of papillary thyroid carcinoma (PTC) were evaluated. The DNA yield ranged from 1.9 to 666 ng/µl (mean 128 ng/µl) in 80 cases and was insufficient in four cases. Overall, mutations were seen in 45 (56.25%) cases with BRAF V600E, NRAS, HRAS, and KRAS in 21 (46.7%), 19 (42.2%), 4, and 1 cases, respectively. BRAF V600E mutation was seen in PTC (11/18, 61%), nodal PTC metastases (5/10, 50%), and occasionally in TBSRTC category 3 (1/18, 5.5%). NRAS mutations were seen across all categories and were maximum in the AUS/FLUS group (6/18, 33%). BRAF V600E /RAS testing had an overall sensitivity, specificity, and positive and negative predictive values of 61.7%, 80%, 91.3%, and 38%, respectively, for the detection of malignancy. In indeterminate thyroid nodules, the sensitivity, specificity, PPV, and NPV were 56.2%, 80%, 81.8%, and 53.3%, respectively. CONCLUSION: BRAF V600E/RAS mutation testing from cytoscrapes are useful as a rule-in test for indeterminate thyroid nodules and provide molecular confirmation in nodal metastases of PTC.