Medullary thyroid carcinoma diagnosed with liquid-based cytology and immunocytochemistry.

Medullary thyroid carcinoma (MTC) is a rare neuroendocrine malignancy that arises from the parafollicular cells (C cells) secreting calcitonin. This study summarizes our experience in the diagnosis of MTC with ultrasound-guided thyroid FNA, subsequently processed with liquid-based cytology (LBC) and immunocytochemistry (ICC). We searched our laboratory archives for thyroid FNA cases with an interpretation of positive or suspicious for MTC, during the period 2004-2018. A total of 20 cases (18 thyroid FNAs; two lymph node FNAs) were selected and included in this study. These displayed high cellularity and a discohesive pattern, with a few loose syncytial groups. There was some variation in the cell size and shape both across and within our cases. Most MTCs (n = 15) exhibited a predominant plasmacytoid/epithelioid cell morphology, whereas five of our cases showed a spindle cell pattern. Of interest, none of eight MTC microcarcinomas (≤1 cm) showed a spindle cell morphology. Amyloid was found in 11/20 cases (55%), while binucleation/multinucleation in 17/20 (85%), and nuclear pseudoinclusions in 3/20 MTC cases (15%). Nuclei exhibited a granular, "salt and pepper" chromatin in all cases. ICC was performed in 18/20 cases (90%). Calcitonin, CEA, TTF1, and Chromogranin were positive wherever applied, whereas thyroglobulin and CK19 were negative. In conclusion, ultrasound-guided thyroid FNA - processed with LBC and ICC - is a reliable modality to detect MTC preoperatively, facilitating the management of such patients.

Immunity onset alters plant chromatin and utilizes EDA16 to regulate oxidative homeostasis.

Perception of microbes by plants leads to dynamic reprogramming of the transcriptome, which is essential for plant health. The appropriate amplitude of this transcriptional response can be regulated at multiple levels, including chromatin. However, the mechanisms underlying the interplay between chromatin remodeling and transcription dynamics upon activation of plant immunity remain poorly understood. Here, we present evidence that activation of plant immunity by bacteria leads to nucleosome repositioning, which correlates with altered transcription. Nucleosome remodeling follows distinct patterns of nucleosome repositioning at different loci. Using a reverse genetic screen, we identify multiple chromatin remodeling ATPases with previously undescribed roles in immunity, including EMBRYO SAC DEVELOPMENT ARREST 16, EDA16. Functional characterization of the immune-inducible chromatin remodeling ATPase EDA16 revealed a mechanism to negatively regulate immunity activation and limit changes in redox homeostasis. Our transcriptomic data combined with MNase-seq data for EDA16 functional knock-out and over-expressor mutants show that EDA16 selectively regulates a defined subset of genes involved in redox signaling through nucleosome repositioning. Thus, collectively, chromatin remodeling ATPases fine-tune immune responses and provide a previously uncharacterized mechanism of immune regulation.

Radial Basis Function Artificial Neural Network for the Investigation of Thyroid Cytological Lesions.

OBJECTIVE: This study investigates the potential of an artificial intelligence (AI) methodology, the radial basis function (RBF) artificial neural network (ANN), in the evaluation of thyroid lesions. Study Design. The study was performed on 447 patients who had both cytological and histological evaluation in agreement. Cytological specimens were prepared using liquid-based cytology, and the histological result was based on subsequent surgical samples. Each specimen was digitized; on these images, nuclear morphology features were measured by the use of an image analysis system. The extracted measurements (41,324 nuclei) were separated into two sets: the training set that was used to create the RBF ANN and the test set that was used to evaluate the RBF performance. The system aimed to predict the histological status as benign or malignant. RESULTS: The RBF ANN obtained in the training set has sensitivity 82.5%, specificity 94.6%, and overall accuracy 90.3%, while in the test set, these indices were 81.4%, 90.0%, and 86.9%, respectively. Algorithm was used to classify patients on the basis of the RBF ANN, the overall sensitivity was 95.0%, the specificity was 95.5%, and no statistically significant difference was observed. CONCLUSION: AI techniques and especially ANNs, only in the recent years, have been studied extensively. The proposed approach is promising to avoid misdiagnoses and assists the everyday practice of the cytopathology. The major drawback in this approach is the automation of a procedure to accurately detect and measure cell nuclei from the digitized images.

GCN5 modulates salicylic acid homeostasis by regulating H3K14ac levels at the 5' and 3' ends of its target genes.

The modification of histones by acetyl groups has a key role in the regulation of chromatin structure and transcription. The Arabidopsis thaliana histone acetyltransferase GCN5 regulates histone modifications as part of the Spt-Ada-Gcn5 Acetyltransferase (SAGA) transcriptional coactivator complex. GCN5 was previously shown to acetylate lysine 14 of histone 3 (H3K14ac) in the promoter regions of its target genes even though GCN5 binding did not systematically correlate with gene activation. Here, we explored the mechanism through which GCN5 controls transcription. First, we fine-mapped its GCN5 binding sites genome-wide and then used several global methodologies (ATAC-seq, ChIP-seq and RNA-seq) to assess the effect of GCN5 loss-of-function on the expression and epigenetic regulation of its target genes. These analyses provided evidence that GCN5 has a dual role in the regulation of H3K14ac levels in their 5' and 3' ends of its target genes. While the gcn5 mutation led to a genome-wide decrease of H3K14ac in the 5' end of the GCN5 down-regulated targets, it also led to an increase of H3K14ac in the 3' ends of GCN5 up-regulated targets. Furthermore, genome-wide changes in H3K14ac levels in the gcn5 mutant correlated with changes in H3K9ac at both 5' and 3' ends, providing evidence for a molecular link between the depositions of these two histone modifications. To understand the biological relevance of these regulations, we showed that GCN5 participates in the responses to biotic stress by repressing salicylic acid (SA) accumulation and SA-mediated immunity, highlighting the role of this protein in the regulation of the crosstalk between diverse developmental and stress-responsive physiological programs. Hence, our results demonstrate that GCN5, through the modulation of H3K14ac levels on its targets, controls the balance between biotic and abiotic stress responses and is a master regulator of plant-environmental interactions.

A Rare Pancreatic Tail Metastasis from Squamous Cell Lung Carcinoma Diagnosed by EUS-FNB and a Small Review of the Literature.

Differential diagnosis of pancreatic lesions is really challenging, especially when the patient is diagnosed with primary cancer at another site. In this case report, we managed to histologically confirm pancreatic metastasis from squamous cell lung carcinoma, which is a very rare entity, using endoscopic ultrasound fine needle biopsy.

O diagnóstico diferencial de lesões pancreáticas é desafiante, especialmente quando o doente é diagnosticado com cancro primário noutro local. Neste caso clínico foi possível confirmar histologicamente o diagnóstico de metástase pancreática de carcinoma de células escamosas do pulmão, que se trata de uma entidade muito rara, através de punção com agulha fina ecoguiada por ecoendoscopia.

Disseminated metastatic cutaneous melanoma to pancreas and upper gastrointestinal tract diagnosed by endoscopic ultrasound: an unusual case.

There is a wide variety of pancreatic neoplasms identified, but the great majority of them are of primary origin. Metastatic disease in the pancreatic parenchyma is quite rare (2-5% of pancreatic malignancies) and most often is quite difficult to differentiate from other primary lesions. Most of the imaging studies fail to give certain discriminating features for metastatic pancreatic neoplasms, contrary to endoscopic ultrasound and tissue sampling, which can provide an accurate diagnosis. In this report, we present a case of a male middle aged man who was admitted to our hospital with painless jaundice and finally was diagnosed with a cutaneous scalp melanoma dispersedly metastasized to the pancreas and upper gastrointestinal tract (stomach and duodenum).

Large adrenal cyst masquerading as a pancreatic cystic tumor: a rare diagnosis based on endoscopic ultrasound.

Adrenal cystic lesions constitute a rare finding and usually are diagnosed incidentally as retroperitoneal cysts during imaging studies. A major issue, especially for the large-sized ones, is their preoperative diagnosis and management, as imaging modalities often fail to detect their exact origin prior to surgery. We report a case of a big adrenal cyst that was mistakenly diagnosed and followed up as pancreatic cystic tail tumor. Our thorough workup, which included endoscopic ultrasound (EUS), managed to delineate an adrenal gland-originated cyst and altered further management of the patient. According to our knowledge, this is the first report in literature in which EUS proved its superiority over other imaging studies in clarifying the origin of an ambiguous big retroperitoneal cyst. We believe that for cases of large retroperitoneal cystic lesions, EUS consists the optimal diagnostic and sampling tool.

Application of Immunocytochemistry on Cell Block Sections for the Investigation of Thyroid Lesions.

OBJECTIVE: To investigate the potential of Classification and Regression Trees (CARTs) for the diagnosis of thyroid lesions based on cell block immunocytochemistry and cytological outcome. STUDY DESIGN: A total of 956 histologically confirmed cases (673 benign and 283 malignant) from patients with thyroid nodules were prepared via liquid-based cytology and evaluated; 4 additional slides were stained for cytokeratin 19 (CK-19), galectin 3 (Gal-3), Hector Battifora mesothelial cell 1 (HBME-1), and thyroglobulin. On the basis of immunocytochemistry and the cytological diagnosis, a CART algorithm was constructed and used for evaluation. RESULTS: The major important factors contributing to the diagnostic CART model were: cytological outcome, CK-19, Gal-3, and HBME-1. The sensitivity and specificity of the cytological diagnosis were 96.27% and 88.26%, respectively (cut-off: category 3 of The Bethesda System [TBS-3]). The introduction of immunocytochemistry and the CART model increased the sensitivity and specificity to 98.88% and 99.11%, respectively. CK-19 presented the best performance for discriminating papillary thyroid carcinomas, followed by HBME-1 and Gal-3. In the TBS-2 cases, CK-19 and, subsequently, Gal-3 were important immunocytochemistry markers. Ultimately, CK-19 and HBME-1 on TBS-5 or TBS-6 cases demonstrated the best results. CONCLUSIONS: The hierarchical structure of the CART model provides a diagnostic algorithm linked with the risk of malignancy at every step of the procedure. It also provides guidance on the use of ancillary examinations as it goes by simple, human understandable rules.

Classification and regression trees for the evaluation of thyroid cytomorphological characteristics: A study based on liquid based cytology specimens from thyroid fine needle aspirations.

BACKGROUND: This study investigates the potential of classification and regression trees (CARTs) for the evaluation of thyroid lesions. METHODS: The study was performed on 521, histologically confirmed cytological specimens prepared via liquid based cytology. For each specimen, contextual and cellular morphology features were recorded by experienced cytopathologists, as described in everyday cytological practice and The Bethesda System (TBS); these features were subsequently used to construct two CART models, viz. CART-C for the prediction of the cytological diagnosis (according to TBS) and CART-H for the prediction of the histological diagnosis (hereby expressed as either benign or malignant). RESULTS: CART-C had no statistically significant performance from the cytologists' evaluations and CART-H had a very good predictive performance for the histological status. CONCLUSION: CARTs provide a methodological framework capable for data mining and knowledge extraction. They created simple human understandable rules and classification algorithms that may assist cytopathologists towards decisions based on classification steps, each one linked with a specific risk and moreover by applying cytomorphological characteristics in hierarchical order according to their importance. The two CARTs may be a useful tool for the training of nonexperienced cytopathologists; moreover, they may act as ancillary methods to avoid misdiagnoses and assist quality assurance procedures in the everyday practice of the cytopathology laboratory.

Fine needle aspiration cytology diagnosis of primary renal lymphoma involving the pleura: a case report.

BACKGROUND: Primary renal lymphoma is a rare disease (< 1% of kidney lesions). We present a case of renal large B-cell type non-Hodgkin's lymphoma (NHL) with right sided pleural involvement. CASE: A 70-year-old man was admitted with persistent, painless, macroscopic hematuria for 1 month. Ultrasound examination, abdominal computed tomography and magnetic resonance imaging techniques revealed a large tumor in the right kidney extending in the perirenal area. The patient underwent a radical nephrectomy for suggested renal cell carcinoma. He developed thoracic pain and pleural effusion in the 10 days after surgery. The pleural fluid was cytologically processed using conventional and ThinPrep (Cytyc Corporation, Boxborough, Massachusetts, U.S.A.) cytopreparatory techniques, slides were Papanicolaou and Giemsa stained, and immunocytochemistry was performed on the ThinPrep slides. The cytologic examination of the fluid specimen revealed a highly cellular smear composed of dispersed neoplastic cells of intermediate and large size. Immunocytochemically, the neoplastic cells were: CD45 (LCA) (+), CD20 (+), CK7 (-), CK20 (-), NSE (-), CD45 RO (UCHL-1) (-) and CD30 (-). On cytomorphologic and immunocytologic examination, the specimen fulfilled the criteria of a large B-cell type NHL. Histologic evaluation of the nephrectomy specimen revealed an infiltrating, diffuse large cell renal NHL, B-cell type, of immunoblastic and centroblastic morphology. This NHL was considered a renal primary because no peripheral lymphadenopathy or hepatosplenomegaly was revealed by the imaging techniques. CONCLUSION: Cytomorphologic and immunocytologic examination revealed the typical features of a renal large B-cell type NHL in a case with pleural involvement.