Cytological Diagnosis Research Articles

FAM19A4 and hsa-miR124-2 Double Methylation as Screening for ASC-H- and CIN1 HPV-Positive Women.

The DNA methylation levels of host cell genes increase with the severity of the cervical intraepithelial neoplasia (CIN) grade and are very high in cervical cancer. Our study aims to evaluate FAM19A4 and hsa-miR124-2 methylation in Atypical Squamous cells with high-grade squamous intraepithelial lesions (ASC-H) and in CIN1, defined as low-grade squamous intraepithelial lesions (LSILs) by the Bethesda classification, as possible early warning biomarkers for managing women with high-risk HPV infections (hrHPV). FAM19A4 and hsa-miR124-2 methylation tests were conducted on fifty-six cervical screening samples from a subset of women aged 30-64 years old. Specimens were collected into ThinPrep PreservCyt Solution. Their HrHPV genotype and cytology diagnosis were known. A Qiasure (Qiagen) was used for FAM19A4 and hsa-miR124-2 methylation testing on bisulfite-converted DNA, according to the manufacturer's specifications. The reported results were hypermethylation-positive or -negative. We found that FAM194A4 and hsa-miR124-2 methylation was detected in 75% of ASC-H cases with a persistent infection of hrHPV. A total of 60% of CIN1 lesions were found to be positive for methylation, and 83.3% were when the cytology was CIN2/3. In addition, as a novelty of this pilot study, we found that combined FAM19A4 and hsa-miR124-2 methylation positivity rates (both methylated) were associated with the HPV genotypes 16, 18, and 59 and covered 22 and 25% of ASC-H and CIN1 cases, respectively. The methylation of these two genes, in combination with HPV genotyping, can be used as an early warning biomarker in the management and follow-up of women with ASC-H and CIN1 to avoid their progression to cervical cancer.

A COMPARATIVE ANALYSIS OF THYROID CYTOLOGY CLASSIFICATION SYSTEMS: BETHESDA SYSTEM VERSUS FRCPATH, AUSTRALIAN, JAPANESE, AND ITALIAN APPROACHES IN THE INDIAN CONTEXT

Objectives: There are many different popular coding systems in use across the world such as Bethesda, FRCPath (UK), the Australian reporting system, the Japanese Thyroid Reporting System, and the Italian classification system. By evaluating the strengths and weaknesses of each system concerning the Indian patient population and health-care infrastructure, this study aims to identify the most suitable approach for accurate thyroid cytology reporting and patient management in India. Methods: A retrospective cohort study was conducted at a tertiary care center in central India following due ethical clearance. A total of 208 cases met the inclusion criteria of this study. The clinical records and the cytological records of these cases were independently analyzed and reclassified by a group of pathologists into all the above-stated thyroid coding systems. Data were collected and analyzed using Microsoft Excel 2016. Results: Approximately 88.46% of the cases were women. Medial lobe was the most common site with prevalence of 80.29%. Colloid goiter is most common cytological diagnosis. Only for 11 cases, there was a difference in coding among these coding systems. Codes 1 and 2 are same in all the coding systems and the main difference identified for cases with Bethesda Codes 3 and 5. Conclusion: The Bethesda coding is simple and reliable for coding thyroid FNAC as it has been reported over the period. The other coding systems can also be used but they will have their long learning curve. Furthermore, Bethesda is being used universally making the report understandable.

Abstract LB276: qPCR in the diagnosis of papillary thyroid cancer: Assessment of BRAF V600E mutation in FNA biopsies of thyroid nodules

Abstract Papillary thyroid carcinoma (PTC) accounts for up to 90% of thyroid cancer cases, and its differentiation from benign conditions presents a challenge in clinical practice. Fine needle aspiration biopsy (FNAB) has become a recommended practice for evaluating thyroid nodules (TN) with imaging features suspicious of malignancy. Although FNAB demonstrates high accuracy in discriminating TN, a significant number are classified as malignant indeterminate (MTI), which can only be fully diagnosed by post-surgical histologic evaluation. Up to 30% of FNAB findings are expected to be MTI, leading to a high rate of unnecessary hemithyroidectomies in about 75% of patients. In PTC, the most common mutation is c.T1799A in the BRAF gene (rs113488022), resulting in V600E in the B-raf protein, leading to an increase expression by altering the MAPK pathway. The aim of this study was to evaluate the efficacy of discriminating patients with malignant and benign TNs by detecting BRAF V600E in tissue from FNAB for molecular diagnosis of PTC. The qPCR technique with allele-specific probes was used to detect the BRAF c.1799T>A mutation in somatic DNA samples extracted by FNAB from TN (n=99) with suspected malignancy by ultrasonography. Cytologic diagnosis of biopsies was performed using the Bethesda System (TBSRTC). Diagnostic accuracy measures for evaluation of BRAF c.1799T>A as a diagnostic tool were performed with MedCalc software. Of the 99 patients, it was determined by cytology that 50 patients (50.51%) had PTC and 49 benign neoplasms (49.49%). The mutation was detected in 28 of the PTC samples (56%) (AA homozygotes [n=12, 24%] and AT heterozygotes [n=16, 32%). No mutation carriers were found among patients with benign neoplasms. Regarding accuracy measures were 56% for sensitivity of 56% and 100% specificity. The positive predictive value was 100% and the negative predictive value was 69.01%, with an accuracy of 77.78%. Finally, the negative likelihood ratio (LR-) value was 0.44 (95% CI: 0.32 to 0.60), while the positive likelihood ratio (LR+) value could not be calculated as there were no patients with the mutation in the group of patients with benign TN. In conclusion, our results suggest that the presence of BRAF c.1799T>A mutation could be indicative of PTC, confirming its usefulness in the differentiation between malignant and benign tumors. BRAF V600E detection may be valuable as an adjunct to conventional cytology, improving diagnostic accuracy and reducing the need for unnecessary surgical procedures. While highlighting the potential utility of this technique as a diagnostic tool, further analysis and consideration of other metrics is recommended for a full evaluation of its clinical relevance. Citation Format: Anette R. Gastelum-Quiroz, Andrea Ross-Orozco, Marco A. Álvarez-Arrazola, Eliakym Arambula-Meraz, Karla L. Morales-Hernández, Hanna M. Tolosa-Lerma, Fred Luque-Ortega, Paola Y. Sánchez-Aboytes, Noemí García-Magallanes. qPCR in the diagnosis of papillary thyroid cancer: Assessment of BRAF V600E mutation in FNA biopsies of thyroid nodules [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB276.

Papillae, psammoma bodies, and/or many nuclear pseudoinclusions are helpful criteria but should not be required for a definitive cytologic diagnosis of papillary thyroid carcinoma: An institutional experience of 207 cases with surgical follow up.

Noninvasive follicular thyroid neoplasm with papillary-like features (NIFTP) was introduced in 2016 replacing noninvasive follicular variant of papillary thyroid carcinoma, with recommendations to label them "noncancer." To avoid reducing risk of malignancy (ROM) and overdiagnosing NIFTP as malignant, some authors required restricted cytologic criteria (RC) for a definitive diagnosis of papillary thyroid carcinoma (PTC), including papillae, psammoma bodies. or≥3 nuclear pseudoinclusions. Since then, NIFTP criteria have been revised, biologic behavior better understood, and incidence reported to be much lower than initially anticipated. This study examines the impact of RC on PTC cytologic diagnoses, ROM, and detection of clinically significant carcinomas (CSC). A total of 207 thyroid FNAs originally diagnosed as PTC and suspicious for PTC (SPTC) with surgical follow-up were evaluated. RC were retrospectively applied to cases as a requirement for diagnosing PTC, and cases that did not meet RC were reclassified as SPTC. ROMs and diagnostic accuracies of pre- and post-RC diagnoses were correlated with followup CSC. RC were met in 118/142 (83%) and 20/65 (31%) of cases originally diagnosed as PTC and SPTC, respectively. Post-RC, 29% (19/65) of CSC originally diagnosed as SPTC were upgraded to PTC, and 17% (24/142) of CSC originally diagnosed as PTC were downgraded to SPTC. No NIFTPs were diagnosed as malignant. RC should not be required for a definitive diagnosis of PTC when other nuclear features of PTC are diffuse and overt. Applying RC, however, helps the pathologist arrive at a more definitive diagnosis of PTC in suspicious cases.

Reporting Pancreatic FNAC using the Papanicolaou System: Still a Diagnostic Challenge.

The Papanicolaou Society of Cytopathology System for reporting Pancreaticobiliary Cytology (PSCPC) is a reliable method to classify pancreatic fine needle aspiration cytology (FNAC) smears. However, it is not without practical problems which can diminish the diagnostic accuracy of the cytological diagnosis. To determine the diagnostic pitfalls while reporting cytomorphology of pancreatic lesions according to PSCPC on correlating FNAC findings with histopathology. Retrospective analysis of pancreatic FNAC smears received in the Department of Pathology of our tertiary care institute over a period of 2 years was done. The cytological diagnoses were classified according to the Papanicolaou Society of Cytopathology system of reporting pancreaticobiliary cytology and correlated with histopathology. The reasons of cyto-histological discordance were analyzed. Out of 50 cases in which both FNAC and biopsy of pancreatic lesions were done, 34 cases were positive/malignant (Category VI), eight cases were suspicious for malignancy (Category V), three cases were neoplastic (Category IV), two cases were atypical (Category III), two cases were negative for malignancy (Category II), and one case was non-diagnostic (Category I). Out of 50 cases, histopathology was non-diagnostic due to inadequate material in six cases. The cytological diagnoses were compared with histopathology in the remaining 44 cases. Categories III, IV V, and VI were considered as positive for neoplastic pathology. The sensitivity of FNAC to predict neoplastic pathology was 97.5%, while the specificity was 25%. The positive predictive value was 92.9%. Two cases reported as atypical (Category III) turned out to be adenocarcinoma on histopathology. One case reported as neuroendocrine tumor and two cases reported as adenocarcinoma on cytology displayed features of chronic pancreatitis on histology. One case reported as neoplastic mucinous cyst (Category IV) turned out to be adenocarcinoma on histology (limited concordance). The cytopathologist needs to be wary of the potential pitfalls to improve the diagnostic accuracy of FNACs.

The Difficulties and Ease of Diagnosing Anaplastic Large Cell Lymphomas on Cytology.

Anaplastic large cell lymphoma (ALCL), a unique non-Hodgkin lymphoma (NHL), is a CD30-positive neoplasm of T-cell lineage. Its distinctive yet variable cytomorphology makes diagnosing fine needle aspiration cytology (FNAC) challenging. This study was undertaken to study the cytomorphology and the utility of immunocytochemical (ICC) stains on cytology in ALCL and to discuss their morphological differential diagnosis. The present study was conducted in the Department of Pathology of a tertiary care center. A retrospective review was done from January 2017 to July 2022, and all histopathologically and immunohistochemically (IHC) diagnosed cases of ALCL were taken and correlated with the cytological diagnosis. Twenty-one cases of histopathology examination and IHC-proven cases of ALCL were retrieved from the departmental archives and reviewed. The ages ranged from 3 to 80 years (median age 28 years). Commonly noted cytomorphologic features included singly dispersed large pleomorphic cells, hallmark cells, and Reed-Sternberg-like cells. CD15, CD30, epithelial membrane antigen, and anaplastic lymphoma kinase-1 were some of the ICC stains used in this study. All 21 cases had cytology correlation. Fourteen cases had concordant cyto-histological correlation. Seven cases of histopathologically proven ALCL were reported as Hodgkin lymphoma (HL) in three, ALCL/anaplastic diffuse large B-cell lymphoma, HL/ALCL, poorly differentiated carcinoma, and NHL in one case each on cytology. ALCL has a reasonably distinct cytomorphologic appearance and ICC staining pattern, and a careful interpretation of both helps arrive at a reliable FNAC diagnosis.

Histology Findings after Two Years of Cytology/HPV Co-Testing in Germany.

Since 1 January 2020, diagnostic confirmation of abnormalities detected in the context of cytology/HPV co-testing in cervical cancer screening under the statutory health insurance scheme in women aged 35 and over has been performed according to predefined algorithms. A colposcopy is indicated even in the case of borderline/low-grade cytological changes and/or HPV persistence. In this article we compare the histology findings after primary screening examinations in 2020/21 with those from 2018/19, thus also comparing the results of two different screening approaches. Our analysis included all of the cytology, HPV, and histology results from all primary screening examinations, as well as the resulting diagnostic confirmation and curative cases, that could be obtained by 30 June 2023. In 2018/19 these comprised 650600 cytology and 1804 histology findings, and in 2020/21 there were 491450 cytology and 7156 histology findings. The absolute numbers of histology findings and the percentage ratios of these to all cytological diagnoses are presented with comparison factors. In 2020/21 there were 5.2 times more histology findings in relation to all previous cytology examinations than in 2018/19, as well as 10.6 times more biopsies, 3.8 times more conizations, and 1.2 times more hysterectomies. There was a particularly high increase in diagnostic confirmation of borderline/low-grade or only HPV-positive findings. With co-testing, 12.7 times more CIN1, 6.4 times more CIN2, and 3.5 times more CIN3 lesions were diagnosed. The proportion of biopsies without dysplasia was 7.6 times higher than in previous years. Cervical carcinomas were diagnosed 1.8 times more frequently, and endometrial carcinomas 0.7 times less frequently. More CIN lesions were found with co-testing, but the increase in histology findings of low-grade or no dysplasia was far greater than findings of CIN3. Lesions not requiring treatment accounted for 94.4% of biopsy results in 2020/21. The use of computer-assisted LBC with progression markers could reduce this.

Cytological Grading of Lymphocytic Thyroiditis and Its Correlation With Biochemical Parameters: An Experience From a Tertiary Care Center in North India.

Chronic lymphocytic thyroiditis is the most common cause of acquired hypothyroidism. The clinical management of thyroid nodules, with or without chronic lymphocytic thyroiditis, mainly depends on clinical data, ultrasonography, and fine-needle aspiration cytology (FNAC), the latter being the gold standard for the pre-surgical diagnosis of thyroidal nodules. The grading of chronic lymphocytic thyroiditis can be divided into three categories. The spectrum of the thyroid profile can be correlated to the cytological diagnosis of chronic lymphocytic thyroiditis. This study aims to study the cytomorphology of various grades of chronic lymphocytic thyroiditis and its correlation with the hormonal profile. In this study, 44 patients with a diagnosis of lymphocytic thyroiditis on FNAC were included. The cases of lymphocytic thyroiditis were graded cytomorphologically, and correlation with the thyroid hormone profile was done. The majority of the patients were between 16 and 30 years age group, with a female predominance. The majority of the patients presented with diffuse enlargement of the thyroid gland. The maximum number of cases was graded in the grade II cytological category (70.46%). A hypothyroid profile was present in 63% of patients, followed by an euthyroid profile. The majority of patients with grade II thyroiditis also had a hypothyroid profile. However, no significant association was found between cytological grading and hormonal status. Cytological grading is a clear, easy-to-use diagnostic tool for confirmation of lymphocytic thyroiditis. However, the cytological grades show no statistically significant correlation with thyroid hormonal status. Lymphocytic thyroiditis should be diagnosed with a multidisciplinary approach, as clinical features and hormonal profile when used alone may result in a missed diagnosis.

Cytomorphological features of sebaceous carcinoma of the breast.

Sebaceous carcinoma of the breast is an extremely rare histological subtype of breast cancer, with fewer than 30 cases reported to date. Because of its extremely rare histological presentation, there are few case reports that highlight its cytological findings. In this case report, the cytomorphological features of a sebaceous carcinoma of the breast are described in detail. Cytomorphological analysis revealed atypical cells presenting predominantly as loose clusters. No tubular or papillary structures were evident in the clusters and no mucin production was observed. The diagnosis of sebaceous carcinoma of the breast requires prominent sebaceous differentiation of cells. In Papanicolaou-stained smears, the differentiated tumor cells were found within the yellowish clusters. When these yellowish clusters were observed at high magnification and shifted out of focus, the sebaceous differentiation of tumor cells could be recognized. This finding is an advantage of observing Papanicolaou-stained specimens. Like previous reports, some individual cells showing sebaceous differentiation were also observed. In cases where many yellowish clusters appear, close observation of the interior of the clusters can confirm the presence of sebaceous differentiation of tumor cells and serve as a diagnostic clue for the cytological diagnosis of sebaceous carcinoma of the breast.

Label‐Free Virtual Peritoneal Lavage Cytology via Deep‐Learning‐Assisted Single‐Color Stimulated Raman Scattering Microscopy

Clinical guidelines for gastric cancer treatment recommend intraoperative peritoneal lavage cytology to detect free cancer cells. Patients with positive cytology require neoadjuvant chemotherapy instead of instant resection, and conversion to negative cytology results in improved survival. However, pathologists’ or artificial intelligence's accuracy of cytological diagnosis is disturbed by manually produced, unstandardized slides. In addition, the elaborate infrastructure makes cytology accessible to a limited number of medical institutes. This work develops CellGAN, a deep learning method that enables label‐free virtual peritoneal lavage cytology by producing virtual hematoxylin–eosin‐stained images with single‐color stimulated Raman scattering microscopy. A structural similarity loss is introduced to overcome the challenge of unsupervised virtual pathology techniques that cannot accurately present cellular structures. This method achieves a structural similarity of 0.820 ± 0.041 and a nucleus area consistency of 0.698 ± 0.102, indicating the staining fidelity outperforms the state‐of‐the‐art method. Diagnosis using virtually stained cells reaches 93.8% accuracy and substantial consistency with conventional staining. Single‐cell detection and classification on virtual slides achieve a mean average precision of 0.924 and an area under the receiver operating characteristic curve of 0.906, respectively. Collectively, this method achieves standardized and accurate virtual peritoneal lavage cytology and holds great potential for clinical translation.

Knowledge of cytology results affects the performance of colposcopy: a crossover study

ObjectiveTo determine whether knowledge of cytology affects the colposcopist’s diagnostic accuracy in the identification of cervical intraepithelial neoplasia grade 2 and worse (≥ CIN2).MethodIn this cross-over study, healthcare professionals interpreted colposcopy images from 80 patient cases with known histological diagnoses. For each case, 2 images taken with a colposcope were provided (native and after acetic acid application). Inclusion criteria consisted of women with a transformation zone type 1 or 2, who had both a cytological and histological diagnosis. Cases were distributed across two online surveys, one including and one omitting the cytology. A wash-out period of six weeks between surveys was implemented. Colposcopists were asked to give their diagnosis for each case as < CIN2 or ≥ CIN2 on both assessments. Statistical analysis was conducted to compare the two interpretations.ResultsKnowledge of cytology significantly improved the sensitivity when interpreting colposcopic images, from 51.1% [95%CI: 39.3 to 62.8] to 63.7% [95%CI: 52.1 to 73.9] and improved the specificity from 63.5% [95%CI: 52.3 to 73.5] to 76.6% [95%CI: 67.2 to 84.0]. Sensitivity was higher by 38.6% when a high-grade cytology (ASC-H, HSIL, AGC) was communicated compared to a low-grade cytology (inflammation, ASC-US, LSIL). Specificity was higher by 31% when a low-grade cytology was communicated compared to a high-grade.ConclusionsOur data suggests that knowledge of cytology increases sensitivity and specificity for diagnosis of ≥ CIN2 lesions at colposcopy. Association between cytology and histology may have contributed to the findings.

Assessment of Pre-analytical Errors and Fostering Strategies to Enhance Accurate Results and Efficient Turnaround Times in the Cytology Laboratory of a Tertiary Care Hospital.

Introduction Pre-analytical errors in cytology laboratories can significantly impact the accuracy of diagnostic results and turnaround times, ultimately affecting patient care. This article presents an evaluation of pre-analytical errors and proposes fostering strategies to enhance accuracy and efficiency in the cytology laboratory of a tertiary care hospital. The background discusses the importance of pre-analytical processes in ensuring reliable cytological diagnoses and the common errors encountered in specimen collection, handling, and transportation. Strategies for error reduction and improvement in turnaround times include staff education, standardization of procedures, utilization of appropriate collection and transport devices, implementation of quality control measures, and utilization of automation technologies. By addressing pre-analytical errors and implementing fostering strategies, cytology laboratories can optimize diagnostic accuracy, improve patient care outcomes, and enhance overall laboratory efficiency. Aims and objectives This study aims to assess the prevalence and nature of pre-analytical errors in the cytology laboratory of a tertiary care hospital to understand the extent of the issue, identify the specific factors contributing to pre-analytical errors like specimen collection, handling, and transportation processes, and pinpoint areas for improvement. It seeks to evaluate the impact of pre-analytical errors on the accuracy of cytological results and the efficiency of turnaround times, highlighting the consequences for patient care. Furthermore, the study aims to develop targeted strategies to minimize pre-analytical errors and enhance the accuracy of cytological results. Materials and methods This study was conducted at the Cytology Laboratory of our hospital from January 2023 to December 2023 after getting proper approval from the Institutional Review Board (IRB approval number 101/02/2024/PG/SRB/SMCH). It is a retrospective analytical study, and a total of 5412 samples from patients of the outpatient (OP) department, inpatient (IP) department, and community health outreach program facilities received in the cytology laboratory were analyzed during the study period. The inclusion criteria were the test samples sent specifically for cytological analysis. The samples sent for biochemical or microbiological examination were excluded. The frequency of sample distribution and rejected samples were calculated and the results were correlated. Results A total of 5,412 samples received in the cytology laboratory were analyzed during the study period. The majority of the samples were Papanicolaou smears (2,352, 43.5%), followed by fluid cytology (1,008, 18.6%) and ultrasound-guided fine-needle aspiration cytology (FNAC, 984, 18.2%). Of the total number of samples, 225 (4.16%) were repeated and 27 (0.5%) were rejected. Conclusions Pre-analytical, analytical, and post-analytical processes are the three key factors that determine the dependability and precision of cytological test results. Detecting critical alerts such as the positivity of malignancy underscores the paramount importance of result accuracy. Implementing good laboratory practices and conducting both external and internal audits can reduce the frequency of preventable errors in a cytology laboratory, thereby ensuring enhanced precision and expedited outputs.

Cytological and cytochemical diagnosis of sideroblastic anemia

Cytological and cytochemical diagnosis of sideroblastic anemia

Atypical squamous cells of undetermined significance cervical cytology in Bahrain: Reporting rates, high-risk HPV testing, and cytologic and histopathologic follow-up findings.

Atypical squamous cells of undetermined significance cytology (ASC-US) is a challenging and equivocal diagnosis for pathologists in cervical Pap smears. The data on ASC-US cervical cytology in Bahrain are deficient. This study reviewed and identified the reporting rates, high-risk human papillomavirus (HPV) testing results, histopathologic follow-up findings, and the cytologic progression of patients diagnosed with ASC-US in Bahrain. A retrospective chart review was conducted on the medical records of 23,888 women who had Pap smear test results between January 2019 and March 2022 at the main referral tertiary hospital in Bahrain to identify the ASC-US cases. High-risk HPV (hrHPV) genotype was identified, and the histopathological results of the cervical biopsy was recorded. In addition, cytologic follow-up Pap smear tests conducted within 1 year of ASC-US diagnosis were tracked to monitor the progression or regression of ASC-US. In this study, 259 out of 23,888 women reported to have ASC-UC cytology results with a reporting rate of 1.1%. The mean age of the ASC-US cases was 43 ± 11.6. Thirty percent of the ASC-US cases tested positive for hrHPV, and they were predominantly infected with the other hrHPV genotype (75%). When the histopathological changes of the cervix were analyzed, 82.1% had normal histopathologic findings. However, low-grade cervical intraepithelial neoplasia-1 was reported in 2 cases (7.1%), and high-grade cervical lesions (cervical intraepithelial neoplasia-2 and higher) were reported in 3 cases (10.7%). Regarding the cytologic follow-up, most of the ASC-US cases were negative for intraepithelial lesion or malignancy (NILM) (73.8%) or had persistent ASC-US (17.9%). The remaining minority of cases progressed into either low-grade squamous intraepithelial lesions (6%) or high-grade squamous intraepithelial lesions (2.4%). There was a statistically significant and relatively strong association between the cytology follow-up findings and the hrHPV test results (χ2 [2] = 8.869, P < 0.012, Cramer's V = 0.417). This is the first and largest study to characterize the ASC-US cytology cases in Bahrain. The reporting rates of ASC-US fell within the worldwide reported range. Although most of the ASC-US cases regressed into NILM, ASC-US showed progression into more advanced cervical cytologic diagnoses or contributed to the malignant transformation of cervical tissue, especially in the presence of hrHPV infection which was positive in 30% of the ASC-US cases. This emphasizes the role of cotesting (combination of hrHPV genotyping and Pap smear test) and the importance of colposcopic evaluation of those women with cytologic follow-up tests at 1 year of diagnosis. Implementing these recommendations in the cervical cancer screening practice in Bahrain will help in having a better management plan for women diagnosed with ASC-US cervical cytology.

Study of p40 is superior to p63 for the diagnosis of pulmonary squamous cell carcinoma on cytological smears

Background: Histology and genetically driven management are the current standard in carcinoma lung. Adequate tissue is a challenge in a few cases. While liquid biopsy is an, it’s an always preferred to get an initial cyto/histo morphological confirmation. While the immune-histo chemistry (IHC) is a proven method to differentiate the squamous vs adeno carcinoma, the role of various “immuno-cyto-chemical” makers were not studied widely. Methods: This is a retrospective (July 2018 to December 2018) for random sample collection and prospective (Jan 2019-August 2019) study conducted from in the Department of pathology, govt. chest hospital Osmania medical college, Hyderabad as a part of PG dissertation, 120 subjects with cytological diagnosis of non-small cell lung carcinoma were analyzed during this period and others were excluded. Results: Out of 120 cases 80-adenocarcinoma (ADC) and 40-squamous cell carcinoma (SCC). Most of the patients are in 61-70 years, 20 are 71-80 years, 20 between 51-60 and 10 are between 40-50 years. In this study sensitivity of p63 and p40 is equal, but specificity and positive predictive value are higher for p40 for diagnosis of SCC. Conclusions: Immuno-cyto-chemistry is still a valid option in selected cases where getting a biopsy is difficult. Our findings recommend the use of p40 immuno staining rather than p63 as a squamous cell marker.

High-throughput and simultaneous inertial separation of tumor cells and clusters from malignant effusions using spiral-contraction-expansion channels

Tumor cell clusters are regarded as critical factors in cancer pathophysiology, and increasing evidence of their higher treatment resistance and metastasis compared to single tumor cells has been obtained. However, existing cell separation methods that are designed for single tumor cells cannot be used to simultaneously purify tumor cell clusters. To address this problem, we demonstrated a microfluidic approach for the high-throughput, continuous-flow ternary separation of single tumor cells, tumor cell clusters, and WBCs from clinical pleural or abdominal effusions by coupling slanted spiral channels and periodic contraction-expansion arrays. We first systematically explored the influence of particle size and flow rate on particle focusing. The separation performance indicated that 94.0% of WBCs were removed and more than 97% of MDA-MB-231 tumor cells were recovered at a high flow rate of 3500 µL/min. Moreover, more than 90% of tumor cell clusters were effectively preserved after separation. Finally, we successfully applied our device for the ternary separation of single tumor cells, tumor cell clusters, and WBCs from different malignant effusions collected from patients with metastatic cancer. Thus, our spiral-contraction-expansion device has potential as a sample pretreatment tool for the cytological diagnosis of malignant effusions.

Improving diagnostic yield of pancreatic serous cystadenoma with cyst fluid ancillary testing, adjunct immunohistochemistry, and additional fine-needle biopsy sampling.

Fine-needle aspiration (FNA) diagnosis of pancreatic serous cystadenoma (SCA) remains challenging. This retrospective study aimed to evaluate the roles of cyst fluid ancillary testing and combined fine-needle biopsy (FNB) in improving the diagnostic yield. The authors retrospectively reviewed cytology cases that were histologically confirmed SCAs. Clinical features and FNA cyst fluid biochemical and molecular analysis results along FNB findings were reviewed. The study cohort included 31 cases from 13 male and 18 female patients with a mean age of 65. The original cytologic diagnoses were nondiagnostic (n=6, 19%), negative for malignant cells/cyst contents (n=7, 23%), atypical cells (n=3, 10%), nonmucinous cyst (n=11, 35%), and serous cystadenoma (n=4, 13%). Cyst fluid carcinoembryonic antigen (CEA) analysis was performed in 17 cases, all of which showed a low CEA level (<192 ng/mL). All 14 cases with molecular testing showed a wild-type KRAS. Inhibin immunohistochemistry was retrospectively performed on the FNA cell blocks, inhibin was positive in six of seven cases tested. In 15 cases with concurrent FNA and FNB biopsies, the diagnosis of SCA was seen in only one FNA case (7%) but 13 FNB cases (87%). This study suggests that FNA diagnosis of SCA remains challenging even with ancillary testing including cyst fluid CEA level and KRAS mutation analysis. Adjunct inhibin immunostaining may help improve the cytologic diagnosis of selective SCA cases. FNB appears superior to FNA for a definite diagnosis of SCA.

A Cytohistologic Correlation Study of Thyroid Lesions: Evaluation of Diagnostic Accuracy and Pitfalls of Fine Needle Aspiration Cytology.

Background Only about 5% of palpable thyroid nodules are malignant; the rest are entirely benign. In order to reduce the number of unnecessary treatments and properly identify situations that need surgical intervention, it is essential to distinguish between benign and malignant lesions prior to surgery. There exists a "grey zone" in thyroid cytology characterized by a significant decrease in diagnostic accuracy, making it difficult to precisely classify the lesion and leading to discrepancies. Aims and objectives The study aims to accomplish the following objectives: (1) assess the prevalence of thyroid lesions according to age and sex; (2) evaluate the accuracy of the fine needle aspiration cytology (FNAC) in diagnosing thyroid conditions; and (3) investigate the causes of cytohistological discordance within the context of this study. Materials and methods In our five-year study of thyroid lesions, 125 cases were studied for cytohistological correlation. Discrepant and likened FNAs were classified according to the diagnostic findings. A review of the cytological smears and histological sections was conducted. Results The cytological diagnoses were correlated with histopathology in 109 cases (90.83%). A total of 11 cases (09.16%) were discrepant. There were no false positives (FPs). The causes of false negative (FN) diagnoses in our study can be attributed to geographic misses and failure to recognize dual pathologies. Conclusion FNA is a very precise and time-saving technique for the diagnosis and subsequent management of palpable thyroid nodules. Patients having thyroidectomies have a much higher malignant yield, and the frequency of procedures performed on the thyroid is decreased. When FNA interpretation based on strict specimen sufficiency standards is considered along with clinical and imaging findings, the occurrence of FN and FP diagnoses is expected to decrease.

Analytical Validation of Loss of Heterozygosity and Mutation Detection in Pancreatic Fine-Needle Aspirates by Capillary Electrophoresis and Sanger Sequencing.

Pancreatic cystic disease, including duct dilation, represents precursor states towards the development of pancreatic cancer, a form of malignancy with relatively low incidence but high mortality. While most of these cysts (>85%) are benign, the remainder can progress over time, leading to malignant transformation, invasion, and metastasis. Cytologic diagnosis is challenging, limited by the paucity or complete absence of cells representative of cystic lesions and fibrosis. Molecular analysis of fluids collected from endoscopic-guided fine-needle aspiration of pancreatic cysts and dilated duct lesions can be used to evaluate the risk of progression to malignancy. The basis for the enhanced diagnostic utility of molecular approaches is the ability to interrogate cell-free nucleic acid of the cyst/duct and/or extracellular fluid. The allelic imbalances at tumor suppressor loci and the selective oncogenic drivers are used clinically to help differentiate benign stable pancreatic cysts from those progressing toward high-grade dysplasia. Methods are discussed and used to determine the efficacy for diagnostic implementation. Here, we report the analytical validation of methods to detect causally associated molecular changes integral to the pathogenesis of pancreatic cancer from pancreatic cyst fluids.

Clinical errors in the diagnostic of cervical dysplasia. Clinical cases

Introduction: The cervical screening goal is to detect and treat precancers before cancer. We wanted to understand why errors in the clinical diagnostic and management of patients with persistence of human papillomavirus (HPV); infection occur in gynecological practice. Aim: Aim of the study was to evaluate 3 clinically important complications of situations for women aged 18-45 years. The study aimed to assess the diagnostic accuracy by colposcopy visualization, cytology and biopsy results, and presence of HPV infection. We aimed to analyze the false-negative (FN) cytology diagnoses inpatients with presence of HPV, colposcopy visualization, histologically proven cervical intraepithelial neoplasia (CIN) 2-3. Material and Methods: A retrospective analysis of errors in the management of patients was carried out, in treatment methods, especially with the use of cryodestruction, colposcopy pictures, results of cytological and histological conclusion, results of examination for oncogenic types of papillomavirus were presented. Discussion: After analyzing all errors in the management of patients with persistence of papillomavirus and preliminary cryotherapy of the cervix, the main reasons that led to the progression of the disease up to cervical cancer were noted. Conclusion: In the presence of papillomavirus, cryodestruction of the cervix is not advisable, even if no cervical dysplasia was detected by histological conclusions.