Fine-Needle Aspiration Cytology In A Case of Chordoma

We report a case of sacrococcygeal chordoma initially diagnosed by fine-needle aspiration cytology. The value of awareness by cytopathologists of the histology of soft tissue tumours is emphasized.     

Pleomorphic adenoma of salivary gland: to what extent does fine needle aspiration cytology reflect histopathological features?

OBJECTIVE: The histological diversity encountered in pleomorphic adenoma may cause diagnostic difficulty in fine needle aspiration (FNA) cytology due to limited and selective sampling. The present study based on 25 histologically confirmed pleomorphic adenoma cases attempts to find out to what extent FNA cytology reflects the histopathological features. METHODS: May-Grunwald-Giemsa and Papanicolaou stained smears, and haematoxylin and eosin stained paraffin sections of 25 pleomorphic adenomas of parotid and submandibular glands were reviewed. The cellularity, which was assessed in a sliding scale of 1+ to 4+, and proportions of epithelial to mesenchymal components in FNA smears and histology was determined and compared. The frequency of morphological features such as squamous metaplastic cells, cells with oncocytic change, acinus formation, mucus globules, papilla formation, giant cells, myxoid and chondroid matrix as well as specific nuclear features was compared between the two diagnostic methods, and the statistical significance was determined using Fisher's exact test of probability. RESULTS: There was complete concordance between cytology and histology with respect to overall cellularity in 14 (56.0%) cases and in the proportions of epithelial to mesenchymal components in 13 (52.0%). Epithelial cells and myxoid matrix were present in all cases. There was no significant difference between smear and tissue section with respect to frequency of squamous metaplasia, oncocytic change, acinus formation, papilla formation, mucus globules, giant cells, nuclear pleomorphism, nuclear chromatin pattern, and mitotic figures. Morphological parameters that were significantly higher in FNAC compared with histology included intranuclear cytoplasmic inclusions (36.0% versus 8.0%, p = 0.0374), nuclear grooves (84.0% versus 48.0%, p = 0.0090), and reniform nuclei (20.0% versus 0.0%, p = 0.0502). Chondroid matrix was the only parameter which was significantly more common in histology than in cytology (44.0% versus 4.0%, p = 0.019). CONCLUSION: FNA cytology demonstrates well most of the histological features of pleomorphic adenoma of salivary gland and may be considered a useful tool in initial assessment of the tumour.     

Leiomyosarcoma of the Breast; Cytological and Histological Features, With A Review of the Literature

This report describes the cytological and histological features of a leiomyosarcoma of the breast. The differential diagnosis of this rare tumour is discussed. Eleven well documented cases of leiomyosarcoma of the breast have been reported previously and details of these are included in this review.     

Borderline Nuclear Abnormality In Cervical Smears: A Cytological Review of 200 Cases With Histological Correlation

The cervical cytology and histology specimens from 200 patients referred to colposcopy with borderline nuclear abnormality were reviewed. Human papillomavirus (HPV)-associated changes were identified in 103 of 200 (53%) referral smears and in 139 of 150 (91%) biopsy specimens. Cervical intraepithelial neoplasia (CIN) was less frequently diagnosed on review compared with the original histopathology reports (30.7%vs 45.4%); the discrepancy was largely attributable to a lower incidence of CINI. There was agreement in the grading of borderline nuclear abnormality in 161 of 200 referral smears following review. Twenty-three smears were upgraded to mild dyskaryosis, whereas 16 were reclassified as negative.     

Determination of Ki67 Growth Fraction and Oestrogen Receptors In Screen-Detected Breast Cancer Using Cytological Preparations

Oestrogen receptor (ER) status of 77 cases of screen-detected breast cancer has been determined using cytological preparations. In 48% ER status was positive, which was the same proportion as that formed in a control group of age-matched patients with symptomatic breast carcinoma. Since the screen-detected group contained more low grade tumours, the percentage of ER-positive cases would be expected to be higher. the reasons for the discrepancy are discussed. Ki67 score has been determined for 41 cases of screen-detected cancer. Ki67 score showed a positive correlation with histological tumour grade and a negative correlation with ER status. However, there was no correlation with tumour size or lymph node status. the Ki67 scores in the screen-detected cancers were essentially similar to those found in an age-matched symptomatic group, but the very low scores were only found in the screened group.     

Comparison of Planimetry and Image Analysis For the Discrimination Between Normal and Abnormal Cells In Cytological Smears of Suspicious Lesions of the Oral Cavity

Light microscope analysis of cytological smears of suspicious lesions of the oral cavity is used as a method for detecting early cancer in the oral cavity. the sensitivity of this approach can be improved by quantitative analysis of the cells in the cytological smears. We have compared the efficiency of planimetry and the Vids V system of image analysis, as quantitative methods for discriminating between normal and abnormal cells in cytological smears of suspicious lesions in the oral cavity. Both methods detected an increase in nuclear area and a decrease in cytoplasmic area in abnormal epithelial cells from dysplastic lesions of increasing severity. However, image analysis was better able to discriminate between benign and malignant cells on the basis of nuclear size. Thus the Vids V system of image analysis is more appropriate than planimetry for quantitative analysis of cytological smears from the oral cavity.     

Dna Ploidy Studies of Benign and Malignant Tumours: Comparison of Flow Cytometry and Image Analysis Techniques Using Two Types of Cytological Specimen

DNA ploidy studies were carried out on Feulgen stained smears and cytocentrifuge preparations from 35 malignant tumours and four benign neoplasms using the CAS image analyser. The smears were prepared from scrapings from fresh tumour tissue whereas the cytocentrifuge preparations were prepared from single nuclear suspensions from paraffin-embedded cell blocks from the same tumour. Histograms obtained by image analysis of the tumour scrapes were compared with those obtained on the cytocentrifuge preparations. Concordant results were obtained in four benign tumours (100%) and 32 malignant tumours (91%). The results obtained by image analysis were also compared with results obtained by flow cytometry of the tumour tissue. Discordant results were obtained for three malignant tumours. Possible reasons for the discrepancy include sampling error, tumour heterogeneity and selective loss of cell populations during processing.     

Cytological investigations on two species of allocreadiid trematodes with special reference to the occurrence of triploidy and parthenogenesis in Allocreadium fasciatusi

Karyotypes and cytological details of gametogenesis were analysed for Allocreadium handiai and A. fasciatusi. Mitotic plates studied in squashes of testes, ovary, eggs and intramolluscan stages showed that A. handiai is a diploid (2n = 14) with six pairs of submetacentrics and one pair of subtelocentrics. Total chromosome length of the diploid complement was 86.57 μm, the largest chromosome measured 8.87 μm (10.24% total chromosome length [TCL]) and the smallest was 2.83 μm (3.28% TCL). Squashes of testes revealed the presence of all stages of spermatogenesis with spermatocytes in various stages of meiotic activity and spermatids containing bundles of spermatozoa. Stages of development of the ovum conclusively proved that reproduction takes place by amphimixis. Mitotic figures of A. fasciatusi, on the other hand, revealed that it is a triploid (3n = 21) with three metacentrics, 12 submetacentrics and six subtelocentrics. The mean total chromosome length of the triploid complement was 137.54 μm. The largest chromosome measured 10.37 μm (7.54% TCL) and the smallest measured 2.67 μm (1.94% TCL). Spermatogenesis was abortive with no evidence of synaptic pairing and spermatozoa were not produced. Eggs remained unfertilized and reproduction was achieved by parthenogenesis which is of mitotic type. The karyotypes of the two species differed not only in the ploidy level but also in the centromeric indexes of certain chromosomes.     

Molecular systematics,GISH and the origin of hybrid taxa in Nicotiana (Solanaceae)

Phylogenetic relationships in the genus Nicotiana were investigated using parsimony analyses of the internal transcribed spacer (ITS) regions of nuclear ribosomal DNA (nrDNA). In addition, origins of some amphidiploid taxa in Nicotiana were investigated using the techniques of genomic in situ hybridization (GISH), and the results of both sets of analyses were used to evaluate previous hypotheses about the origins of these taxa. Phylogenetic analyses of the ITS nrDNA data were performed on the entire genus (66 of 77 naturally occurring species, plus three artificial hybrids), comprising both diploid and polyploid taxa, and on the diploid taxa only (35 species) to examine the effects of amphidiploids on estimates of relationships. All taxa, regardless of ploidy, produced clean, single copies of the ITS region, even though some taxa are hybrids. Results are compared with a published plastid (matK) phylogeny using fewer, but many of the same, taxa. The patterns of relationships in Nicotiana, as seen in both analyses, are largely congruent with each other and previous evolutionary ideas based on morphology and cytology, but some important differences are apparent. None of the currently recognized subgenera of Nicotiana is monophyletic and, although most of the currently recognized sections are coherent, others are clearly polyphyletic. Relying solely upon ITS nrDNA analysis to reveal phylogenetic patterns in a complex genus such as Nicotiana is insufficient, and it is clear that conventional analysis of single data sets, such as ITS, is likely to be misleading in at least some respects about evolutionary history. ITS sequences of natural and well-documented amphidiploids are similar or identical to one of their two parents-usually, but not always, the maternal parent-and are not in any sense themselves 'hybrid'. Knowing how ITS evolves in artificial amphidiploids gives insight into what ITS analysis might reveal about naturally occurring amphidiploids of unknown origin, and it is in this perspective that analysis of ITS sequences is highly informative.     

Fine needle aspiration (FNA) in the diagnosis of soft tissue tumours; a review of 22 years experience

FNA plays an important role in preoperative diagnosis of soft tissue tumours. A close clinical/morphologic cooperation is essential. FNA should be performed on the most accessible part of the tumour, avoiding penetration of the deep portions of the tumour. Needles 0.7 mm (22 G) are recommended. For deep lesions, needles with a stylet should be used. After the FNA, tattooing of the aspiration channel is recommended, and the channel is surgically removed together with the tumour, if a sarcoma. Material from the FNA can be used for additional examinations, i.e. electron microscopy, immunohistochemistry, DNA ploidy analysis and chromosomal analysis. Those techniques are of great importance in the differential diagnosis, particularly in the paediatric small/round cell tumours. the majority of sarcomas can be defined as low grade or high grade malignant in FNA. For malignancy grading the following parameters are used: cellularity, pleomorphism, chromatin pattern, nucleolar structure, mitotic figures and necroses. Cytodiagnostic details of the most common soft tissue tumours and their differential diagnoses are presented.     

Chromosomal Abnormality T(9;22)(Q22;Q12) In an Extraskeletal Myxoid Chondrosarcoma Characterized By Fine Needle Aspiration Cytology, Electron Microscopy, Immunohistochemistry and Dna Flow Cytometry

A multidisciplinary approach was taken to characterize a soft tissue tumour. In smears prepared from aspirated material, uniform tumour cells, embedded in a myxoid matrix and partly arranged in a lace-like pattern, were found. Histopathology showed a lace-like pattern of cells in a matrix of hyaluronidase-stable mucins. Cytoplasmic positivity for S-100 protein was found in some tumour cells. Electron microscopic analysis revealed intracisternal aggregates of microtubules. All these features are consistent with the diagnosis of extraskeletal myxoid chondrosarcoma (EMC). DNA flow cytometry showed a diploid DNA content. Cytogenetic examination revealed the tumour karyotype 45, XY, t(2;11)(q31;p15), t(9;22)(q22.3;q12), dic(13;22)(p11;p13). Because similar 9;22-translocations have been described in two other cases of EMC, we conclude that t(9;22)(q22–31;q11–12) is a specific rearrangement in this tumour type. Cytogenetic analysis may thus be of diagnostic value in the examination of tumours with this and similar histologies.     

Chromosomal abnormalities and DNA image cytometry of haematological neoplasms in fine needle aspirates of lymph nodes

The current diagnostics of haematological neoplasms along with morphological analysis, immunophenotyping and molecular analysis inevitably includes cytogenetic analysis. In this work the possibility of cytomorphological subclassification of haematological neoplasms from lymph node fine needle aspirates was examined without depending upon the referential histological diagnosis and cytogenetic analysis. In addition, the feasibility of cytogenetic analysis of the material obtained by lymph node fine needle aspiration (FNA) was examined. By analysing the findings of cytogenetic analysis and DNA image cytometry, it was decided to examine the possibility of comparing the findings and supplementing diagnostic possibilities of these methods. In 15 cases cytological diagnoses and cytogenetic analysis of haematological neoplasms were performed on the material obtained by lymph node FNA. In 12 of 15 cases histological diagnosis was made separately. A good cytohistological correlation was available in 9 of 12 cases (75%). Cytomorphological diagnoses in 10 of 15 cases (76%) were confirmed by the finding of a specific chromosomal translocation. In two cases cytological diagnosis did not correlate with the histological diagnosis and was confirmed only with specific chromosomal translocations. The lymphocytes obtained by lymph node FNA were adequate material for cytogenetic analysis - in 15 of 18 (83%) cases mitoses in cell cultures were obtained. In 13 of 15 (87%) cases clonal chromosomal abnormalities were detected, whereas in 2 of 15 (13%) cases a normal karyotype was found. DNA image cytometry was performed on nine samples, whereas in six samples the material was not sufficient. Although a small number of samples was analysed in the cases with identical cytomorphological diagnoses, the analysed histograms regarding the DNA index values showed heterogeneity. In conclusion, a cell culture sampled by FNA of lymph nodes is an adequate method for the chromosomal analysis. The specific cytogenetic abnormality associated with cytological diagnosis provides an opportunity to make a definitive diagnosis and provides a powerful approach when reference diagnosis on biopsy material cannot be obtained.     

Detection and Analysis of DNA Damage in Mouse Skeletal Muscle In Situ Using the TUNEL Method

Terminal deoxynucleotidyl transferase (TdT) deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) is the method of using the TdT enzyme to covalently attach a tagged form of dUTP to 3’ ends of double- and single-stranded DNA breaks in cells. It is a reliable and useful method to detect DNA damage and cell death in situ. This video describes dissection, tissue processing, sectioning, and fluorescence-based TUNEL labeling of mouse skeletal muscle. It also describes a method of semi-automated TUNEL signal quantitation. Inherent normal tissue features and tissue processing conditions affect the ability of the TdT enzyme to efficiently label DNA. Tissue processing may also add undesirable autofluorescence that will interfere with TUNEL signal detection. Therefore, it is important to empirically determine tissue processing and TUNEL labeling methods that will yield the optimal signal-to-noise ratio for subsequent quantitation. The fluorescence-based assay described here provides a way to exclude autofluorescent signal by digital channel subtraction. The TUNEL assay, used with appropriate tissue processing techniques and controls, is a relatively fast, reproducible, quantitative method for detecting apoptosis in tissue. It can be used to confirm DNA damage and apoptosis as pathological mechanisms, to identify affected cell types, and to assess the efficacy of therapeutic treatments in vivo.     

A Retrospective Study of Clinicopathologic Outcomes of Nodules With Hürthle Cell Cytology and the Thyroid Nodule App (TNAPP) Ultrasound Recommendations

ObjectivesHürthle cells are a common finding on thyroid fine-needle aspiration, but when they are the predominant cytology, they represent a difficult diagnostic challenge. The Thyroid Nodule App (TNAPP) is a new, publicly available web application utilizing ultrasound (US) features based on the updated 2016 American Association of Clinical Endocrinologists clinical practice guidelines for thyroid nodule management. This pilot study was performed to assess the TNAPP recommendations and surgical pathology outcomes of Hürthle cell-predominant thyroid nodules.MethodsA retrospective review of nodules with Bethesda III (atypia of undetermined significance with Hürthle cells) or Bethesda IV (suspicious for Hürthle cell neoplasm) cytology, for which surgery was performed between 2017 and 2021, was conducted. TNAPP US categories 1, 2, and 3 (low, intermediate, and high risk, respectively) were assigned based on nodule characteristics, and clinical management recommendations were recorded. Results were compared with histology-proven diagnoses.ResultsFifty-nine nodules in 57 patients where surgical pathology was available were analyzed with the TNAPP algorithm. Of the 59 nodules, 4 were US category 1 (low risk/suspicion), 40 were US category 2 (intermediate risk/suspicion), and 15 were US category 3 (high risk/suspicion). All US category 1 nodules were benign, while 30% of the US category 2 and 40% of the US category 3 nodules were malignant. Of the patients who had molecular marker testing with ThyroSeq, 22 out of 29 (76%) were positive, indicating either an intermediate or high risk of malignancy, 7 of which were malignant.ConclusionThis preliminary study suggests that TNAPP is a useful clinical tool for sonographic assessment of thyroid nodules with Hürthle cell cytology.     

Management of Sonographically Suspicious Thyroid Nodules 1 cm or Smaller and Candidacy for Active Surveillance: Experience of a Tertiary Center in China

ObjectiveOur objective was to investigate the management of patients with asymptomatic suspicious thyroid nodules ≤1 cm.MethodsWe retrospectively reviewed medical records of patients with sonographically suspicious thyroid nodules ≤1 cm and without distant metastases, suspicious lymph node metastasis (LNM), or extrathyroidal extension (ETE).ResultsOf the 386 enrolled patients, 174 (45.1%) had immediate surgery (IS), while 212 (54.9%) underwent active surveillance (AS). In the IS group, 166 (95.4%) patients were confirmed as having papillary thyroid microcarcinoma. LNM and ETE were observed in 24.7% and 2.4% cases, respectively. In the AS group, nodule size increased by ≥3 mm in 11 (5.2%) patients and 39 (18.4%) had a >50% increase in nodule volume after a median follow-up of 12 months. Nodules with smaller volume at diagnosis were more likely to increase in volume later. Newly suspicious LNM was detected in 23 (10.8%) patients. Delayed surgery (DS) was performed in 101 patients, with 27 showing disease progression. ETE and LNM were detected in 3% and 36%, respectively, of patients with papillary thyroid microcarcinoma. Compared with IS, tumors in the DS group more frequently showed lateral LNM and capsular invasion (P < .05). No patient had recurrence or died of thyroid cancer during postoperative follow-up (median 26 [4-60] months).ConclusionsIS or DS of patients with asymptomatic suspicious thyroid nodules ≤1 cm was relatively high in China. The inertia of low-risk nodules and the effectiveness of DS for those that progressed make AS a feasible strategy.     

Diagnostic Role of Cell-free DNA Integrity in Thyroid Cancer Particularly for Bethesda IV Cytology

BackgroundThe cell-free DNA integrity index (cfDI) is promising for the differentiation between malignant and benign tumors, but little data has been reported on thyroid cancer (TC). We explored its diagnostic role in TC, mainly in cases of Bethesda category IV.MethodscfDI was evaluated by quantitative real-time polymerase chain reaction using 2 primer sets to identify cell-free DNAs (cfDNAs) Alu83 and Alu244. Blood samples were collected from 85 patients with thyroid nodules (18 papillary [PTC], 21 follicular [FTC], 21 medullary, and 25 benign thyroid nodules [BTN]) before fine-needle aspiration cytology and surgical treatment and also from 25 patients with autoimmune thyroid disease (ATD) and 25 healthy subjects (HS).ResultscfDNA Alu244 concentration ≥6.95 ng/mL and cfDI ≥0.3 were excellent sensitive and specific tests to discriminate TC particularly cytologically indeterminate thyroid nodules (Bethesda IV) from the control groups (BTN, ATD, and HS). The levels of both cfDNA Alu83 and Alu244 were decreased while cfDI was increased significantly in medullary compared with FTC and PTC, with a nonsignificant difference between the latter subgroups. There was a significantly positive correlation between both cfDNA Alu83 and Alu244 with the T-classification of TNM staging and capsular invasion among PTC and FTC patients and between cfDI with Bethesda categories. Additionally, ATD had significantly higher cfDNA Alu83 and lower cfDI than HS.ConclusioncfDI is a useful noninvasive molecular biomarker in TC that correlates with the Bethesda classification and histopathology. Tumor size and capsular invasion were correlated with quantitative cfDNA among PTC and FTC.