Immacolata Cozzolino

Cytologic, flow cytometry, and molecular assessment of lymphoid infiltrate in fine-needle cytology samples of Hashimoto thyroiditis

The thyroidal lymphoid infiltrate (TLI) in Hashimoto thyroiditis (HT) represents the substrate from which thyroid lymphoma may arise. The objective of the current study was to classify the TLI in HT by comparing the cytologic features with flow cytometry (FC) data and evaluating the κ/λ light chain ratio and its molecular assessment.

Cyclin D1 and D3 overexpression predicts malignant behavior in thyroid fine-needle aspirates suspicious for Hurthle cell neoplasms

Thyroid fine‐needle aspiration (FNA) samples that feature a follicular‐patterned, monotonous Hurthle (oncocytic) cell population cannot be diagnosed reliably. The authors of this report recently identified cyclin D3 overexpression on histologic sections of Hurthle cell carcinoma. In this study, they assessed the diagnostic value of cyclin D3 immunohistochemistry added to routine cytology.

Impact of endoscopic ultrasound‐guided fine needle aspiration (EUS‐FNA) in lymph nodal and mediastinal lesions: A multicenter experience

Endoscopic ultrasound‐guided fine needle aspiration (EUS‐FNA) is an established procedure in lung cancer (LC) staging and in the diagnosis of mediastinal masses. Most of the experiences reported refer to single specialized centers where dedicated teams of endoscopists and pathologists perform the procedure. We report the EUS‐FNA experience of a cooperation group involving clinicians and cytopathologists from three hospitals. Fifty‐seven consecutive EUS‐FNA of mediastinal nodes in LC patients, eight mediastinal and two subdiaphragmatic masses were collected in 3 years. EUS‐FNA was performed by two endoscopists and three experienced pathologists. On‐site evaluation was performed in all cases by the three cytopathologists. Lymph node negative cases underwent surgery, which confirmed the cytological diagnoses but also detected two false negatives. Four of the 10 EUS cytological diagnoses of mediastinal and subdiaphragmatic masses were histologically confirmed. All EUS diagnoses were blindly reviewed by three pathologists to assess intra and interpersonal reproducibility. FNA‐EUS diagnoses were: 10 inadequate (17%), 10 negative (17%), 4 suspicious (7%) and 33 positive (59%). Diagnoses of mediastinal and subdiaphragmatic masses were: relapse of lung carcinoma (3), mesenchimal tumor not otherwise specifiable (3), gastrointestinal stromal tumor (GIST) (1), esophageal carcinoma (2) and paraganglioma (1). The sensitivity attained was 85% and the specificity 100%; revision of the slides demonstrated a significant diagnostic reproducibility of the three cytopathologists (P < 0.5). The sensitivity and specificity attained were similar to those reported in the literature suggesting that experienced cytopathologists and endoscopists from different institutions can employ the same procedure reaching comparable results. Diagn. Cytopathol. 2010.

Lymph node fine needle Cytology in the staging and follow-up of Cutaneous Lymphomas

BackgroundLymph nodal involvement is an important clinical-pathological sign in primary cutaneous lymphoma (PCL), as it marks the transformation/evolution of the disease from localized to systemic; therefore the surveillance of lymph nodes is important in the staging and follow up of PCL. Fine needle cytology (FNC) is widely used in the diagnosis of lymphadenopathies but has rarely been reported in PCL staging and follow-up. In this study an experience on reactive and neoplastic lymphadenopathies arisen in PCL and investigated by FNC, combined to ancillary techniques, is reported.MethodsTwenty-one lymph node FNC from as many PCL patients were retrieved; 17 patients had mycosis fungoides (MF) and 4 a primary cutaneous B-cell lymphoma (PBL). In all cases, rapid on site evaluation (ROSE) was performed and additional passes were used to perform flow cytometry (FC), immunocytochemistry (ICC) and/or polymerase chain reaction (PCR) to assess or rule out a possible clonality of the corresponding cell populations.ResultsFNC combined with FC, ICC, and PCR identified 12 cases of reactive, non specific, hyperplasia (BRH), 4 dermatopathic lymphadenopathy (DL), 4 lymph nodal involvement by MF and 1 lymph nodal involvement by cutaneous B-cell lymphoma.ConclusionsFNC coupled with ancillary techniques is an effective tool to evaluate lymph node status in PCL patients, provided that ROSE and a rational usage of ancillary techniques is performed according to the clinical context and the available material. The method can be reasonably used as first line procedure in PCL staging and follow up, avoiding expensive and often ill tolerated biopsies when not strictly needed.

Immunoglobulin heavy-chain fluorescence in situ hybridization-chromogenic in situ hybridization DNA probe split signal in the clonality assessment of lymphoproliferative processes on cytological samples.

The human immunoglobulin heavy‐chain (IGH) locus at chromosome 14q32 is frequently involved in different translocations of non‐Hodgkin lymphoma (NHL), and the detection of any breakage involving the IGH locus should identify a B‐cell NHL. The split‐signal IGH fluorescence in situ hybridization‐chromogenic in situ hybridization (FISH‐CISH) DNA probe is a mixture of 2 fluorochrome‐labeled DNAs: a green one that binds the telomeric segment and a red one that binds the centromeric segment, both on the IGH breakpoint. In the current study, the authors tested the capability of the IGH FISH‐CISH DNA probe to detect IGH translocations and diagnose B‐cell lymphoproliferative processes on cytological samples.

Cytological and molecular features of papillary thyroid carcinoma with prominent hobnail features: a case report.

Background: Papillary thyroid carcinoma (PTC) with prominent hobnail features is a recently recognized PTC variant. Its histological hallmark is represented by elongated cells showing a high nuclear/cytoplasmic ratio and a hobnail appearance. Few histological studies have been performed showing aggressive clinical and pathological features. Thus, a better patient management might benefit from its early diagnosis on fine needle aspiration (FNA) samples. To date, the FNA cytology of PTC with prominent hobnail features has not been described. Case Report: We retrospectively analyzed the FNA taken from a histologically proven PTC with prominent hobnail features. Although there was a certain degree of morphological overlap with the tall cell variant, some peculiar cytological features were observed. In particular, the novel ‘comet-like’ cell feature could represent the cytological counterpart of the histological hobnailing. V600E BRAF mutation was observed on a matched cytohistological specimen. Conclusion: Further investigation is required to validate our diagnostic criteria, as the recognition of PTC with prominent hobnail features on FNA may prospectively enable its preoperative diagnosis, suggesting more aggressive neck surgery.

Lymph nodal Merkel cell carcinoma: primary tumor or metastasis from unknown primary site?

To the Editor, Merkel cell carcinoma represents rare, aggressive skin cancer with neuroendocrine differentiation.1– 4 Most Merkel cell carcinomas develop within the skin of the head and neck or on the extremities; in <10% of cases, the trunk or mucus membranes are involved.1– 5 Regional lymph node metastases occur in 50–66% of cases. In 1992, Eusebi et al.5 described eight cases of nodal Merkel cell carcinoma in the absence of an identifiable primary malignancy. Since that time, other authors3,4 have reported cases of ‘primary’ nodal Merkel cell carcinoma and have raised the issue of whether this phenomenon truly occurs or whether such nodal involvement represents, instead, metastasis from an occult tumor.3,4 In such cases, strict criteria should be applied before rendering a diagnosis of primary nodal Merkel cell carcinoma.2 In addition, there is need for specific immunohistochemical evidence to support the diagnosis.1– 5 Herein, we report a case of recurring Merkel cell carcinoma in two distinct lymph node sites with no evidence of an associated primary carcinoma. A 77-year-old male underwent excision of an inguinal node of 35 mm in greatest dimension located on his right side. Histopathologically, the node was replaced by an undifferentiated malignancy. The tumor exhibited a solid configuration and was composed of small cells with monomorphic, ovoid vesicular nuclei and scant cytoplasm (Fig. 1A); mitotic figures were numerous. Immunoperoxidase staining revealed positivity for cytokeratin AE1/AE3, NSE, chromogranin and CK20 with paranuclear dots (Fig. 1B). There was a lack of expression of LCA, CK7 and TdT. As all clinical and imaging investigations, including a PET–CT, were Fig. 1. A) The node is effaced by a solid tumor with cells that are monomorphous, small and ovoid with vesicular nuclei and scant cytoplasm (hematoxylin–eosin ×270). B) Immunocytochemical positivity for CK20 with paranuclear dots (ABC immunostain ×270). C) Needle aspiration smear showing a dispersed small cell population with scant clear cytoplasm; the nuclei are roundpolygonal with fine chromatin and inconspicuous nucleoli (Diff-Quik ×270). D) Immunocytochemistry on cell block showing CK20 positivity with a paranuclear dot-like pattern (ABC immunostain ×270).

Cytological and molecular diagnosis of solid variant of papillary thyroid carcinoma: A case report

Papillary thyroid carcinoma (PTC) composed by predominant solid areas is diagnosed as a distinct variant on histological samples. Here we present a case of PTC recognized preoperatively by fine needle cytology as a solid variant. This diagnosis was made by combining cytology with the detection of the BRAFVK600-1E mutation, the molecular hallmark of the solid variant of PTC. Histological and molecular evaluation of the surgical specimen confirmed this pre-operative diagnosis. Thus combining cytology to BRAF molecular analysis is useful to refine the cytological diagnosis of this variant also on FNC specimens.

Cytological and histological detection of amyloid deposits in bone marrow of patients affected by multiple myeloma

Abstract We recently published a study aiming to verify the frequency of amyloid deposits in the bone marrow of patients with multiple myeloma (MM) who did not present any signs or symptoms of systemic amyloidosis, applying the Congo red technique on bone marrow smears obtained by aspiration from the posterior iliac spine. The results suggested that nearly 40% of patients affected by MM may have amyloid deposits in their bone marrow. Subsequently, this finding has not been confirmed by another study performed with histological specimens of bone marrow in a similar clinical setting. To explain this discrepancy, we performed a comparative study on the bone marrows of 36 patients affected by MM, evaluated by both cytological and histological techniques. The results of this study confirm the high frequency of amyloid deposits in the bone marrow of patients affected by MM when the analysis is made on cytological smears, and indicate that the presence of amyloid on marrow smears is confirmed by core biopsies simultaneously performed in only 25% of cases. Should further studies confirm our findings, cytological assessment could be considered a sensitive technique to detect bone marrow amyloid deposits.

Metastasis of colon cancer to the thyroid gland: A case diagnosed on fine-needle aspirate by a combined cytological, immunocytochemical, and molecular approach

Fine‐needle aspiration (FNA) with cytological evaluation reliably diagnoses primary and secondary thyroid neoplasms. However, identifying the primary origin of a metastatic process involving the thyroid gland is challenging. In particular, metastasis of colon cancer to the thyroid gland is very rare. In this case report, a right lobe solid thyroid nodule in a 66‐year‐old male was aspirated. FNA cytology showed necrosis and atypical tall columnar cells; since, the patient at age 60 had undergone surgery for a sigmoid‐rectal cancer metastasizing to the liver and subsequently to the lung, a suspicion of metastasis from colon cancer was raised. This was corroborated by cell‐block immunocytochemistry showing a cytokeratin (CK) 7 negative/CK20‐positive staining pattern; thyreoglobulin and TTF‐1 were both negative. Since KRAS codon 12/13 mutations frequently occur in colon cancer, whereas they are extremely uncommon in primary thyroid tumors, DNA was extracted from the aspirated cells, and KRAS mutational analysis was carried out. The codon 12 G12D mutation was found; the same mutation was evident in the primary cancer of the colon and in its liver and lung metastasis. Thus, a combined cytological, immunocytochemical and molecular approach unquestionably correlated metastatic adenocarcinoma cells aspirated from the thyroid to a colo‐rectal origin. Diagn. Cytopathol. 2010;38:932–935.