From Fine Needle Aspiration Cytology Research Articles

To Get the Most out of the Least: BRAF Molecular Evaluation in Melanoma Metastases on Cell Suspension from Fine Needle Aspiration Cytology Needle Rinses

Introduction: Cytological samples from cutaneous melanoma (CM) metastases may be the only biomaterial available for diagnostic and predictive purpose in the clinical practice. BRAF evaluation from cytological samples actually implies the loss of one or more diagnostic smears, or the execution of one or more passes to obtain dedicated cytological samples. We tested BRAF molecular evaluation in CM metastases on cell suspension obtained from fine needle aspiration cytology (FNAC) needle rinses. Methods: Forty-two patients with lymph node enlargements and a previous CM were enrolled. Patients were submitted to FNAC, and direct smears and cell-block were prepared for diagnostic purpose. The needle was carefully flushed in a vial containing 350 μL of nuclease-free water and a cell suspension was obtained for BRAF molecular evaluation. Molecular evaluation was also performed on histological samples for statistics. Results: The series included 35 CM metastases and 7 reactive lymphadenopathies. Three cases resulted inadequate and adequacy was 92.9%. BRAF V600E/Ec mutations were found in 7 out of 32 (21.9%) CM metastases cases. BRAF mutations other than V600E/Ec were found in 2 out of 32 (6.25%) cases. Sensitivity, specificity, positive predictive value, and negative predictive value resulted 100%. Conclusion: BRAF molecular evaluation in CM metastases on cell suspension obtained from FNAC needle rinses is a time-sparing and accurate technique allowing to spare biomaterial in the clinical setting.

Detection of driver mutations in plasma cell-free nucleic acids in differentiated thyroid neoplasm.

This proof-of-concept paper demonstrates that driver mutations can be detected in plasma in differentiated thyroid tumors, and we were able to detect mutations in upto 80% malignant thyroid nodules. Additionally, cancer subtypes could also be predicted using a 8-gene panel. In almost 90% follicular adenoma, rat sarcoma virus (RAS) mutations were detectable. There was a strong agreement between driver mutations found in plasma samples, FNAC materials, and histopathology samples. This has potential as a noninvasive, preoperative diagnostic tool (particularly of clinical importance in indeterminate nodules) and may help in detection of residual tumor after surgery. Future research is warranted to test the role of this tool to detect tumor recurrence. Ultrasonographic (USG) evaluation and fine-needle aspiration (FNA) are cornerstone for evaluation of thyroid neoplasm. Molecular technique including detection of driver mutation from FNA cytology (FNAC) material is an established modality. In this study, we explored the feasibility of using plasma cell-free nucleic acids to identify known driver mutations in differentiated thyroid neoplasm. Patients presenting with thyroid nodules underwent USG with Thyroid Image Reporting and Data Systems scoring and FNAC (Bethesda classification). All patients in Bethesda 3, 4, 5, 6 underwent surgery and histopathological confirmation. Patients in Bethesda 2 (cosmetic concerns, compressive symptoms) underwent surgery, and rest were presumed benign on the basis of USG, FNAC features, and clinical followup.). Endocrinology clinic. Subjects with thyroid nodule. None. Plasma sample, FNA, and histopathology material were evaluated for driver mutations (8-gene panel comprising BRAF-V600E, RET/PTC3, RET/PTC1, TERT promoter, HRAS, NRAS, KRAS, and PAX8-PPARG). A total of 223 subjects were recruited; of these 154 were benign and 69 had differentiated thyroid cancer. We were able to detect driver mutation from plasma in 55 subjects (79.71%) of all malignant patients, and 11 patients in benign category had RAS mutation (follicular adenoma). Rest of the benign nodules did not have any detectable driver mutations. Plasma might be a viable noninvasive alternative source for detection of driver mutations (8-gene panel) in subjects with differentiated thyroid tumors and may have significant clinical utility.

Programmed Death Ligand-1 Testing in Adenocarcinoma Lung: A Comparative Study of Cell Block versus Biopsy.

Immunotherapy currently stands as a novel treatment option, specifically in cases of advanced non-small cell lung carcinoma (NSCLC). Expression of programmed death ligand-1 (PD-L1) in tumor cells forms the mainstay for the use of anti-PD-L1 monoclonal antibodies in the treatment of NSCLC. The objectives of the study were to assess utility of cell blocks for testing of PD-L1 in adenocarcinoma lung and to compare the expression of PD-L1 in cell blocks and the corresponding biopsy specimens. The current study was a prospective case series that included 20 cases of NSCLC-adenocarcinoma lung. Cases included in the study had biopsies performed from lung masses, along with which cell blocks were prepared from fine needle aspiration cytology (FNAC) samples. Testing for PD-L1 was done using the monoclonal PD-L1 antibody, SP-263 clone on the Ventana Benchmark XT system. PD-L1 expression was assessed only in the tumor cells, and cases with >1% expression, cytoplasmic or membranous, in tumor cells were categorized as positive. PD-L1 expression was identified in the biopsy samples of tumor cells of 20% of cases (n = 4/20). In the corresponding cell blocks, PD-L1 expression was identified in the tumor cells of 15% of cases (n = 3/20). Sensitivity and specificity of cell blocks were 75% and 100%, respectively. Positive and negative predictive values were 100% and 94.12%, respectively. PD-L1 testing has both predictive and prognostic implications. PD-L1 testing in cell block samples is a potential alternative, specifically in cases where biopsy tissue is minimal or unavailable.

A Rare Case of Plexiform Schwannoma of the Little Finger and Its Management: A Case Report

Schwannomas are tumors of the Schwann cells found in the myelin sheath. They cause 5% of all benign soft-tissue cancers, occur equally in males and females, and occur later in life. Since they remain asymptomatic, diagnosing and treating them becomes challenging; current guidelines recommend imaging followed by excision. Here, we present a case of a 19-year-old male who presents in an outpatient setting with a history of painless swelling of the fifth digit for the past four years. Past medical history and physicals are unremarkable. Microscopic findings from fine-needle aspiration cytology (FNAC) confirmed the schwannoma diagnosis, showing loosely arranged spindle cells with elongated nuclei with pointed ends dispersed within the myxoid stroma.

An ensemble deep learning for automatic prediction of papillary thyroid carcinoma using fine needle aspiration cytology

Accurately cytopathological diagnosis of Papillary Thyroid Carcinoma (PTC) is of importance for reducing costs and increasing efficiency of treatments. In this paper, we pursue that goal by introducing artificial intelligence (AI) for automatic classification of malignant PTC cell clusters from Fine Needle Aspiration Cytology (FNAC) processed by ThinPrep. High-resolution cytological images obtained with a 40 × objective lens digital camera attached to an Olympus microscope were segmented into fragments and then divided into training, validation, and testing subsets. Fragments are non-overlapped patches containing only regions-of-interest that cover informative tissue structures for making proper diagnoses. Deep learning CNN models were pre-trained and fine-tuned on large-scale ImageNet domain before they were re-trained on cytology fragments. Moreover, we proposed a method to compute certainty of the patient-level prediction that undoubtedly provides additional evidence for reliability and confidence of the prediction. Results showed that the best classification performance on digital FNAC images achieved using DenseNet161, obtaining a mean accuracy of 0.9556 (p < 0.01), a mean sensitivity of 0.9734, and a mean specificity of 0.9405 on yet-to-be-seen test-set. Ensemble learning findings suggested combinations of AdaBoost classifier with multiple CNN models boosted predictive performances, up to 0.9971 accuracy. Moreover, stain normalization introduced by Reinhard increased the predictive ability, outperforming histogram specification, and Macenko methods. Presented findings demonstrate deep learning can integrate into computer-aided diagnosis systems to support cytopathologists in accurate diagnosis of PTC.

Cellphone enabled point-of-care assessment of breast tumor cytology and molecular HER2 expression from fine-needle aspirates

Management of breast cancer in limited-resource settings is hindered by a lack of low-cost, logistically sustainable approaches toward molecular and cellular diagnostic pathology services that are needed to guide therapy. To address these limitations, we have developed a multimodal cellphone-based platform—the EpiView-D4—that can evaluate both cellular morphology and molecular expression of clinically relevant biomarkers directly from fine-needle aspiration (FNA) of breast tissue specimens within 1 h. The EpiView-D4 is comprised of two components: (1) an immunodiagnostic chip built upon a “non-fouling” polymer brush-coating (the “D4”) which quantifies expression of protein biomarkers directly from crude cell lysates, and (2) a custom cellphone-based optical microscope (“EpiView”) designed for imaging cytology preparations and D4 assay readout. As a proof-of-concept, we used the EpiView-D4 for assessment of human epidermal growth factor receptor-2 (HER2) expression and validated the performance using cancer cell lines, animal models, and human tissue specimens. We found that FNA cytology specimens (prepared in less than 5 min with rapid staining kits) imaged by the EpiView-D4 were adequate for assessment of lesional cellularity and tumor content. We also found our device could reliably distinguish between HER2 expression levels across multiple different cell lines and animal xenografts. In a pilot study with human tissue (n = 19), we were able to accurately categorize HER2-negative and HER2-positve tumors from FNA specimens. Taken together, the EpiView-D4 offers a promising alternative to invasive—and often unavailable—pathology services and may enable the democratization of effective breast cancer management in limited-resource settings.

Necessity of Fine-Needle Aspiration in Probably Benign Sonographic Appearance of Thyroid Nodules.

Introduction:The management of thyroid nodules has been proposed based on US features and information obtained from Fine-Needle Aspiration Cytology (FNAC). In this study, we checked the diagnostic value of ultrasound in comparison with FNAC in probably benign nodules of thyroid. Materials and Methods:Patients with thyroid nodules referred to the Radiology department from 2015 to 2020, were classified into five types based on the American College of Radiology/thyroid imaging reporting and data system (TI-RADS) standards. The patients with TI-RADS III-V were examined by FNA biopsy. Subsequently, the collected data of 535 patients having thyroid nodules with TI-RADS III were statistically analyzed.Results:The mean age of the patients was estimated at 46.57. The analysis of TI-RADS III cases examined by the FNA biopsy revealed that 99.1% of the cases were diagnosed with benign lesions. The mean size of benign and malignant nodules was 27mm and 41mm, respectively. There was no significant correlation between the size of the nodules or patients age, and thyroid malignancy (P-values > 0.05).Conclusion:There was a very low chance of malignancy (0.9%) in thyroid nodules with thyroid imaging classification of TI-RADS III. Furthermore, no meaningful correlation was observed between the size of the nodules and their malignancy. Therefore, the use of FNAC, based on the current guidelines, on thyroids for nodules larger than 2.5 cm might need to be revised.

Diagnostic performance of melanocytic markers for immunocytochemical evaluation of lymph-node melanoma metastases on cytological samples

AimsThe diagnosis of metastatic cutaneous melanoma (CM) on lymph node fine needle aspiration samples may be challenging and usually requires confirmation by immunocytochemistry. However, the cytological material could be too...

CYTO HISTOLOGIC CORRELATION OF A LOW GRADE FIBROMYXOID SARCOMAAT AN UNUSUAL LOCATION – A CASE REPORT

BACKGROUND Low grade fibromyxoid sarcoma (LGFMS) is a rare tumour of the deep soft tissues seen in young adults. The histologic features of LGFMS are well established, however the cytologic findings are scantily defined and the distinction from other benign and malignant myxoid soft tissue tumours is difficult. CASE DETAILS A 29 year old male presented with a superficial swelling in the left areolar region for two months. USG revealed a well-defined 2 by 2 cm hyperechoic lesion extending upto the subcutaneous fat. FNA yielded mucoid material and smears showed spindle cells having round to elongated nuclei with mild anisokaryosis, bland chromatin and wispy cytoplasm in a myxoid background. No significant nuclear pleomorphism or mitoses were noted. A diagnosis of myxoid mesenchymal lesion was made. The excised tumour was well circumscribed with grey white cut surface showing myxoid areas. Microscopically, the tumour displayed moderate cellularity with storiform, intersecting and parallel bundles of spindle cells showing mild nuclear pleomorphism. The myxoid areas showed cells in a haphazard fashion floating in abundant mucoid matrix associated with a capillary network. The mitotic index was low. IHC showed tumor cells positive for vimentin and negative for S-100 &amp; SMA. CONCLUSION LGFMS is a rare low-grade malignancy with a high local recurrence rate and late metastasis. The diagnosis of LGFMS can be difficult to render from fine needle aspiration cytology (FNAC) alone because of morphological overlap with other spindle cell and myxoid lesions. Cytohistological correlation and IHC is helpful in arriving at a correct diagnosis.

Molecular testing approaches in thyroid cancer diagnosis

Thyroid cancer (TC) is the most common endocrine malignancy worldwide, with an annual incidence of around 300,000 cases. In recent decades, the incidence of TC in many countries, including Saudi Arabia, has increased significantly. Genomics has allowed multiple mutations to be examined simultaneously across multiple genes, providing a detailed genomic profile for a given tumor. It is understood from a molecular perspective that different signaling pathways may have genetic abnormalities as the primary factors in thyroid tumorigenesis. While there is still early progress in the usage and standardization of targeted next-generation sequencing (NGS) in TC, a major ongoing study to distinguish malignant from benign thyroid nodules from fine-needle aspiration (FNA) has shown promising results that can prevent unnecessary surgery, based on NGS analyses of mutations and gene fusions associated with most TC. The differential diagnosis and malignancy risk stratification of TC require multidisciplinary skills and experience of both ultrasound and the FNA and molecular analysis. The most common TC mutations are point mutations in the BRAF and the chromosome rearrangements of RET/papillary TCs. The transition mechanisms tend to be linked to specific etiological features, which are very crucial while deciding the treatment protocol. On the other hand, FNA cytology has an intrinsic drawback. The findings of infinite cytology cannot differentiate some types of TC, such as follicular adenomas, thyroid follicular carcinomas, or papillary thyroid follicular. Nevertheless, accumulating evidence indicates that molecular diagnostic approaches can overcome these limitations. In this review, we present an updated summary, which focuses primarily on molecular alterations in the tumorigenesis and biomarker investigations of TC.

Risk factors for hypothyroidism in euthyroid thyroid nodule patients with lymphocytic thyroiditis on fine needle aspiration cytology.

Background/AimsLymphocytic thyroiditis as cytology diagnosis from fine needle aspiration (FNA) is frequently detected in patients with thyroid nodules. However, the clinical outcome for upcoming hypothyroid events has been rarely clarified in euthyroid patients.MethodsWe retrospectively reviewed the data of patient who had lymphocytic thyroitidis on FNA cytology of thyroid nodule from January 2005 to December 2010 at a tertiary referral hospital. In total, 109 patients with follow-up thyroid function tests (TFT) were enrolled. Final outcomes included overt and subclinical hypothyroidism with thyroid stimulating hormone (TSH) levels ≥ 10 mIU/L. Potential parameters predicting clinical hypothyroidism were analyzed by multivariate analysis.ResultsOver the mean follow-up duration of 51.6 months, 14 out of 109 patients (12.8%) developed clinical hypothyroidism that required thyroid hormone replacement. The median onset time to hypothyroidism was 16 months (range, 3 to 88) and ≥ 60% of patients experienced clinical hypothyroidism within 1 year. By multivariate analysis, background thyroiditis (relative risk [RR], 9.78; p = 0.004), thyroid peroxidase antibody positivity (RR, 9.90; p = 0.003), nodule size (RR, 1.24; p < 0.001), and initial TSH (RR, 1.47; p = 0.009) were the independent risk factors for predicting hypothyroidism in euthyroid patients.ConclusionsHypothyroidism frequently occurs during the follow-up in euthyroid patients with thyroid nodules which show lymphocytic thyroiditis on FNA cytology. Close surveillance and regular TFT are needed in high-risk patients for upcoming clinical hypothyroidism.

Adequacy of Fine-Needle Aspiration Cytology for Breast Lesions: The SurePath® Liquid-Based Technique versus Conventional Smears

Objective: We aimed to assess the cytological findings from fine-needle aspiration (FNA) of breast lesions with conventional smear (CS) and SurePath® (SP) cytology. Study Design: This was a cross-sectional study of women who underwent FNA from January 2012 to June 2013 for breast lesions with benign ultrasonography impressions in Fortaleza, Brazil. Two groups were formed. The first was composed of 102 samples subjected to CS and the second of 65 samples subjected to SP. The number of smears, the cellularity and the diagnostic hypotheses were compared. Fisher's exact tests with 95% confidence intervals were applied. Results: The women ranged in age from 22 to 75 years. SP cytology indicated greater cellularity than CS for the cystic lesions (p < 0.05). In the CS group, 72.5% of the samples required 3-4 slides, but in the SP group, only 9.2% required a second slide. The cellularity of the cystic samples was significantly greater with the SP method (p < 0.01). Conclusion: In FNA cytology of the breast, SP is a tool that is comparable to CS, but with the added benefits of better cellularity results for cystic lesions and requiring fewer slides for analysis.

Morphometric Study of Nuclei in FNAC of Breast Lesion and its Role in Diagnosis of Malignancy

Introduction: Cytological features obtained from fine needle aspiration cytology(FNAC) are very essential for the preoperative diagnosis in breast carcinoma. The quantitative evaluation of nuclear size and shape using FNAC material can be utilized as a diagnostic tool in breast tumor. The main objectives of our study were (a) to evaluate major axis (MAJX), minor axis (MINX), nuclear area (NA), nuclear perimeter (NP) and nuclear aspect ratio (NAR) using morphometric techniques, (b) to compare these nuclear parameters with their variability in benign and malignant cases and evaluate suitable cut off values and (c) to study the correlation of these parameters with cytological grades. Materials and Methods: Nuclear parameters were assessed in 50 cases of invasive ductal carcinoma and 50 cases of benign breast tumors by image morphometric technique on FNAC slides and statistical analysis was performed. Results: Mean MAJX, MINX, NA, NP and their variability were significantly greater (p 10.70 micron (specificity=0.98), (b) MINX>7.53 micron (specificity=0.94), (c) NA>60.61 micron2 (specificity=0.98) and (d) NP>27.81 micron (specificity=0.96). Conclusion: Morphometric parameters related to nuclear size and variability evaluated from FNAC material were significantly larger in malignant cases than the benign and they can be gainfully exploited in the diagnosis of breast carcinoma. Again these parameters showed mild positive correlation with the cytological grades.

Thyroglobulin in Washout Fluid From Lymph Node Fine-needle Aspiration Biopsy in Papillary Thyroid Cancer: Large-scale Validation of the Cutoff Value to Determine Malignancy and Evaluation of Discrepant Results

There are still some controversies regarding the cutoff value and the influential factors of thyroglobulin (Tg) concentration in washout fluid from fine-needle aspiration (FNA) biopsy (FNA-Tg) on cervical lymph nodes (LNs) in patients with papillary thyroid cancer (PTC). Our aims were to validate the cutoff value of FNA-Tg in diagnosing malignant LNs on a large scale and to investigate the influential factors that could result in the discrepancy between the final diagnosis and FNA-Tg. We conducted a retrospective cohort study based on hospital records with 528 cases of FNA-Tg measurement from 419 PTC patients. The cutoff value of FNA-Tg was obtained from receiver operating characteristic analysis with final diagnosis. Binary logistic regression analysis was performed to investigate the influential factors. In the final diagnosis, 190 LNs were malignant, and 338 LNs were benign. The median FNA-Tg was 521.2 (3676.8) ng/mL in malignant LNs, and 0.1 (0.2) ng/mL in benign LNs. The optimal cutoff value of FNA-Tg in distinguishing malignant LNs from benign LNs was 1.0 ng/mL (sensitivity, 93.2%; specificity, 95.9%) in all cases. Combining FNA-Tg and FNA cytology showed superior diagnostic power (sensitivity, 98.4%; specificity, 94.4%) when compared with diagnostic strategy using either FNA cytology or FNA-Tg alone. FNA-Tg, serum TSH, and serum Tg were higher in nonthyroidectomized patients than in thyroidectomized patients (P < .001, respectively). FNA-Tg was correlated with serum TSH and Tg levels (P < .001, respectively), and binary logistic regression analysis showed that serum TSH suppression and serum Tg presence independently affected the diagnosis made by FNA-Tg. Our results validated 1.0 ng/mL of FNA-Tg as a cutoff value for diagnosing LN metastasis of PTC and suggested that serum TSH suppression and serum Tg presence should be considered in diagnosing LN malignancy with FNA-Tg in PTC patients.

Chromogenic in situ hybridization to detect EGFR gene copy number in cell blocks from fine-needle aspirates of non small cell lung carcinomas and lung metastases from colo-rectal cancer

BackgroundSeveral studies demonstrated that epidermal growth factor receptor (EGFR) gene copy number (GCN) correlates to the response to tyrosine kinase inhibitors in non small cell lung cancer (NSCLC) and to anti-EGFR monoclonal antibodies (MoAbs) in metastatic colorectal cancer (CRC). In the presence of lung nodules, cytology is often the only possible diagnostic approach. Chromogenic in situ hybridization (CISH) is an alternative technique to fluorescence in situ hybridization (FISH), but its feasibility in detecting EGFR GCN in cell blocks from fine-needle aspiration cytology (FNAC) of lung nodules has not yet been established.MethodsWe evaluated the feasibility of CISH on 33 FNAC from 20 primary NSCLC (5 squamous carcinomas, 8 large cell carcinomas and 7 adenocarcinomas) and 13 lung metastases from CRC.ResultsOf the 33 FNAC analyzed by CISH, 27 (82%) presented a balanced increase in EGFR gene and chromosome 7 number: 10 cases (30%) showed a low polysomy, 15 (45%) a high polysomy and 2 (6%) NSCLC were amplified. No significant differences between NSCLC and CRC lung metastases were found in relation to disomic or polysomic status. In addition, no correlation between EGFR GCN and EGFR immunohistochemical overexpression was found. Furthermore, we compared CISH results with those obtained by FISH on the same samples and we found 97% overall agreement between the two assays (k = 0.78, p < 0.0001). Two cases were amplified with both assays, whereas 1 case of NSCLC was amplified by FISH only. CISH sensitivity was 67%, the specificity and positive predictive value (PPV) was 100%, and the negative predictive value (NPV) was 97%.ConclusionsOur study shows that CISH is a valid method to detect EGFR GCN in cell blocks from FNAC of primary NSCLC or metastatic CRC to the lung.

Low‐grade fibromyxoid sarcoma is difficult to diagnose by fine needle aspiration cytology: a cytomorphological study of eight cases

Low-grade fibromyxoid sarcoma (LGFMS) is an uncommon neoplasm with bland morphology and an indolent clinical course, although metastases may develop in approximately 5-10% of the cases. The diagnosis of LGFMS can be difficult to render from fine needle aspiration cytology (FNAC) alone because of morphological overlap with other spindle cell and myxoid lesions. To determine cytological criteria for LGFMS by reviewing FNAC aspirates in eight cases and to compare the findings with those in subsequent histological sections. FNAC slides were reviewed from eight patients with subsequently excised tumours diagnosed as LGFMS. Of these patients, six also had core needle biopsies (CNB). Cytogenetic and/or molecular analysis was carried on all tumours. The patients were six men and two women ranging in age from 26 to 78 years. Tumours arose in the deep soft tissues of the thigh (n = 5), shoulder girdle (n = 1) or upper arm (n = 1) and one in the subcutaneous tissue of the abdominal wall. Cytological features included clusters of bland spindle and round/polygonal cells embedded in a collagenous and myxoid matrix along with dissociated, uniform or slightly/moderately pleomorphic spindle cells, bare nuclei and fragments of collagen and myxoid tissue in varying proportions. Unequivocal sarcoma was diagnosed in two aspirates, but mitoses were absent in all cases. In three cases, the diagnosis was inconclusive with regard to benignity or malignancy, while three were erroneously diagnosed as benign spindle cell lesions. Although the diagnosis was suggested on three of six CNB, these presented similar diagnostic problems. There were no cytomorphological findings in FNAC to allow for a clear cut separation of LGFMS from other spindle cell or myxoid lesions, but high-grade sarcoma could be excluded. Surgical (incisional or excisional) biopsy or, alternatively, examination of RT-PCR for the FUS/CREB3L or FUS/CREB3L1 fusion transcripts may be necessary to obtain a correct diagnosis.

Comparative validation of c‐kit exon 11 mutation analysis on cytology samples and corresponding surgical resections of gastrointestinal stromal tumours

Studies have shown that c-kit mutation analysis of gastrointestinal stromal tumours (GISTs) obtained by endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) can be routinely performed. We validated c-kit exon 11 mutational analysis on cell block material obtained from fine needle aspiration cytology (FNAC) for diagnostic purposes and compared it with the same analysis in formalin-fixed paraffin-embedded full sections of the corresponding resection specimens. c-kit mutation analysis was done on cell block material obtained from ten cases encountered in our department from 1999 to 2008 on which FNAC was attempted pre-operatively. The findings were compared with analysis on full paraffin section of the corresponding resected tumours in seven cases where patients opted for resection. c-kit exon 11 was examined via bidirectional nucleic acid sequencing. Our results showed 100% concordance for the presence and type of exon 11 mutation in the resected and aspirated tumours in all seven cases. These mutations had diagnostic value when compared with other neoplasms that are part of the cytomorphological differential diagnosis, such as leiomyosarcoma or gastric adenocarcinomas. Molecular cytopathology is a powerful tool that can complement morphology and immunohistochemical assessment of cytological material in routine practice for the diagnosis and prognostication of GISTs. We briefly discuss the advantages and limitations of the fine needle method of obtaining tissue for the diagnosis and prognostication of GISTs, and its current therapeutic strategies.

Identification of carcinoma origin by thyroid transcription factor‐1 immunostaining of fine needle aspirates of metastases

Identification of carcinoma origin by thyroid transcription factor-1 immunostaining of fine needle aspirates of metastases To assess the diagnostic potential of thyroid transcription factor-1 (TTF-1) immunocytochemical staining for identifying the origin of unknown primary cancers from fine needle aspiration cytology (FNAC) of metastases. Immunostaining with TTF-1 was conducted on FNACs suggesting metastatic carcinoma in 128 patients with unknown primary tumours as part of the initial diagnostic procedure. The majority of patients (78) presented with neck masses, four had lung lesions and 46 had masses at other locations. The patients' files were reviewed to obtain data on tumour origin. Thirty-seven of 128 (28.9%) cases stained positive with TTF-1. Pulmonary origin of metastases was confirmed in 27 cases, and four lung lesions were primary lung adenocarcinoma. Two patients had primary and one had mestastatic thyroid carcinomas. In two patients, no follow-up data was available. In one patient, the origin of the primary could not be determined at autopsy. TTF-1 was negative in 18 patients with subsequent diagnosis of metastatic pulmonary cancer and in seven patients diagnosed clinically and radiologically as most probably having a lung carcinoma. In 60 TTF-1 negative patients, the primary tumour originated in other organs (34) or remained unknown (26). TTF-1 immunostaining of FNAC in patients with metastatic carcinoma of unknown origin identified primary pulmonary or thyroid carcinomas in nearly a third of cases. Patients with TTF-1 positive metastases could avoid further extensive diagnostic investigations for the origin of the primary site.

Fine‐needle aspiration for nucleic acid‐ased molecular analyses in breast cancer

With the widespread development of genomic analysis, an accurate and quick method for obtaining high-quality nucleic acids is needed. The objective of the current study was to evaluate the quality and potential use of material obtained from fine-needle aspiration cytology (FNAC). Ultrasound- or palpation-guided FNAC was performed in 124 consecutive patients who had nodular breast lesions. The authors evaluated the amount of messenger RNA (mRNA) obtained, its quality through the RNA Integrity (RIN) score, and the factors that influenced both. For malignant lesions, the authors attempted to correlate estrogen receptor 1 (ESR1) and HER-2 (c-erb-B2) mRNA expression measured by real-time quantitative polymerase chain reaction with estrogen receptor and HER-2 detection obtained by immunohistochemistry (IHC) and/or fluorescent in situ hybridization (FISH) on the surgical specimen. The amount of mRNA obtained was >1 microg in 89.5% of 124 samples (43 benign lesions and 81 adenocarcinomas). Overall, 59.3% of samples yielded >1 microg RNA with a RIN score >6. The most significant predictors of quality and quantity of mRNA were the cytopathologist who sampled the tumors and a diagnosis of cancer versus benign lesion. The median ESR1 expression level, which was expressed as the polymerase chain reaction cycle threshold (CT) level minus the average 18S value (dct), was 17.7 dct in patients with estrogen receptor-negative tumors and 11.1 dct in patients with estrogen receptor-positive tumors. The median HER-2 expression level was 15.1 dct in patients with HER-2-negative tumors and 10.7 dct in patients with HER-2-positive tumors. mRNA expression was concordant with the IHC/FISH evaluation in 90.3% of patients for estrogen receptor status and in 98.5% of patients for HER-2 status. In 70% of cases, FNAC of breast lesions in well trained hands allowed the extraction of mRNA suitable for gene expression analysis.

Risk of Malignancy in Patients With Follicular Neoplasm

To identify clinical and ultrasonographic features that may help in predicting malignant tumors in patients with a diagnosis of follicular neoplasm on findings from fine-needle aspiration cytology (FNAC) because FNAC diagnosis of follicular neoplasm does not differentiate a benign tumor from a malignant tumor. Prospective study of 98 patients having a diagnosis of follicular neoplasm on FNAC. Tertiary cancer referral center. Ninety-eight patients with thyroid nodules diagnosed by FNAC as being a follicular neoplasm. Ultrasonography was performed in each patient, and microcalcifications, echo structure, and echogenicity of the nodules were assessed. All patients underwent surgery. Sensitivity, specificity, positive predictive value, and negative predictive value of ultrasonographic features. Thyroid cancer was diagnosed in 26 patients (27%). Ultrasonographic features (eg, a solid echo structure, microcalcifications, and a hypoechoic pattern) were predictive for malignant neoplasms. The variable associated with the highest sensitivity was the presence of a solid nodule (88.5%), and the variable associated with the highest specificity was the presence of microcalcifications (94.4%). The combination of the 3 ultrasonographic features (solid echo, hypoechoic pattern, and microcalcifications) resulted in a sensitivity of 95.0% and a specificity of 98.6%. Older age, male sex, solitary nodule, and larger nodule size were not predictive for malignant neoplasms in patients with follicular neoplasm cytologic findings. We confirmed that the best compromise between the risk of missing carcinomas and the need for reducing unnecessary surgical procedures would consist of submitting to surgery those nodules presenting a solid echo structure, microcalcifications, or a hypoechoic pattern. Low-risk patients may be observed closely if they are willing to accept a small risk of cancer and if they appreciate the need for a close clinical follow-up.