Fine-needle Aspiration Material Research Articles

Utility of Conventional ZN Staining and Modified Bleach Techniques in Detection of Acid Fast Bacilli from Aspirates of Suspected Cases of Tuberculous Lymphadenitis

Introduction: Tuberculosis (TB) is a significant public health problem in many developing countries, including India. Diagnosing tuberculous lymphadenitis, in particular, poses challenges as it requires the demonstration of Acid-Fast Bacilli (AFB) using the conventional Ziehl-Neelsen (ZN) technique. The Modified Bleach Technique, which involves liquefying the aspirated specimen with sodium hypochlorite followed by centrifugation, has been described to enhance the yield of AFB, improving sensitivity and diagnostic accuracy. Aim: To highlight the role of the bleach concentration method compared to conventional ZN staining in detecting AFB in Fine-Needle Aspiration (FNA) material from clinically suspected cases of tuberculous lymphadenitis. Materials and Methods: A prospective cross-sectional study was conducted at the Department of Pathology, MVJ Medical College and Research Hospital, Bengaluru, Karnataka, India from November 2020 to December 2022. Fine needle aspiration material from 100 cases of clinically suspected tuberculous lymphadenitis was included. In each case, part of the aspirate was processed for ZN staining, while the remaining material was used for modified bleach staining and Cartridge Based Nucleic Acid Amplification Test (CBNAAT). Patient data, including age, sex, duration and site of swelling, nature of aspirate, and cytomorphological diagnosis, were documented. The data was entered into a Microsoft excel spreadsheet and analysed using the Chi-square test, with a p-value<0.05 considered statistically significant. Results: The age range of the study participants was 1-80 years. Out of 100 cases, 13 were excluded due to inadequate material. The most common lymph node site was cervical 69 (79.3%), followed by axillary 8 (9.1%) and supraclavicular 6 (6.89%). Among the 87 cases, reactive lymphadenitis was diagnosed in 36 (41.3%), suppurative lymphadenitis in 12 (13.8%), necrotising granulomatous lymphadenitis in 12 (13.8%), granulomatous lymphadenitis in 15 (17.3%), and necrotising lymphadenitis in 12 (13.8%). Out of the 100 cases included in the study, 32 cases were confirmed as tubercular lymphadenitis by CBNAAT. Among these 32 cases, conventional ZN stain was positive for AFB in only 3 cases (9%), whereas the Modified bleach stain was positive in 16 cases (50%). The results were compared with CBNAAT findings, and the modified bleach technique showed sensitivity (59.4%), specificity (100%), Positive Predictive Value (PPV) (100%), Negative Predictive Value (NPV) (80%), and Accuracy (85%). Conclusion: The Modified bleach method of ZN staining is a simple, safe, inexpensive, and easy-to-perform technique. With the bleach method, AFB was easily visible and detectable in the majority of AFB-positive cases. Additionally, the bleach method preserved the morphology of AFB better. It improves the microscopic detection of AFB, leading to a definitive diagnosis of TB on cytology.

Cytopathology of primary sclerosing mucoepidermoid carcinoma with eosinophilia of the thyroid: a multi-institutional case series and review of literature

Sclerosing Mucoepidermoid Carcinoma with Eosinophilia (SMECE) of the thyroid is an extremely rare tumor that exhibits unique histologic characteristics and is nearly always associated with lymphocytic thyroiditis (LT). However, the cytomorphologic and clinicopathologic characteristics of SMECE have only been described in rare case reports. Authors' institution laboratory information systems were searched for records of SMECE between 2012 and 2023. Literature review was performed using keywords "Sclerosing mucoepidermoid carcinoma with eosinophilia", "thyroid", and "cytopathology" to search through institution electronic library databases for relevant articles. A total of 19 cases were identified, 3 unpublished in the authors' archives and 16 in the literature which had fine needle aspiration (FNA) material or cytologic features available for review, and were comprised of 3 males and 16 females. The common cytomorphologic characteristics of SMECE included fragments or loose clusters of intermediate-type epidermoid cells in a background of prominent LT and eosinophils. Overt keratinization, mucinous cells, and extracellular mucin were not commonly encountered, resulting in diagnostic challenges, especially if eosinophils associated with epithelial cell clusters were rare. The cases were reported as "Nondiagnostic" (1 case), "Atypia of Undetermined Significance" (4 cases), "Suspicious for Malignancy" (3 case), or "Malignant" (11 cases). The clinical course of SMECE of the thyroid varied and distinct cytomorphologic characteristics in a subset of patients who experienced aggressive disease raises the possibility of different prognostic grades. Cases with keratinized squamous cells and necrosis mimic anaplastic (undifferentiated) thyroid carcinoma, but the clinical history and radiologic findings can be helpful to exclude this diagnosis.

Mimickers of neuroendocrine tumors on endoscopic ultrasound-guided fine-needle aspirate material: Need for caution.

Cytologic diagnosis of neuroendocrine tumors can be straightforward on cytologic preparations, given the classical neuroendocrine morphology and expression of neuroendocrine markers confirmed by immunohistochemistry. However, overreliance on neuroendocrine markers can lead to misdiagnosis even if individual cell features suggest a neuroendocrine tumor. We present three unusual cases, two of which were initially diagnosed as neuroendocrine tumors and the third one carried preliminary diagnosis of neuroendocrine tumor on endoscopic ultrasound-guided fine-needle aspirates. These cases subsequently turned out to be cholangioblastic cholangiocarcinoma, metastatic melanoma, and gastric glomus tumor, respectively. We suggest approaches that could have pointed us towards the correct diagnosis at the outset and discuss potential pitfalls.

Cytohistological correlation and risk stratification of salivary gland lesions using the Milan System for Reporting Salivary Gland Cytopathology: A tertiary care centre experience.

Salivary gland neoplasms account for approximately 5% of head and neck tumours. The cytomorphology of fine needle aspiration material helps determine the preoperative assessment and risk stratification. The Milan System for Reporting Salivary Gland Cytopathology (MSRSGC) was developed to improve communication between the laboratory and treatment provider. In the present study, we stratified all salivary gland lesions according to the MSRSGC and evaluated each category's concordance and risk of malignancy (ROM). This was a 5 year retrospective study. First, all cases were assigned to one of the six MSRSGC categories. Then, following cytohistological correlation, the concordance rates and ROM were calculated based on the final histopathology report. A total of 354 cases were identified, with ages ranging from 2 to 88 years and the commonest age group was the 3rd to 4th decades. Categories I, II, III, IVA, IVB, V, and VI comprised 5.37% (19/354), 26.84% (95/354), 1.13% (04/354), 51.41% (182/354), 1.98% (07/354), 1.13% (04/354), and 12.15% (43/354) of the cases, respectively. The overall concordance rate between the cytological and histopathological diagnoses was 81.25% (65 out of 80 cases), with the sensitivity, specificity, positive predictive value, and negative predictive value at 43.48%, 96.49%, 83.33%, and 80.88%, respectively. The MSRSGC provides a better objective and structured way to communicate with the health care provider. In our study, the overall concordance rate was observed in 62/80 cases, with maximum concordance seen in categories III, IVB, and V of the MSRSGC.

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

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

The Similarities and Differences between Liquid-based and Conventional Methods in Evaluation of Thyroid Fine Needle Aspiration Cytologies.

Thyroid nodules are one of the most frequent medical issues in endocrinology in our country and around the world. The appropriate evaluation of the nodule is critical in the management of patients. Fine needle aspiration cytology (FNAC) is one of the most accurate tools for evaluating these nodules. Conventional and liquid-based (LB) methods are available for thyroid FNAC. In this paper, we aim to determine the best cytological method for the evaluation of thyroid fine needle aspiration materials. Fine needle aspiration (FNA) materials obtained and prepared by conventional and LB methods from 310 patients were evaluated. These slides were compared in terms of 12 cytological parameters in a three-tiered system, and the kappa coefficient was calculated. Two slides were prepared from FNA samples, and the rest of the material was left in protective solution for LB cytology. Surepath was used as a LB technique. Conventional slides (CS) were left to dry in the air and stained with May Grunwald Giemsa. Slides prepared by two different techniques were compared in 12 cytomorphological parameters, and Kappa coefficients and correlations were calculated. Kappa statistics. In comparison of LB and CS, the highest kappa coefficient was in chromatin texture (ĸ:0.738) and inflammatory cells (ĸ: 0.482). On other parameters, fair or poor agreement was observed. Although there are some superiorities to LB cytology, co-application of these two methods is more favorable in thyroid fine needle aspiration.

Cytologic-histopathologic correlation of bethesda category III diagnosed cases in thyroid cytology and analysis according to the bethesda system for reporting thyroid cytopathology 2023

The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC) has been used by pathologists since 2010, providing a standardized language, updated in 2017 and finally in 2023. According to the latest update, category III has been divided into two subgroups: " Atypia of uncertain significance (AUS) -nuclear atypia" and "AUS-other." We aimed to analyze the risk of malignancy in patients diagnosed with category III (AUS, follicular lesion of uncertain significance (FLUS), or AUS/FLUS) and compare them based on recent changes in cytologic-histopathologic correlation. We retrospectively evaluated thyroid fine needle aspiration materials diagnosed as category III. We categorized the cases as AUS, FLUS, or AUS/FLUS. The cytopathologic and histopathologic diagnoses in the resection materials were then compared. Based on detailed microscopic descriptions in our reports, we reclassified the cases as "AUS-nuclear atypia" and "AUS-other" according to the updated 2023 TBSRTC.Our study included 73 patients, who had undergone thyroidectomy. Of these, 45.21% were reported as FLUS, 47.94% as AUS, and 6.85% as AUS/FLUS in thyroid cytology. Surgical follow-up showed no significant differences in malignancy risk, both between the FLUS and AUS groups (p=.232) and between cases diagnosed as "AUS-nuclear atypia" and "AUS-other" (p=.826). Within the indeterminate group, category III has the lowest risk of malignancy, with a predicted risk between 10-30%. However, since this group is heterogeneous, the reported malignancy risk in the literature ranges from 6-72.9%, depending on the type of atypia leading to the AUS interpretation. According to the literature, it was reported that the subgroup exhibiting the highest nuclear atypia had the highest risk of malignancy.

Prospective Evaluation of Molecular Testing of Indeterminate Thyroid Nodule Cytologies Following Diagnostic Pathway Optimization

Following optimization of ultrasound malignancy risk assessment and evaluation of local malignancy rates in each Bethesda cytologic category, we locally developed and validated ThyroSPEC, a low-cost molecular diagnostic test that detects 140 genetic alterations in residual routine fine needle aspirate material. We followed 439 consecutive patients with indeterminate thyroid nodule (ITN) from July 2020 to December 2021 with reflex ThyroSPEC testing in the context of a centralized, optimized thyroid nodule diagnostic pathway to determine the strengths and gaps in our integrated diagnostic pathway.

Thyroid Cancer Screening Using Tumor-Associated DN T Cells as Immunogenomic Markers.

BackgroundThyroid nodules are an extremely common entity, and surgery is considered the ultimate diagnostic strategy in those with unclear malignant potential. Unfortunately, strategies aiming to predict the risk of malignancy have inadequate specificity. Our group recently found that the microenvironment of thyroid cancer is characterized by an enhanced immune invasion and activated immune response mediated by double-negative T lymphocytes (DN T) (CD3+CD4-CD8-), which are believed to enable or promote tumorigenesis. In the present work, we try to use the DN T cells’ proportion in thyroid fine-needle aspiration (FNA) material as a predictor of the risk of malignancy.MethodsWe recruited 127 patients and obtained ultrasound-guided FNA samples from subjects with cytology-positive or suspicious for malignancy and from those with benign nodular goiter associated with compressive symptoms (such as dysphagia, shortness of breath, or hoarseness), Hashimoto thyroiditis, and Graves’ disease. Out of 127, we investigated 46 FNA samples of patients who underwent total thyroidectomy and for which postoperative histological diagnosis by the academic pathologists was available. We specifically measured the number of cells expressing CD3+CD4-CD8- (DN T) as a function of total CD3+ cells in FNA samples using flow cytometry. We correlated their FNA DN T-cell proportions with the pathological findings.ResultsThe DN T cells were significantly more abundant in lymphocytic infiltrates of thyroid cancer cases compared to benign nodule controls (p < 0.0001). When the DN T-cell population exceeded a threshold of 9.14%, of total CD3+ cells, the negative likelihood ratio of being cancer-free was 0.034 (96.6% sensitivity, 95% CI, 0.915–1.000, p < 0.0001). DN T cells at <9.14% were not found in any subject with benign disease (specificity 100%). The high specificity of the test is promising, since it abolishes a false-positive diagnosis and in turn unnecessary surgical procedures.ConclusionThe present study proposes DN T cells’ proportion as a preoperative diagnostic signature for thyroid cancer that with integration of RNA transcriptomics can provide a simplified technology based on the PCR assay for the ease of operation.

Comprehensive molecular profiling of pancreatic ductal adenocarcinoma in FNA, biopsy, and resection specimens.

Molecular testing to identify molecular alterations in pancreatic ductal adenocarcinoma (PDAC) has been increasingly requested because of potential therapeutic implications. In this study, we compared the performance of PDAC fine-needle aspiration (FNA), fine-needle biopsy (FNB), and resection specimens for comprehensive molecular analysis. A next-generation sequencing-based Oncomine Comprehensive Assay (OCA) was used to analyze molecular alterations in FNA, FNB, or resection specimens. We examined adequacy and success rates for completion of molecular testing and catalogued molecular alterations in these specimen types. The cohort included 23 FNA, 20 FNB, and 27 resection cases. Gene mutation or amplification analysis was successful in 18 (78%) FNA and 16 (80%) FNB specimens, whereas gene fusion assessment succeeded in 12 (52%) FNA and 12 (60%) FNB samples. All 27 (100%) resection specimens were adequate for complete OCA. There were significant differences in success rates for mutation and amplification analysis between resection and FNA or FNB specimens (P< .01) but not between FNA and FNB samples (P> .05). Manual microdissection was less likely to be performed for FNA specimens than FNB or resection specimens (P< .01). KRAS mutation was the most common mutation identified (90%), followed by mutations in TP53 (64%), CDKN2A (25%), and SMAD4 (15%) genes. Our study demonstrated similar success rates for comprehensive molecular analysis using FNA and FNB specimens of PDAC, suggesting that FNA material could serve as an alternative source for comprehensive molecular testing. The molecular alterations identified in these specimens may have potential diagnostic and therapeutic implications.

Diagnosing gastrointestinal stromal tumors: The utility of fine-needle aspiration cytology versus biopsy.

BackgroundGastrointestinal stromal tumors (GIST) are mesenchymal tumors in the intestinal tract originating from a precursor to the interstitial cells of Cajal, which plays a role in gastric motility. The preoperative diagnosis of GIST may be very important for the surgical approach or the need for neoadjuvant treatment and is often done in conjunction with molecular testing.DesignGISTs diagnosed in Stockholm between 1999 and 2019 with biopsy and/or fine‐needle aspiration (FNA) material were included. Clinical and tumor characteristics, as well as sample representability, ancillary techniques, diagnostic accuracy, and time to diagnosis, were categorized and compared.ResultsWe identified 460 diagnostic samples from 347 patients, consisting of 212 biopsies and 248 FNAs. FNA cytology had a significantly (p < 0.05) better sample representability (92% vs. 77%), diagnostic accuracy (84% vs. 76%), and shorter time to diagnosis (4.5 vs. 12.3 days on average) in comparison with biopsies. In addition, ancillary techniques such as immunochemistry and molecular analysis for KIT and PDGFRA mutations could satisfactorily be performed on FNA materials.ConclusionsThere are advantages to both biopsy and FNA cytology in diagnosing GISTs. While the significantly shorter time to diagnosis for FNA cytology can be due to institutional differences, its many strengths make it both an accurate and time‐efficient method for preoperative diagnosis of GIST.

Validation of p16 immunocytochemical staining on alcohol‐fixed aspirate smears: A single‐institutional study with review of the literature

Determining human papillomavirus (HPV) status in oropharyngeal squamous cell carcinoma can have a significant impact on treatment and clinical outcomes. Because fine-needle aspiration (FNA) is typically an initial diagnostic modality in a patient workup for primary or suspected metastatic disease, immunostaining for p16 on FNA material is a promising option to determine HPV tumor status, possibly avoiding biopsies or excisions. In this study, the authors investigated the possibility of using alcohol-fixed smears as a reliable alternative for reporting p16 status. Twenty HPV-associated tumors and 20 non-HPV-associated tumors were identified using the gold-standard histologic cutoff for positivity of ≥70% strong nuclear and cytoplasmic staining. Matched FNA specimens were identified for comparison staining, and a positive p16 result was rendered on a single aspirate smear using the same cutoff of ≥70%. On alcohol-fixed cytology smears, 16 of 20 (80%) HPV-associated tumors showed positive p16 staining in ≥70% tumor cells. Four cases showed lower level (30%-60%) nuclear and cytoplasmic staining. Nineteen of 20 (95%) non-HPV-associated tumors showed no or minimal p16 staining (0%-10%), and 1 case had a p16-equivocal cytology result. The authors performed immunocytochemical validation for p16 using alcohol-fixed smears and observed promising results, offering this technique as a potential alternative to formalin-fixed tissue in the appropriate clinical context. By using a positive staining cutoff of ≥70%, this technique offers 80% sensitivity and 95% specificity for detecting HPV-associated tumors. Although it was not performed in the current study, HPV-specific testing on available formalin-fixed, paraffin-embedded tissue should be considered in cases with equivocal or negative p16 staining.

Needle rinse cell blocks as an ancillary technique: Diagnostic and clinical utility in gastrointestinal neoplasia.

Fine-needle aspirate (FNA) cytology is often the first-choice method for diagnosing gastrointestinal nodular lesions. The FNA material can be converted to histopathology specimens by a needle rinse cell block (NRCB) technique, allowing ancillary studies to refine the cytologic diagnosis. Despite use in human pathology, NRCB has never been applied to canine or feline gastrointestinal neoplasia. This study described NRCB methodology and its diagnostic utility in specific cases of neoplastic gastrointestinal lesions. Needle rinses with saline were performed after ultrasound-guided FNAs of two intestinal lymphomas (canine and feline) and a canine gastrointestinal stromal tumor (GIST). The NRCB was prepared using the cell tube block technique and processed for paraffin embedding. Routine immunohistochemistry protocols (using CD3, PAX-5, and Ki-67 for lymphoma cases and vimentin, desmin, S-100, and KIT markers for GIST) were applied to NRCB sections, and the results were compared with matched tissue biopsies. NRCBs with adequate cell numbers, preservation, and good separation of blood were obtained. The diagnosis and immunophenotyping were confirmed in both cases of lymphoma in NRCBs. In the GIST, the immunolabeling of the neoplastic cells in NRCB was completely concordant with the tissue biopsy. The described methodology is suitable for veterinary settings, having few technical requirements and low invasiveness. The presented cases of gastrointestinal neoplasia highlight the utility of NRCBs as a platform to conduct ancillary studies and refine the cytologic diagnosis.

Ultrasound-Guided Biopsy of Suspected Salivary Gland Lymphoma in Sjögren's Syndrome.

To evaluate the safety and utility of core needle biopsy (CNB) for diagnosis of salivary gland lymphoma in Sjögren's syndrome (SS). We analyzed data from consecutive SS patients who underwent ultrasound-guided major salivary gland CNB for lymphoma diagnosis and determined whether CNB yielded an actionable diagnosis without need for further intervention. CNBs were performed in 24 patients to evaluate discrete parotid (n = 6) or submandibular (n = 2) gland masses or diffuse enlargement (n = 16; 15 parotid). One patient had 3 CNBs of the same mass. Of the 26 CNBs, 24 included flow cytometry, using CNB and/or fine needle aspirate material, and 14 targeted sonographically identified focal lesions. No patient reported complications. In the 23 patients with 1 CNB, final diagnoses were marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT; n = 6), atypical lymphoid infiltration (n = 3), benign lymphoepithelial sialadenitis (n = 9), normal gland tissue (n = 4), and lymphoepithelial cyst (n = 1). In the patient with serial CNBs, the initial one without flow cytometry was benign, but the next 2 showed atypical lymphoid infiltration. Monoclonal lymphoid infiltration was detected in 12 patients: 6 with MALT lymphoma, 3 were benign, and 3 with atypical lymphoid infiltration. Of the latter 3, 1 was treated with rituximab and 2 with expectant observation. The diagnosis changed from atypical lymphoid infiltration to MALT lymphoma in 1 patient following biopsy of inguinal adenopathy 6 months post-CNB. CNB provided actionable results and avoided open excisional biopsies in all cases. CNB is safe and useful in the evaluation of suspected salivary gland lymphoma in SS.

Incorporating cytologic adequacy assessment into precision oncology workflow using telepathology: An institutional experience

Tumor sample quality and quantity determine the success of somatic mutation analysis. Thus, a rapid on-site evaluation (ROSE) tumor cytology adequacy assessment was incorporated into the workflow of precision oncology at Weill Cornell Medicine in New York City. Optimal samples were obtained from 68 patients with metastatic cancer. Cytopathologists performed ROSE on fine-needle aspirate samples via telepathology, and subsequently core-needle biopsies were obtained. In a retrospective manner, the concordance between adequacy assessment and the success rate of the procedure was evaluated to obtain sufficient tumor tissue for next-generation sequencing (NGS). Out of the 68 procedures, 43 were documented as adequate and 25 were documented as inadequate. The diagnostic yield of adequate procedures was 100%. Adequacy evaluation predicted the success rate of molecular profiling in 40 of 43 procedures (93%; 95% CI, 80.9-98.5 procedures). The success rate of molecular testing was significantly higher in the adequate group: 93% compared with 32% in the inadequate group (P < .0005). Seven procedures that failed to provide quality material for mutational analysis and pathological diagnosis were evaluated as inadequate. Cell block provided sufficient DNA for NGS in 6 cases. In 2 cases, a core biopsy could not be performed; hence, the fine-needle aspirate material confirmed the diagnosis and was used for NGS testing. These results support the incorporation of ROSE into the workflow of precision oncology to obtain high-quality tissue samples from metastatic lesions. In addition, NGS testing of concurrent cytology specimens with adequate cellularity can be a surrogate for NGS testing of biopsy specimens.

Comparative study of conventional Ziehl−Neelsen and bleach concentration method in detecting acid-fast bacilli in fine-needle aspiration material of lymph nodes

Background: Tuberculosis (TB), specific infectious disease, affects most of the organs of the human body. Extrapulmonary TB manifests commonly as lymphadenopathy. Emergence of acquired immunodeficiency syndrome warrants the rapid diagnosis to limit its spread. Aim and Objectives: The application of bleach concentration method in detecting tubercle bacilli in the material obtained by fine-needle aspiration (FNA) of lymph nodes, evaluating sensitivity of bleach concentration method over conventional direct smear method in diagnosing tubercular lymphadenitis, detecting bacilli in the background of granuloma formation. Materials and Methods: The prospective, hospital-based, cross-sectional study was carried out in a study population of 78 patients where 15 cases got eliminated following the cytological diagnosis of malignancy and inadequacy of sample. Patients with lymphadenopathy underwent FNA after clinical examination, history taking, and informed consent. The aspirated material was stained with Leishman, H and E, conventional Ziehl−Neelsen (ZN), and bleach concentration method. Data were collected and statistical analysis done. Results: The predominant age group affected was (16–30) years with male preponderance. 44.44% cases were cytomorphologically diagnosed with tuberculous lymphadenitis. Acid-fast Bacilli (AFB) was found in 23 cases by conventional ZN method and 39 cases by bleach concentration method. Sensitivity of bleach method was 92.35% compared to 54.76% in conventional ZN method. Bacilli positivity was more in cases presenting with necrosis than with granuloma alone. Conclusion: Bleach method for detecting AFB is more sensitive than conventional ZN method. It is safe, inexpensive, easy to perform, requires no additional equipment, and safe to handle in laboratory.

Distinguishing Non-Small Cell Lung Cancer Subtypes in Fine Needle Aspiration Biopsies by Desorption Electrospray Ionization Mass Spectrometry Imaging.

Distinguishing adenocarcinoma and squamous cell carcinoma subtypes of non-small cell lung cancers is critical to patient care. Preoperative minimally-invasive biopsy techniques, such as fine needle aspiration (FNA), are increasingly used for lung cancer diagnosis and subtyping. Yet, histologic distinction of lung cancer subtypes in FNA material can be challenging. Here, we evaluated the usefulness of desorption electrospray ionization mass spectrometry imaging (DESI-MSI) to diagnose and differentiate lung cancer subtypes in tissues and FNA samples. DESI-MSI was used to analyze 22 normal, 26 adenocarcinoma, and 25 squamous cell carcinoma lung tissues. Mass spectra obtained from the tissue sections were used to generate and validate statistical classifiers for lung cancer diagnosis and subtyping. Classifiers were then tested on DESI-MSI data collected from 16 clinical FNA samples prospectively collected from 8 patients undergoing interventional radiology guided FNA. Various metabolites and lipid species were detected in the mass spectra obtained from lung tissues. The classifiers generated from tissue sections yielded 100% accuracy, 100% sensitivity, and 100% specificity for lung cancer diagnosis, and 73.5% accuracy for lung cancer subtyping for the training set of tissues, per-patient. On the validation set of tissues, 100% accuracy for lung cancer diagnosis and 94.1% accuracy for lung cancer subtyping were achieved. When tested on the FNA samples, 100% diagnostic accuracy and 87.5% accuracy on subtyping were achieved per-slide. DESI-MSI can be useful as an ancillary technique to conventional cytopathology for diagnosis and subtyping of non-small cell lung cancers.

Adenoid cystic carcinoma cytology: Salivary gland and nonsalivary gland.

Adenoid cystic carcinoma (ACCA) is an uncommon primary malignancy of salivary glands and rarely nonsalivary tissue. This study aims to evaluate the diagnostic accuracy of ACCA on fine needle aspiration (FNA) material and the associated challenges. A search on electronic pathology database from 2006 to 2016 at The Johns Hopkins Hospital found 83 cytology specimens diagnosed as ACCA, 49 with histology follow-up. Fifty-two females and 31 males were found ranging from 37 to 95 years old (mean 62.5). The tumor size was 1 to 11.5 cm (mean 3.4). FNAs were performed on 46 salivary glands (54.88%), 12 head and neck masses (14.45%), 9 lymph nodes (10.84%), 9 tracheas/lungs (10.84%), 4 vaginal/perineum/gluteal masses (4.82%), and one for each kidney, liver and abdominal/pelvic mass (1.21%). 83 FNA diagnoses revealed 3 nondiagnostics (3.61%), 20 neoplasms with unspecified features (24.10%), 30 basaloid neoplasms (36.14%), 18 ACCA (21.69%), and 12 other malignancies (14.46%). The accuracy of FNA in diagnosis of ACCA comparing to histologic follow-up in 49 cases was 87.5% sensitivity, 66.67% specificity, with 92.11% positive predictive value and 54.55% negative predictive value. The most common mimicker was pleomorphic adenoma. ACCA can be diagnosed not only in the salivary gland FNAs, but also respiratory tract, intra-abdominal, kidney, and gynecologic regions. FNA is a preferred technique to assess mass lesions. However, a diagnosis of ACCA on FNA material should be rendered with caution since there are benign and malignant neoplasms with overlapping features. Awareness of prior medical history and ancillary studies can improve the diagnosis.

Cell Block: A Tool to Improve Cytopathologic Diagnostic Value of Fine Needle Aspiration Material

Background: The various advantages of Fine-needle aspiration (FNA) biopsy/ cytology can be limited due to inadequacy of the specimen and expertise required for diagnosis. Cell blocks (CBs), though routinely used in cytology of body fluids, can also be used in FNA material with considerable increase in diagnostic accuracy. Aims &amp; objectives: To compared FNA &amp; CB as diagnostic tool and to evaluate whether cell block adds to the diagnostic accuracy of FNA.&#x0D; Methods: This is a two years prospective study carried out in the pathology department of a tertiary heath care hospital and medical college of South India which included the cases undergoing the FNAC, and followed by the biopsy. CB was prepared using Tissue coagulum clot (TCC) method. The smears and tissue sections were assessed for cellularity and adequacy for diagnosis. CBs were grouped into four diagnostic categories. FNA &amp; CB were compared with histopathology (HPE) diagnosis to calculate Sensitivity, Specificity and Diagnostic Accuracy.&#x0D; Result: A total of 195 cases were included. Cellularity and adequacy for diagnosis were higher in CB. They were diagnostically better (superior) in 13.84% (27) cases. The sensitivity (95.77%) and diagnostic accuracy (94.87%) of CB was found to be considerably higher than FNA (78.84% and 78.46% respectively).&#x0D; Conclusion: In cases of suspicious/ intermediate diagnosis or diagnosis discordant with clinical ones, by FNA; should be followed with the CB to improve the diagnostic yield &amp; to facilitate accurate diagnoses. TCC method is a simple CB method which has higher diagnostic accuracy than FNA.

The pursuit of understanding multinucleated giant cells in indeterminate and malignant thyroid cytology samples.

Multinucleated giant cell (MGC) present in a variety of non-neoplastic and neoplastic lesions of the thyroid on cytology samples. We aimed to investigate whether certain MGC features that can help us in routine cytopathology practice. One hundred and thirty seven cases thyroid fine-needle aspiration material, which was diagnosed as indeterminate or malignant according to the Bethesda categorization, were reviewed. All cytomorphological features of the MGCs were documented. These features correlated with Bethesda categories and the final diagnosis of the subsequent surgical specimen. The presence of the MGCs was identified in 42% of the samples (58 cases-total 236 MGCs). Eighty eight of them (37%) had dense cytoplasm, 148 of them had foamy cytoplasm. The mean number of dense and foamy MGC per case was higher in the malignant category (3.2 for dense MGC and 6 for foamy MGC per case). All dense MGCs were found to be in patients with papillary thyroid carcinoma (PTC). Remarkably, follicular patterned lesions had less often dense MGC compared to papillary patterned lesions (16.6% and 46.8% respectively). We detected a nuclear irregularity in 49 dense MGC (55%), groove-like features in 24 dense MGC (27.2%), and presence of some material/cell in 8 dense MGC (9%). The presence of dense MGC is an important finding for guiding accurate Bethesda categorization, especially in indeterminate and malignant categories. Besides, the presence of some material or cell in MGCs cytoplasm can be an important indicator for the prognosis of the PTC cases.