Hyang Mi Ko

Diagnosis and subclassification of lymphomas and non-neoplastic lesions involving mediastinal lymph nodes using endobronchial ultrasound-guided transbronchial needle aspiration

Introduction: The value of endobronchial ultrasound‐guided transbronchial needle aspiration (EBUS‐TBNA) has been established for staging mediastinal lymph nodes in lung carcinoma patients with radiologically enlarged lymph nodes, but its utility for evaluation of primary lymph node disorders is not well defined. The objective of this study was to evaluate the usefulness of EBUS‐TBNA with on‐site assessment and triage of sample for multiple ancillary techniques, for the diagnosis and subclassification of lymphomas and non‐neoplastic lesions involving mediastinal lymph nodes. Methods: One hundred and twenty consecutive patients who underwent EBUS‐TBNA between January 2008 and August 2009 were reviewed. The final cytological diagnosis was based on air‐dried Romanowsky and alcohol‐fixed Papanicolaou stained direct smears, immunohistochemistry, immunophenotyping, and fluorescence in situ hybridization (FISH). Results: A total of 38 cases were included in this study consisting of eight reactive lymphoid hyperplasia, 20 granulomatous lymphadenitis (17 non‐necrotizing and 3 necrotizing granulomatous inflammations), 3 Hodgkin lymphomas and 7 non‐Hodgkin lymphomas (1 small lymphocytic lymphoma (SLL), 1 SLL with scattered Reed‐Sternberg cells, 1 marginal zone lymphoma, and 4 large B cell lymphomas). Cultures performed in 13 cases were negative for AFB and fungi. Immunophenotyping and immunohistochemistry for MIB1 in six cases, and FISH in five cases provided necessary information for subclassification. Conclusions: EBUS‐TBNA is a minimally invasive procedure which provides sufficient sample for definitive primary diagnosis and classification of malignant lymphoma and granulomatous inflammation in patients with mediastinal lymphadenopathy. Rapid on‐site specimen assessment is invaluable for appropriate assignment of sample to ancillary studies. Diagn. Cytopathol. 2013;41:1023–1030.

“The petals and thorns” of ROSE (rapid on‐site evaluation)

The small samples obtained by these procedures can be used for diagnosis based on morphologic criteria alone and also, in many instances, for special ancillary studies, including molecular analysis. The inherent challenges involved in handling and processing limited samples in such a way as to allow multiple studies have triggered changes in the preanalytical phase and generated new protocols to maximize tissue/cell retrieval. 5 ROSE has been advocated as an effective way to ensure that samples are handled properly for morphological analysis, and that they meet all preanalytical requirements for specific diagnostic tests. Emerging novel molecular diagnostic technologies in current pathology practice have reinforced the essential role of ROSE in allowing pathologists to accurately divide diagnostic cytological material into small aliquots. This procedure ensures that sufficient quantities of cells of adequate quality are obtained to permit a complete diagnostic workup. Ultimately, this will translate into an appropriate treatment plan. For CT-guided and endoscopicguided procedures, ROSE has become increasingly more relevant because of the higher (although still low) rate of complications and the intrinsic risks of these procedures when compared with FNA of superficial lesions. A recent trial has concluded that ROSE of transbronchial aspirates from hilar and mediastinal lymph nodes enables clinicians to avoid additional biopsy without a loss in the diagnostic yield and reduces the complication rate of bronchoscopy. 6

Docetaxel-loaded thermoresponsive conjugated linoleic acid-incorporated poloxamer hydrogel for the suppression of peritoneal metastasis of gastric cancer.

We evaluated the potential of a thermoresponsive hydrogel consisting of conjugated linoleic acid-coupled Pluronic F-127 (Plu-CLA) as a controlled release, intraperitoneal delivery system for docetaxel with the aim of treating peritoneal dissemination of gastric cancer. Previously, we established a peritoneal metastasis model that involves the injection of BALB/c mice with TMK1 human gastric cancer cells. One week after the TMK1 cells were injected, the mice were injected intraperitoneally with docetaxel alone or docetaxel-loaded Plu-CLA. Tumor progression and response to therapy were monitored by micro-positron emission tomography. The total number of peritoneal tumors and the ascites volume were also measured. Compared with docetaxel alone, the combination of docetaxel and Plu-CLA (docetaxel-Plu-CLA) significantly and synergistically reduced tumor cell survival. Docetaxel-Plu-CLA showed excellent anti-tumor activity, inducing apoptosis more potently than docetaxel alone. Docetaxel-Plu-CLA also significantly reduced the number of peritoneal metastatic nodules and increased survival in the peritoneal gastric cancer xenograft model. Our results show that intraperitoneal administration of docetaxel-Plu-CLA synergistically inhibits peritoneal metastasis and prolongs survival in a peritoneal gastric cancer model. Therefore, Plu-CLA is a potential intraperitoneal-route carrier for hydrophobic docetaxel for the effective treatment of peritoneal metastatic gastric cancer.

Subclassification of lymphoproliferative disorders in serous effusions: a 10-year experience.

Rare studies have reported the application of multiple ancillary tests to the diagnosis of lymphoproliferative disorder in serous effusions. In the current study, the authors evaluated the effectiveness of using an algorithm for the triage of serous effusions and the contribution of ancillary studies to achieve a specific subtype of lymphoproliferative disorder.

Multiplex sequencing for EZH2, CD79B, and MYD88 mutations using archival cytospin preparations from B‐cell non‐Hodgkin lymphoma aspirates previously tested for MYC rearrangement and IGH/BCL2 translocation

Gene rearrangements and specific translocations define some B‐cell non‐Hodgkin lymphoma (NHL) subtypes. Genome‐wide mutational studies have revealed recurrent point mutations with prognostic implications. The goals of this study were to evaluate the feasibility of applying a multiplex mutation assay to archival cytospin preparations (CPs) and to investigate the rate of EZH2, CD79B, and MYD88 mutations in B‐cell NHL samples previously tested for MYC rearrangement and/or IGH/BCL‐2 translocation.

Cytomorphologic findings of B-cell lymphomas with concurrent IGH/BCL2 and MYC rearrangements (dual-translocation lymphomas)

B‐cell lymphomas with concurrent IGH/BCL2 and MYC gene rearrangements, termed dual‐translocation or double‐hit lymphomas (DTLs), rarely are identified. They usually are characterized by highly aggressive behavior, a poor prognosis, and complex karyotypes. The objective of this study was to review and describe the cytomorphologic findings in different types of cytologic preparations and clinicopathologic characteristics of patients with DTLs.

Epstein-Barr virus encoded RNA detected by in situ hybridization using cytological preparations.

Detection of Epstein–Barr virus (EBV) status might help in the diagnosis of EBV‐related neoplasms. The rate of successful assays for the detection of EBV‐infected cells in cytological preparations has not been fully explored. Our aims were to examine the rate of successful in situ hybridization (ISH) assays for EBV‐encoded RNA (EBER) in cytological specimens and to explore reasons for failure.

Patient-derived tumor xenograft models established from samples obtained by endobronchial ultrasound-guided transbronchial needle aspiration

OBJECTIVES There has been limited utility for laboratory tumor models to predict clinical performance of cancer drugs. Clinical drug trials usually recruit patients that have advanced disease, therefore preclinical tumor models that closely reflect this characteristic will be more reliable to test candidate drugs. We evaluated the use of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) to sample metastatic lymph nodes in patients to establish patient-derived tumorxenograft (PDX) models of advanced lung cancer. MATERIALS AND METHODS Cell suspensions from TBNA aspirates were implanted into the subcutaneous tissue of NSG (NOD scid) gamma) mice. The success rate of PDX establishment was associated with tumor histopathology and the cellularity and cytopathological diagnosis of the primary EBUS-TBNA samples. RESULTS From December 2011 to June 2012, 19 patients were enrolled in this study. Successful engraftment was achieved in 8/19 cases (42.1%). The duration between inoculation and tumor formation averaged 62.4 days (13-144 days). The engrafted tumors included 3 adenocarcinomas (3/12: 25%), 2 squamous cell carcinomas (2/3: 67%), 1 large cell carcinoma (1/1: 100%), and 2 small cell carcinomas (2/3: 67%). CONCLUSION EBUS-TBNA samples can be used for establishment of tumor xenograft model in immunodeficient mice.

Cytomorphological and clinicopathological spectrum of pulmonary marginal zone lymphoma: the utility of immunophenotyping, PCR and FISH studies

To review cytomorphological criteria and clinicopathological findings in combination with ancillary tests for the specific diagnosis of pulmonary marginal zone lymphoma (MZL) in fine needle aspiration (FNA) specimens.

Negative images of crystalline immunoglobulin in crystal storing histiocytosis: A potential cytologic mimic of mycobacteria in smears

Two cases are described of crystal storing histiocytosis (CSH) associated with extranodal marginal zone lymphoma, presenting as lung and subcutaneous masses respectively. Fine‐needle aspiration of subcutis and smears prepared from the resected lung masses showed negative images. Cytology slides of both cases were reviewed to identify cytomorphological features for the differential diagnosis between immunoglobulin crystals and mycobacteria. The crystals in CSH consist of straight and needle shaped rods with pointed or angular edges and are more variable in thickness than the uniformly thin mycobacteria. Mycobacteria show a haphazard distribution, whereas crystals are frequently present in parallel arrays. Small lymphoid or plasma cells are identified in the background of CSH, whereas a necrotic and inflammatory background is seen in mycobacteriosis. Additional samples for culture in the case of mycobacteriosis, or flow cytometry and molecular clonality testing in the case of CSH can provide critical data for a definitive diagnosis. Diagn.Cytopathol. 2012.