Cytological Material Research Articles

From sampling to cellblock: The fully automated journey of cytological specimens.

In recent years, technological innovation have emerged to standardize pathology laboratory processes and reduce the handling of diagnostic samples. Among them is an automatic tissue embedding system that eliminates the need for manual activity in tissue paraffin embedding, thereby improving sample preservation. Unfortunately, this system cannot be used for cytological specimens due to the lack of an effective holder to support the procedure steps. In this study, we evaluated the performance of a commercial polymer matrix to enable and standardize the automatic paraffin embedding of cytological material from different organs and sources. Cytological samples from 40 patients were collected on the matrices and submitted for fully automatic workflow preparation, from formalin fixation until paraffin block, using the Sakura embedding system. Our results demonstrated the feasibility of the automated procedure, from loading cytological sample onto the matrix to obtaining the paraffin cellblock, thereby avoiding manual manipulation of cellular material. All samples resulted adequately processed and paraffin-embedded showing satisfactory tissue permeation by processing reagents, optimal preservation of cytoplasmic and nuclear details, and good quality of staining results on paraffin sections. Automated embedding of cytological samples eliminates the risk of lost specimens, reduces laboratory burden, standardizes procedures, increases diagnostic yield, and ultimately improves patients' management.

Re-Categorization and Risk of Malignancy of Atypical Effusions According to the International System for Reporting Serous Fluid Cytopathology (TIS): An Institutional Experience.

Serous effusion fluids serve as crucial cytological materials in diagnosing the underlying causes of fluid accumulation in various body cavities. The International System for Reporting Serous Fluid Cytopathology (TIS) outlined atypia of undetermined significance (AUS) and suspicious for malignancy (SFM) as two intermediate categories for atypical fluids. In our study, cases diagnosed as "atypical" by an experienced cytopathologist in the Hacettepe University Department of Pathology, between 2014 and 2023, were re-categorized according to TIS. The risk of malignancy (ROM) for AUS and SFM categories was calculated. Additionally, cases were re-evaluated according to the 5-tier categorical system by two observers with different level of experience. A total of 3501 effusion fluids were included in the study, evaluated between 2014 and 2023. 50.7% of the cases were male, and 49.3% were female. 55.2% of the cases were pleural fluid, 41.5% were peritoneal fluid, and 3.3% were pericardial fluid. Two hundred sixty five cases (7.6%) were non-diagnostic, 2160 (61.7%) were negative for malignancy, 111 (3.2%) were AUS, 83 (2.4%) were SFM, and 883 (25.2%) were malignant (M). ROM calculated 27% for AUS, 49% for SFM 49%. The interobserver agreement for AUS was 46% with kappa value of 0.162, and 42% for SFM with kappa value of 0.188. The differentiation of atypical cases into AUS and SFM is effective in determining the ROM. The interobserver agreement tends to be lower in intermediate categories compared to others, emphasizing the potential significance of clinical expertise in diagnosis.

7817 A Rare Case of Intrathyroidal Thyroglossal Duct Cyst Presenting as a Rapidly Expanding Left Thyroid Goiter

Abstract Disclosure: C.M. Godar: None. A.J. Spiro: None. K.F. Brown: None. M.K. Shakir: None. T.D. Hoang: None. Introduction: Thyroglossal duct cysts are the most common congenital cyst in the head and neck, found during childhood as a midline neck mass. We present a rare case of a large intrathyroidal thyroglossal duct cyst with left lateral extension in an adult patient. Case Report: A 50-year-old man with family history of familial adenomatous polyposis presented for thyroid nodule evaluation. Thyroid ultrasound for screening of thyroid cancer showed a 4.8 x 4.5 x 2.5cm mixed cystic/solid hyperechoic thyroid nodule with smooth margins in the left lower thyroid lobe. He reported no compressive symptoms. FNA with standard 27G needle was insufficient for evaluation with Bethesda I result. Attempted syringe aspiration did not produce cystic fluid for analysis. The patient returned to clinic 8 weeks later due to visible enlargement of his anterior neck and new neck pressure with swallowing. Physical exam was significant for a new, firm palpable mass over the left thyroid isthmus. Bedside ultrasound noted expansion of the previously visualized mass now extending into the thyroid isthmus. FNA was reattempted with a 25G needle, and cytologic material was again inadequate. He elected to undergo left hemithyroidectomy. Histologic sections demonstrated a 3.4cm simple squamous, non-keratinizing cyst lining overlying a thickened fibrotic wall with mixed acute and chronic inflammation surrounded by benign thyroid tissue, consistent with an intrathyroidal thyroglossal duct cyst. Discussion: The thyroglossal duct derives from epithelial tissue during embryogenesis and atrophies during weeks 8-10 of gestation. A persistent duct may lead to cyst formation in the presence of remnant epithelial tissue secretions and local neck inflammation. A thyroglossal duct cyst is usually located midline in the anterior neck. This stands in comparison to a branchial cleft cyst, which arises due to failure of atrophy of the second branchial groove during embryogenesis and forms a cyst that is usually located in the upper lateral neck compartment. Other common neck masses include lipomas, dermoid cysts, or ectopic thyroid tissue. This case highlights the rarity of intrathyroidal thyroglossal duct cyst in an adult patient and the importance of definitive diagnosis and appropriate management. Presentation: 6/2/2024

Cytomorphologic analysis of pulmonary neuroendocrine tumors – The physical effect of abrasion and aspiration on cytomorphology

Neuroendocrine tumors of the lung display characteristic cytomorphologic features allowing direct diagnosis. The specificity of these features in distinguishing subtypes of neuroendocrine tumors, and their differences among types of cytologic specimen poses as interpretative potential pitfalls. This study reviewed and compared bronchial, effusion fluid and fine-needle aspiration cytology specimens of neuroendocrine tumors of the lung to address these issues. Histology-proven cytology specimens of neuroendocrine tumors were reviewed for cytomorphological parameters focusing on reported specific neuroendocrine nuclear and background features. Totally, 46 cases (26 bronchial, 11 effusion and 9 aspirate specimens), corresponding to 37 small cell carcinomas, 7 neuroendocrine carcinomas and 2 carcinoids were reviewed. Nuclear moulding (n = 35/37, 95 %), naked nuclei (n = 33/37, 89 %) and marked nuclear irregularity (n = 32/37, 86 %) were the three most common features of small cell carcinoma. The only specific feature for small cell carcinoma was the lack of prominent nucleoli (p = 0.004). For pulmonary carcinoids, in addition to the above features, other features associated with neuroendocrine carcinoma reviewed including crush artifact and necrotic material were absent. Compared to bronchial and aspiration cytology, crush artifact (p < 0.001) and necrotic material (p = 0.014) were absent on effusion fluid specimens and naked nuclei were less frequently seen (p = 0.022), while prominent nucleoli were more often observed (p = 0.005). Nuclear moulding, irregularity and naked nuclei are common but not unique features to small cell carcinomas. Effusion fluid specimens have “cleaner” backgrounds while displaying greater nuclear atypia. The type of cytologic preparation/specimen is an important factor which must be considered during diagnostic interpretation.

Application and Reliability of the Milan System for Reporting Salivary Gland Cytopathology (MSRSGC): An Institutional Study

Introduction: Milan System for Reporting Salivary Gland Cytopathology (MSRGC) was introduced in the year 2015 to facilitate a standardised reporting format and to improve the overall effectiveness of fine-needle aspiration cytology (FNAC) in the preoperative evaluation of salivary gland lesions. Risk of malignancy (ROM) provided in each category guides the physician for appropriate clinical management. This study was performed prospectively for one year to evaluate the diagnostic utility and validate MSRSGC. Materials and Methods: The study included lesions in the salivary gland area, including intraparotid lymph node. Cytology material from both direct FNAC, which had rapid onsite evaluation (ROSE) for adequacy and ultrasonography-guided procedures, was included for analysis. Relevant clinical information and imaging findings were obtained. Slides were reviewed, and one of the six categories was applied using the Milan system. ROM and risk of neoplasm (RON) were calculated for each category. Results: The study included 62 cases of salivary gland FNAC of which 35.5% of the cases had surgical resection specimens. Category IVA formed the major group, and only 1.6% was given category III. The ROM and RON were calculated for each category and were statistically significant and compared with other studies. Conclusion: MSRSGC is the recent classification system, and it suggests less than 10% of cases under category III to avoid it’s over utilisation. MSRSGC is a reliable classification in our population, and it was best with category IVA lesions in our study.

The use of vacuum fine needle aspiration biopsy of small formations in the diagnosis of thyroid cancer

Introduction. Diagnosis and treatment of lesional pathology of the thyroid are important problems in Russia and worldwide. Currently, fine-needle aspiration (FNA) is the main technique of differential diagnosis of benign and malignant tumors of the thyroid. In 2023, clinical guidelines of the European Thyroid Association (ETA) on thyroid lesion treatment were published noting possible minimally invasive treatment of benign and malignant tumors of the thyroid requiring repeat cytologic verification. Despite the proven effectiveness of FNA, its informativity and specificity vary between 60 and 100 %. Additionally, this method is associated with high risk of false negative and false positive results. Therefore, there is a need to improve FNA and minimize its disadvantages.Aim. To evaluate the capabilities of vacuum FNA of small lesions using multiparameter ultrasound in diagnosis of thyroid cancer.Materials and methods. At the scientific and practical problem laboratory Diagnostic Studies and Minimally Invasive Technologies of the Smolensk State Medical University, a device for vacuum FNA was developed. Between 2023 and 2024, patients with thyroid lesions of size &lt;10 mm underwent 25 vacuum FNAs. Correlation analysis with 51 FNAs performed between 2020 and 2023 was performed.Results. A decrease in the number of nondiagnostic (category I per the Bethesda system, 2017) and undetermined (category III per the Bethesda system, 2017) results was observed. Comparison of preoperative cytologic conclusions (category V per the Bethesda system, 2017) with the results of postoperative histological examination in patients of the control group showed benign lesions in 2 (50 %) cases which correspond to false negative results. Vacuum FNA did not show this discrepancy. These results demonstrate a decrease in the probability of false negative results and effectiveness of vacuum FNA.Conclusion. Use of vacuum FNA expedites ultrasound-guided biopsy for small thyroid lesions. Due to high negative pressure (up to –0.8 bar), single-step retrieval of cytologic material is performed which leads to a decrease in the number of nondiagnostic and undetermined cytologic conclusions by 11.7 and 1.9 %, respectively.

"Immunocytochemistry in Cytology: Myth or Reality": Unraveling the Myth - Immunocytochemistry Applications in Thyroid Lesions.

Fine-needle aspiration cytology serves as an important preoperative diagnostic tool for thyroid nodules. Despite its excellent diagnostic accuracy, diagnoses based solely on morphological observation can be challenging. Therefore, various ancillary diagnostic techniques have been applied, including immunocytochemistry (ICC). This review discusses the application and evaluation of ICC in thyroid fine needle aspiration. Currently, three immunostaining preparation methods are available for cytological materials: liquid-based cytology, cell block, and cell transfer. ICC proves valuable in scenarios such as tumour diagnosis, assessment of differentiation and grading of carcinomas, estimation of primary organs in metastatic carcinomas, and detection of gene abnormalities. However, ICC, while useful, is not as accurate as immunohistochemistry and is more difficult to evaluate. If the pitfalls and limitations are understood and effectively navigated, ICC could play a significant role in decreasing the non-diagnostic rate, thus leading to more accurate and valuable diagnoses and reductions in the re-aspiration rate.

Diagnosis of metastatic melanoma by fine-needle aspiration biopsy and primary melanoma by using the skin and mucousa imprints

BACKGROUND: Determination of the cytogenetic origin and morphological type of metastatic or primary tumor determines the prescription of therapy and affects the efficiency of patient’s treatment. Currently, a number of publications evaluate the potential of cytological diagnostics of melanoma. AIM: To evaluate the significance of diagnosing metastatic melanoma by fine-needle aspiration biopsy and primary melanoma by using the skin and mucosa imprints. METHODS: In a retrospective study, a comparative analysis of cytological melanoma samples, in comparison with clinical and anamnestic information and the results of histological, immunohistochemical, and molecular genetic studies, was carried out. Information about 109 patients from the cancer registry of the Altai Regional Oncology Dispensary (Barnaul, Russia) for 2022 was used in the study. Traditional and liquid-based methods for preparing samples were used. The samples were stained using Pappenheim and Papanicolaou methods. In some observations, cytological material was used for molecular genetic studies. Using the cancer registry data of the dispensary, the results of histological and of molecular genetic studies, and a final conclusion was given about each patient. RESULTS: Fine-needle aspiration biopsy was carried out in 80 patients. Tumor smears were obtained from 29 patients. The cytological diagnose “melanoma” was consistent with the data of histological and immunohistochemical studies (p 0.001) for all 109 patients. Melanoma was diagnosed for the first time in 66 (60.5%) patients. In other cases, the process of progression was noted within the period from one year to 20 years. Epidermal melanoma was noted in 101 (92.7%) cases, including 9 patients with acral melanoma, and 2 cases with localization on the vulva. Melanomas of mucosa were found in 5 cases (4.5%): in the rectum and anal canal, vagina, and in 2 cases on the hard palate. Non-epidermal (uveal) melanomas metastases were diagnosed in liver by fine-needle aspiration biopsy in 3 patients (2.8%). Based on the cellular composition, epithelioid cell melanoma was determined in 81 (74.3%) patients, mixed cell in 14 (12.8%) cases, spindle cell in 9 (8.2%), pigmentless in 2 (1.8%) cases and nevoid melanoma in 1 (0.9%) case. The mutation status was determined in 96 patients (88.1%). Of these, in 8 patients it was determined using cytological material. In epidermal melanomas, mutations in codon 600 of exon 15 of the BRAF gene were found in 43 (44.8%) patients, including 2 cases of acral melanoma. Mutations V600K, V600E/Ec were found in one patient each. In mucousal melanomas: Q61R mutation was found in exon 3 of the NRAS gene (vaginal melanoma), G12C mutation was identified in exon 2 of the NRAS gene (anal canal melanoma). In uveal melanomas, the assessed mutations were absent (it is necessary to determine mutation is GNAQ11 and BAP1 genes). CONCLUSION: Cytological diagnosis of melanoma by fine-needle aspiration biopsies and imprints from the tumor mass is a highly informative method that allows diagnosing melanoma and verifying the tumor subtype. The obtained results indicate tumor heterogeneity and differences in mutational status depending on the location of melanomas. The molecular classification of melanoma is important when choosing individualized therapy.

Methodological and TNM Focus-Based Comparison of EGFR Mutation Status in Non-Small-Cell Lung Carcinomas.

Epidermal growth factor receptor (EGFR) mutations in non-small-cell lung carcinomas (NSCLC) are a frequent class of driver mutations, and tyrosine kinase inhibitor (TKI) therapy provides considerable clinical benefits. Using the most effective and also easiest method for EGFR analysis is cost-effective and time-saving. In this study, we aimed to determine which method could be more effective by comparing the incidences of EGFR mutations in cytological and histological samples which were obtained by different methods also, whether there was a difference in the incidences of EGFR mutations between the primary foci, mediastinal lymph nodes, and distant metastatic foci. We retrospectively reviewed 420 cases of cytological materials, small biopsies, and surgical samples reported as NSCLC underwent EGFR analysis in our department between 2016 and 2022. We collected the data and interpreted the results from two different perspectives. We identified 36 EGFR mutations in 362 biopsies (9.94%) and 17 in 58 cytology samples (29.31%). There is a significant difference between the two methods (P = 0.01*). We observed 38 EGFR mutations in 320 primary foci (11.87%), 7 EGFR mutations in 36 mediastinal or subcarinal lymph nodes (19.44%), and 8 EGFR mutations in 64 distant metastatic foci (12.50%). A significant difference was also observed in pleural samples (P = 0.005*). We observed more successful results with cell blocks obtained from liquid-based cytological specimens than with formalin-fixed, paraffin-embedded tissues obtained from resection or otherwise in our clinical routine. Our study results highlight the benefits of cytological specimens in molecular treatments and current therapy modalities.

Role of Pap Smear Cervical Cytology in the Diagnosis of Extrauterine Malignancies: Largest Study of 104 Cases from Tertiary Care Cancer Centre in India

Introduction: Extrauterine malignancies in cervical samples are rarely seen. It is important to differentiate these cells from those of primary uterine malignancies to determine appropriate line of further investigations and management. Literature on these lesions is limited largely restricted to case reports. The aim of the present study was to study the spectrum and cytomorphological features of extrauterine malignancies in cervical Pap smears. Materials and Methods: It is a retrospective and descriptive study conducted in Department of Cytopathology from January 2019 to July 2023. All cases of extrauterine malignancies with available cytology material were included in this study. All cases of primary uterine malignancies, i.e., uterine corpus or cervix confirmed by clinical, radiological, and histopathological examination were excluded. Results: 104 out of 11,674 cytology Pap smears were those of extrauterine malignancy. Diagnosis of extrauterine malignancy was given in 47.1% cases, 30.9% were reported as positive for malignancy without giving the possibility of an extrauterine origin, and 22.0% were reported as atypical glandular cells only. In 56 cases where Pap smear was the first investigation which led to the diagnosis. Most common extrauterine malignancy was adenocarcinoma principally from ovarian, colorectal, and vaginal origin. Other epithelial malignancies noted were urothelial carcinoma and invasive breast carcinoma. Among non-epithelial malignancies, we reported vaginal mucosal melanoma, cutaneous melanoma, acute leukaemia, and anaplastic large cell lymphoma. Conclusion: High index of suspicion, presence of squamous and glandular elements with no atypical features, and occasional clusters of cells with marked atypia and usually no necrosis in the background are helpful cytomorphological clues to raise suspicion for extrauterine malignancy. Correlation with serology, radiology, and immunocytochemistry can help in reaching final diagnosis.

Tiempos y condiciones de almacenamiento de las muestras en anatomía patológica. Recomendaciones de la Sociedad Española de Anatomía Patológica parte 1: muestras destinadas al diagnóstico

IntroductionThe correct storage of specimens in the Pathology service is of vital importance for patient safety. However, there are no clear recommendations as regarding how long samples should be stored for a minimum period. Material and methodsA working group of the Spanish Society of Anatomic Pathology has reviewed a series of recommendations established in the literature and after two rounds of consultations and a discussion and voting phase has established a series of storage time proposals. ResultsEach of the proposals is presented with the data found in the literature and sometimes offers definitions and exceptions to the proposal. ConclusionThese recommendations, which are minimums, establish a period of at least 10 years for paraffin embedded blocks (including cell blocks), histological preparations, general cytology, pathologic cervico-vaginal cytology and electron microscopy blocks; at least 3 years for cervico-vaginal cytology, 5 years for extracted nucleic acids, at least 4 weeks for tissue in formalin and from the time of diagnosis for liquid cytology material and fluids.

Multicenter clinical trial of DNA markers for non-invasive differential diagnosis of benign and malignant melanocytic lesions.

e21550 Background: The differentiation between benign melanocytic neoplasms (nevi) and malignant melanocytic neoplasms (melanoma) presents a significant diagnostic challenge, necessitating the development of accurate and reliable diagnostic markers. This study aimed to evaluate the analytical characteristics of selected DNA markers for their efficacy in differentiating these conditions. Methods: A prospective analysis was conducted on sample groups obtained from participants of the multicenter clinical study, MoleMed. The study included 112 histological samples from 100 patients and a corresponding number of cytological samples, collected with the aim of evaluating DNA markers' sensitivity and specificity. DNA extraction followed a previously developed protocol, with concentration and quality assessed through microchip electrophoresis and real-time PCR. Molecular testing targeted mutations in "hotspots" of the BRAF and NRAS genes, as well as promoter mutations in the TERT gene, employing mutation-specific PCR amplification and Sanger sequencing for verification. Results: DNA extraction was successfully performed for 101 cytological samples, with concentrations ranging from &lt; 0.001 ng/µl to 5,508 ng/µl; the average concentration was 106.52 ng/µl, and the median was 2.37 ng/µl. BRAF mutations were found in 28 samples, with the mutation c.1799T &gt; A (p.Val600Glu, V600E) in 21 samples, c.1798_1799GT &gt; AA (p.Val600Lys, V600K) in 6 samples, and c.1799_1800TG &gt; AT (p.Val600Asp, V600D) in 1 sample. NRAS mutations were detected in 11 samples, including c.181C &gt; A (p.Gln61Lys, Q61K) in 6 samples, c.182A &gt; G (p.Gln61Arg, Q61R) in 3 samples, and c.182A &gt; T (p.Gln61Leu, Q61L) in 2 samples. No cases of simultaneous presence of more than one mutation in BRAF and NRAS "hotspots" within the same cytological sample were observed. DNA profiling of cytological material revealed signs of malignant neoplasms (MN) in 49 samples and no signs of MN in 52 samples. TERT mutations c.-124C &gt; T were detected in 21 samples, c.-146C &gt; T in 31 samples (including "borderline" cases), and c.-139_-138CC &gt; TT in 4 samples. Six samples exhibited more than one mutation, highlighting the complexity of genomic alterations in these neoplasms. Conclusions: The PCR analysis of the five DNA markers on both histological and cytological samples of human skin melanomas and nevi demonstrates promising analytical characteristics, with sensitivity and specificity exceeding 70% for all tested scenarios. These results highlight the potential utility of these DNA markers in the differential diagnosis of melanocytic neoplasms, offering a reliable, non-invasive diagnostic tool that complements traditional histopathological examination. Clinical trial information: NCT04353050 .

Utility of SHOX2 and RASSF1A gene methylation detection on the residual cytology material from endobronchial ultrasound-guided transbronchial needle aspiration.

This study aims to assess the effectiveness of Short Stature Homeobox 2 (SHOX2) and RAS Association Domain Family 1 Isoform A (RASSF1A) gene methylation detection in residual liquid-based cytology (LBC) materials from Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration (EBUS-TBNA) and investigate the diagnostic accuracy of a comprehensive diagnostic approach. Between June 2022 and May 2023, a total of 110 cases that underwent EBUS-TBNA were enrolled in the study. SHOX2 and RASSF1A genes methylation detection using the residual cytological material, LBC, and cell block (CB) were conducted for each EBUS-TBNA case. The sensitivity and specificity of cytology, CB histopathology, SHOX2, and RASSF1A methylation in diagnosing EBUS-TBNA samples were determined based on follow-up data. Among the 72 cases confirmed as pulmonary carcinomas, the methylation test yielded positive results in 24 adenocarcinoma cases, 10 squamous cell carcinoma cases, and 14 small cell carcinoma cases. The sensitivity of the comprehensive diagnosis (combining LBC, CB, and methylation detection) in distinguishing metastatic pulmonary epithelial malignancies in mediastinal and hilar lymph nodes or masses from benign lesions was higher (97.22%, 70/72) than that of morphological diagnosis alone (LBC and CB) (88.89%, 64/72; P < 0.05). SHOX2 and RASSF1A methylation detection demonstrates a high sensitivity and negative predictive value in the identification of pulmonary epithelial malignancies and holds promise as a valuable ancillary approach to enhance morphological diagnosis of EBUS-TBNA.

Detection of Oncogene Hotspot Mutations in Female NSCLC Tumor DNA and Cell-Free DNA.

Non-small-cell lung cancer (NSCLC) is the most prevalent type of lung cancer, with extensively characterized mutational spectra. Several biomarkers (such as EGFR, BRAF, KRAS gene mutations, etc.) have emerged as predictive and prognostic markers for NSCLC. Unfortunately, the quality of the available tumor biopsy and/or cytology material is not always adequate to perform the necessary molecular testing, prompting the search for alternatives. Cell-free DNA (cfDNA) found in plasma is emerging as a highly promising avenue or a supplementary method for assessing the efficacy of cancer treatments. This is especially valuable in instances where conventional biopsy specimens, like formalin-fixed, paraffin-embedded (FFPE), or freshly frozen tumor tissues prove inadequate for conducting molecular pathology analyses subsequent to the initial diagnostic procedures. By leveraging cfDNA from plasma, clinicians gain an additional tool to gauge the effectiveness of cancer therapies, thereby enhancing their ability to optimize tailored treatment strategies. In this study, 51 Lithuanian females with NSCLC were analyzed, with adenocarcinoma being the predominant pathology diagnosis in 40 cases (78%). Target mutations were identified in 38 out of 51 patients (74.5%) in tumor tissue samples, while in plasma samples, they were identified in only 10 patients' samples (19.6%). Even though we did not have enough voluminous plasma samples in our study, gene mutations were detected in plasma from ten women, three of whom were diagnosed with early stages of lung cancer (stages I and II). For these patients, the following mutations were detected: deletion in exon 19 of the EGFR gene and single nucleotide polymorphisms in the TP53 and MET genes. All other women were diagnosed with stages III or IV of lung cancer. This indicates that the later stages of cancer contribute more cfDNA in plasma, making extraction less complicated.

Tuberculosis of the thyroid gland: A rare case report

Abstract Tuberculosis (TB) of the thyroid gland is exceedingly rare, even in regions like India, where tuberculosis is prevalent, with an estimated incidence of 0.1–0.4%. While tuberculosis can affect various organs, its involvement in the thyroid, pancreas, striated, and cardiac muscle is uncommon. Diagnosis poses a challenge due to the lack of distinctive clinical features—microscopic examination revealing epithelioid granulomas, sometimes with caseous necrosis and Langhans-type giant cells, aids in diagnosis. Confirmation typically involves detecting acid-fast bacilli through staining techniques like Ziehl Neelsen (ZN), molecular diagnostic methods such as TB Gene Expert or polymerase chain reaction for Mycobacterium tuberculosis (TB PCR), and histopathological examination of excised specimens. Recently, we encountered a case involving a 21-year-old male presenting with a 2-month history of diffuse neck swelling. Clinical examination suggested thyroid swelling, confirmed by ultrasound, showing a cystic lesion with necrotic areas, suggestive of thyroiditis, accompanied by bilateral cervical lymphadenopathy. Fine Needle Aspiration Cytology revealed necrotizing granulomatous thyroiditis, prompting further investigation for tuberculosis. TB Gene Expert and ZN staining performed on fine needle aspiration cytology (FNAC) material confirmed tuberculosis, leading to anti-tuberculous treatment, which resulted in a positive response. In presenting this case, we aim to highlight the rarity of tuberculosis affecting the thyroid gland and emphasize the importance of considering minimally invasive diagnostic methods, such as FNAC and molecular techniques, before resorting to surgical interventions. This case underscores the significance of clinicians’ awareness regarding unusual tuberculosis presentations and the availability of advanced diagnostic tools to ensure timely and appropriate management.

Implementation of an ISO15189 accredited next-generation sequencing service with the fully automated Ion Torrent Genexus: the experience of a clinical diagnostic laboratory

AimsNext-generation sequencing (NGS) is integral to the delivery of personalised medicine for targeted cancer therapy. Average turnaround times (TAT) from reference laboratories with advanced expertise in sequencing are typically 2–3...

Analysis of the Differences between Bethesda Groups according to Conventional Smear and Liquid-Based Cytology Methods in Cervicovaginal Cytology: A Single-Center Experience with 165,915 Cases

Introduction: Liquid-based cytology (LBC) has replaced conventional smear (CS) in the world. In this study, through a series with a large number of cases, we aimed to make a comparison and general evaluation in all groups, primarily epithelial abnormalities, according to LBC and CS methods. This study was carried out in a private pathology laboratory located in a metropolitan city, where cytological materials sent from many clinics were examined. Material and Methods: There were 165,915 cases whose smears were examined between 2012 and 2020, most of them conventional (131,224 CS, 34,691 LBC). Cases were evaluated on the basis of the Bethesda 2014 classification and divided into sub-diagnostic categories after they were divided into two main groups as “with epithelial abnormalities” and “without.” χ2 and Fischer’s precision statistical tests were conducted using SPSS 23.0 package. In the CS process, cervical samples were obtained using an endocervical brush and a spatula. Cells were directly spread onto the slides and promptly fixed in 95% ethanol, followed by staining with the standard Papanicolaou stain. For LBC ThinPrep, cervical specimens were gathered using a cervix brush. The brush was washed in a vial and discarded. Finally, cells were isolated through vacuum filtration and transferred to the slide using air pressure. Results: Squamous cell abnormalities (atypical squamous cells of undetermined significance [ASC-US], atypical squamous cells – cannot exclude high-grade squamous intraepithelial lesion [ASC-H], low-grade squamous intraepithelial lesion [LSIL], high-grade squamous intraepithelial lesion [HSIL], squamous cell carcinoma, atypical glandular cells of undetermined significance) were reported in 5,696 (3.43%) cases. ASC (ASC-US + ASC-H)/SIL ratio (1.36/2.04) was found to be 0.67 (recommended Bethesda ratio is <3). ASC-US (p < 0.001), ASC-H (p < 0.001), and HSIL(p < 0.001) detection rate of LBC was found to be significantly higher than CS. ASC-US (1.8/1.2), ASC-H (0.08/0.008), and HSIL (0.6/0.3) case ratios of LBC/CS were found to be significantly higher in LBC. LSIL (1.72/1.66) rate was similar. Conclusion: LBC is superior to CS in detecting epithelial lesions. In addition to being used as a screening method, it is clear that it makes a great contribution to reducing cervical carcinomas due to HPV typing. Definitive comments regarding comparison of methods on reactive changes and microorganism detection are challenging. Preanalytical factors might account for these situations.

Investigation of MTAP and BAP1 staining loss and P16/CDKN2A deletion in pleural cytology specimens and its role in the diagnosis of mesothelioma.

Mesothelioma is a malignant neoplasm with a poor survival rate. We aimed to investigate the importance of BAP1, MTAP (IHC), and p16/CDKN2A homozygous deletion (FISH) in cytologic material obtained from pleural effusion sampling, which is a less invasive procedure in the diagnosis of mesothelioma. Our study discussed pleural cytology samples of cases with histopathologically proven mesothelioma diagnoses between 2017 and 2022. As the control group, materials that had pleural effusion sampling for other reasons and reactive mesothelial hyperplasia were included in the study. Cell blocks prepared from these materials were subjected to fluorescent in situ hybridization for p16/CDKN2A homozygous deletion and immunohistochemistry for BAP1 and MTAP. The specificity of the P16/CDKN2A homozygous deletion in diagnosing mesothelioma is 100%. Its sensitivity is 68.75%. The specificity of BAP1 immunohistochemical nuclear expression loss is 95%, while the sensitivity is 60%. Loss of nuclear expression of MTAP alone has the lowest specificity and sensitivity, with a specificity of 86% and a sensitivity of 43%. The highest sensitivity is reached when BAP1 loss and p16/CDKN2A homozygous deletion are evaluated together, increasing to 81%. The specificity is 95%. It has been determined that any marker alone cannot be used for a definitive mesothelioma diagnosis in pleural effusion cytological specimens; however, sensitivity increases in some combinations. The combination of BAP1 immunohistochemistry and p16/CDKN2A homozygous deletion detected by FISH, which has a higher specificity and sensitivity, can be routinely used in the diagnosis of mesothelioma under the guidance of clinical and radiologic information.

Clinical and morphological features and unsolved issues in diagnosis of aggressive forms of papillary thyroid carcinoma

Differentiated thyroid cancer (DTC) is a disease with a favorable clinical course and high survival rate compared to other epithelial tumors. The majority of DTC (up to 85 %) is represented by various subtypes of papillary thyroid cancer (PTC). Aggressive forms of PTC characterized by early lymphogenous and hematogenous metastasis, low avidity to radioiodine therapy (RIT), low relapse-free survival rates and high mortality rate compared to other types of PTC occur among them. Preoperative diagnosis of PTC is based on the results of ultrasound (US) examination and fine-needle aspiration biopsy (FNA) with cytological examination of the aspirate. At the same time, the capabilities of cytological examination in determining the histological type of PTC are limited and it does not allow to predict its aggressiveness and plan adequate treatment. Molecular genetic tests of the tumor cytological and morphological material are effective in prognosis of aggressiveness of PTC due to the determination of specific mutations in the BRAF, TERT, RAS genes and the quantitative expression of oncogenic and tumor suppressive microRNAs. Some of these indicators are already used in the morphological classification of tumors of the endocrine system. At the same time, there are contradictory data concerning the connection of the molecular genetic portrait of PTC, the clinical manifestations of its aggressiveness (extrathyroidal invasion, early metastasis, and radioiodine resistance) and its pathomorphological structure. We tried to summarize and analyze the literature data regarding the diagnosis of aggressive variants of PTC.

Identification of invasive subpopulations using spatial transcriptome analysis in thyroid follicular tumors.

Follicular tumors include follicular thyroid adenomas and carcinomas; however, it is difficult to distinguish between the two when the cytology or biopsy material is obtained from a portion of the tumor. The presence or absence of invasion in the resected material is used to differentiate between adenomas and carcinomas, which often results in the unnecessary removal of the adenomas. If nodules that may be follicular thyroid carcinomas are identified preoperatively, active surveillance of other nodules as adenomas is possible, which reduces the risk of surgical complications and the expenses incurred during medical treatment. Therefore, we aimed to identify biomarkers in the invasive subpopulation of follicular tumor cells. We performed a spatial transcriptome analysis of a case of follicular thyroid carcinoma and examined the dynamics of CD74 expression in 36 cases. We identified a subpopulation in a region close to the invasive area, and this subpopulation expressed high levels of CD74. Immunohistochemically, CD74 was highly expressed in the invasive and peripheral areas of the tumor. Although high CD74 expression has been reported in papillary and anaplastic thyroid carcinomas, it has not been analyzed in follicular thyroid carcinomas. Furthermore, the heterogeneity of CD74 expression in thyroid tumors has not yet been reported. The CD74-positive subpopulation identified in this study may be useful in predicting invasion of follicular thyroid carcinomas.