Scratch-induced microstructure of Ni single crystal: A FIB-TEM study.

Malignant mesothelioma (MM) is a rare yet aggressive neoplasm that arises from mesothelial cells lining the thoracic and abdominal cavities, the pericardium, and the tunica testis. Characterized by rapid progression, high invasiveness, and a poor prognosis, MM poses significant clinical challenges. SMARCB1, also referred to as INI-1, hSNF5, or BAF47, is located on chromosome 22q11.2 and serves as a critical subunit of the SWI/SNF chromatin remodeling complex. INI-1 functions as a tumor suppressor and plays a vital role in DNA damage repair and regulation of cell growth. Case Presentation: A 37-year-old Chinese male with no history of asbestos exposure but a 10-year smoking history (30 cigarettes per day) presented with a cough lasting one month and a fever persisting for one day. Computed tomography (CT) showed multiple enlarged supraclavicular lymph nodes measuring up to 30 × 15 mm bilaterally. An ultrasound-guided fine-needle aspiration (FNA) of the left supraclavicular lymph nodes was subsequently performed. The SurePath-prepared liquid-based cytopathology (LBC) using Papanicolaou-staining revealed tumor cells organized in clusters with indistinct borders. The cells exhibited abundant cytoplasm and pleomorphic round nuclei, which displayed prominent nucleoli and mitotic activity. Additionally, scattered lymphoid fragments were observed in the background. The FNA sample was formalin-fixed and paraffin-embedded for cell block; 4-μm thick sections were prepared. Histological examination using HE-stained cell block revealed a solid sheet-like tumor growth characterized by minimal stroma and the absence of fibrovascular cores. The neoplastic cells had well-defined borders, abundant eosinophilic (partially clear) cytoplasm, and round-to-oval nuclei displaying focal atypia, finely dispersed chromatin, and prominent nucleoli. Immunohistochemistry (IHC) demonstrated positive staining for CK(pan), calretinin, D2-40, and SMARCA4, whereas NapsinA, TTF-1, P40, NUT, and WT-1 yielded negative results. The confirmed loss of INI-1 expression ultimately led to the diagnosis of metastatic INI-1 deficient MM. Conclusion: This case highlights the cytomorphological features and immunophenotype of INI-1 deficient MM diagnosed through FNA. An accurate diagnosis requires a thorough clinicopathological correlation and a comprehensive IHC panel.

Development of novel anti-CDH1/E-cadherin monoclonal antibodies for versatile applications

Leucocytozoonosis is a vector-borne blood protozoal disease that affects birds worldwide. Although Egypt has a large-scale poultry industry, data on Leucocytozoon infections in broilers remain limited. This study reports the first detection of leucocytozoonosis in 24 broiler farms (180,000 birds) in El-Sharkia governorate, Egypt. The affected broiler flocks were raised in open poultry farms located near agricultural rice fields and water sources during the late summer season, which are conditions favorable for vector proliferation. Clinically, infected birds exhibited depression, anorexia, weight loss, and anemia, with mortality rates ranging from 0.5% to 2%. Postmortem examination revealed widespread hemorrhages across multiple organs and a large pool of blood in the abdominal cavity. Microscopic examination of blood smears and cell block preparations showed the presence of various gametocytes within leukocytes and erythrocytes. Histopathological examination identified characteristic megaloschizonts in multiple organs, including the kidneys, liver, lungs, muscles, heart, intestine, pancreas, and spleen, accompanied by hemorrhages, necrosis, and mononuclear cell infiltration, primarily lymphocytes and macrophages. Molecular identification using real-time PCR confirmed L. caulleryi infection, corroborating the hematological and pathological findings. This study provides key insights into L. caulleryi diagnosis, promotes awareness among veterinarians and poultry producers, and highlights the need for improved biosecurity.

Interpretable multi-scale deep learning to detect malignancy in cell blocks and cytological smears of pleural effusion and identify aggressive endometrial cancer.

Molecular analysis of FGFR2 aberrant transcripts became crucial for clinical stratification of intrahepatic cholangiocarcinoma (iCCA) patients. Several strategies, including fluorescent in situ hybridization (FISH) and next generation sequencing (NGS), are commonly used to investigate FGFR aberrations. Here, we evaluated the technical performance of clinically implemented diagnostic strategies in 8 referral Italian institutions on artificial reference formalin-fixed paraffin-embedded (FFPE) samples. Each participating institution was requested to apply its own diagnostic testing strategy on 8 sections obtained from artificial reference specimens built to harbor FGFR3(17)-TACC3(11) rearrangement and unbalanced FGFR2. A second-round slide set hosting FGFR2(17)-BICC1(3) aberrant transcript was shared to detect clinically relevant FGFR2 fusion. Artificial reference sample was previously validated by the University of Naples Federico II before arranging the shipment. Technical procedures (e.g. extraction methods, testing platforms and assays) were recorded. Overall, cell resuspension yielded higher amounts of DNA and RNA (SNU16 61.5 ng/µl, 38100.0 pg/µl; RT112 118.0/µl, 2140.0 pg/µl, respectively) in comparison with SNU16+ RT112 mixing cell block (0.7 ng/µl DNA and 412.0 pg/µl RNA). Moreover, FFPE samples showed a higher fragmentation index (DIN 1.2 and RIN not calculated) compared with cell line resuspension (DIN 2.2 and 9.5 for SNU16 and RT112; RIN 3.9 and 6.8 for SNU16 and RT112). All participating institutions identified FGFR2(17)-BICC1(3) and FGFR3(17)-TACC3(11) aberrant transcripts. Moreover, ID#2, ID#4, ID#7 institutions also detected FGFR2(3)-CD44(1) rearrangement on RNA, whereas institutions ID#1, ID#2, ID#3, ID#5, ID#6, ID#8 identified FGFR2 CNVs on DNA. NGS represents the most suitable approach in molecular profiling of FGFR aberrant transcripts. Rings trial based on artificial reference samples play a pivotal role in optimizing routine diagnostic procedures filling the gap in clinical stratification of iCCA patients.

Evaluating the role of cytology in diagnosing primary lung carcinoma with special reference to the 2023 WHO lung cytology reporting categories.

We sought to present an alternative liquid-based cytologic (aLBC) and an agarose-based cell block (CB) preparation method for formalin-fixed fine-needle aspirations (FNAs), using breast cancer as a model, and to perform a validation procedure for immunohistochemical (IHC) assays. Between 2024 and 2025, 18 breast cancer FNA cases were collected and processed into agarose-based CB, and by applying the dropped reaspirated monolayer dried preparation method, into aLBC. Matched formalin-fixed, paraffin-embedded surgical specimens from the same patients served as gold-standard controls. Ten immunomarkers were assessed using validated IHC protocols. Human epidermal growth factor receptor 2 (HER2) status was evaluated by both IHC and dual-probe fluorescence in situ hybridization (FISH) according to 2023 American Society of Clinical Oncology/College of American Pathologists guidelines. Staining intensity, percentage of positive cells, and marker concordance were analyzed across all specimen types. Diagnostic performance metrics were calculated, and intermethod agreement was assessed using Cohen's κ coefficient (κ > 0.75 considered acceptable). The sensitivity, specificity, and either negative or positive predictive values and the accuracy values were 100% at all the tested immunostains and at the HER2 FISH assays. Accurate diagnosis from scant FNA material is an increasing demand in cytopathology. The validated aLBC and CB preparation methods proved to be cost-effective, efficient for ancillary testing, and reliable for IHC, even in low-cellularity samples.

NUT carcinoma (NC) is a rare and highly aggressive type of poorly differentiated carcinoma that originates from undefined anatomical sites, primarily in midline structures of the upper respiratory tract. We report the case of a male patient in his fifth decade of life who is a non-smoker. He presented with symptoms of dyspnea and cough, and had previously been treated for an atypical infectious process. A PET scan revealed a right pulmonary hilar mass alongside pathological lymphadenopathy. An endobronchial ultrasound-guided fine needle aspiration (EBUS-FNA) of the subcarinal lymph node was performed, followed by a cytomorphological evaluation insitu (Rapid On-Site Evaluation-ROSE) for sample management. The smear showed high hypercellularity with loosely cohesive cells arranged mostly in a single-cell pattern, exhibiting a poorly differentiated appearance. Immunohistochemical studies on the cell block indicated nuclear positivity for NUT expression, confirming the diagnosis of pulmonary NC. Poorly differentiated high-grade neoplasms in middle-aged patients present a significant challenge when the tumor sample is limited. The cytopathologist's role is to optimize the available material for a complete and accurate diagnosis, which is essential for appropriate treatment.

Fine-needle aspiration cytology (FNAC) is a vital first-line diagnostic tool in evaluating lymphadenopathy, particularly in the head and neck region, where malignancies often present with nodal enlargement. Historically, the lack of a standardised reporting system for lymph node cytology has resulted in interobserver variability and inconsistent diagnoses. The Sydney System, introduced in 2019, provided a structured five-tier framework, which was further refined by the newly proposed World Health Organisation (WHO) Reporting System for Lymph Node, Spleen, and Thymus Cytopathology (2024). This study aims to assess the diagnostic performance of the proposed Sydney system in the context of head and neck lymphadenopathy and determine the associated risk of malignancy (ROM) for each diagnostic category. A retrospective-prospective study was conducted on 295 cases of cervical and supraclavicular lymphadenopathy evaluated through FNAC, with or without ultrasound guidance, after institutional ethical clearance. ROM values were calculated and compared with those previously published in the literature for the years 2024 and 2025. Ancillary techniques, such as ROSE and cell block preparations, as well as immunohistochemistry, were employed as necessary. The study demonstrated high sensitivity (89.9%), specificity (92.9%) and diagnostic accuracy (90.47%). The ROM varied across diagnostic categories, with the highest in malignant cases and the lowest in benign cases. No cases were reported as atypia of undetermined significance (AUS), reflecting effective clinicoradiological correlation. When plotted as an ROC curve, the present study demonstrates a strong diagnostic ability (AUC≈0.91). FNAC, when interpreted using a standardised system like the WHO or Sydney framework, remains an effective, minimally invasive and cost-efficient diagnostic tool. It enhances communication, reduces diagnostic ambiguity and facilitates streamlined treatment planning, particularly in resource-limited settings.

Introduction: Although the prevalence of thyroid tumor is about 7% in the general population, the malignancy rate is approximately 5% among thyroid tumors. This emphasizes the need for effective preoperative diagnostic methods for proper patient management. Methods: A descriptive cross-sectional study was conducted at Colombo South Teaching Hospital and the Department of Pathology, University of Sri Jayewardenepura, Sri Lanka. Patients with radiologically suspected malignant thyroid nodules (n=107) were enrolled. fine needle aspiration cytology (FNAC) was performed under ultrasound scan (USS) guidance by an experienced consultant radiologist. Direct smears and cell block preparations were prepared from FNAC specimens. Smears were reviewed by two independent pathologists. USS and FNAC findings were reported according to Thyroid Imaging Reporting and Data System (TIRADS) system and Bethesda system, respectively. Descriptive statistics were expressed as frequencies and percentages. Diagnostic performance of USS and FNAC was assessed using receiver operating characteristic (ROC) curve. Sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) were calculated. Results: The Majority (79.4%) were females. Age ranged from 15 to 76 years (47.3±12.89). Malignant risk prediction of 77% was observed in TIRADS system. Malignant risk of 17.9%, 37.5%, and 87.5% were reported respectively for TIRADS 4a, 4b, 4c categories while TIRADS 5 reports a risk of 60.0%. Sensitivity, specificity, NPV, and PPV of FNAC were 88.9%, 87.5%, 96%, and 70% respectively compared with the histology results. Conclusion: Bethesda system cytology reporting and radiologic TIRADS classification are important preoperative diagnostic tools for identifying suspicious thyroid nodules in resource-limited setting. FNAC was a better diagnostic test compared to USS for evaluating thyroid nodules.

Cytology specimens are less invasive than tissue biopsies, and in some cases of non-small cell lung cancer (NSCLC), they may be the only available material. The expression of programmed death ligand-1 (PD-L1) and indoleamine 2-3 dioxygenase 2 (IDO-2) predicts the response to new treatment modalities. The aim of the study was to investigate the validity of cell blocks prepared from cytology for evaluation of PD-L1 and IDO-2 expression in NSCLC and to compare the expression of these markers in cell blocks and the corresponding tissue sample. This cross-sectional study included 32 specimens of bronchoalveolar lavage (BAL) cytology and their cell block preparations. Among them, 20 cases also had bronchoscopic biopsy. PD-L1 and IDO-2 immune staining were done for all cases. A statistically significant association is observed between high PD-L1 expression and both tumour size (p = 0.00014) and grade (p = 0.00001). High IDO-2 expression is associated with low TILs (p = 0.03). Results of PD-L1 and IDO-2 expression in bronchoscopic biopsy and cell block preparations of the same cases showed no statistical difference (p = 0.246). There was no significant difference in PD-L1 and IDO-2 expression between cytological and bronchoscopic biopsies of lung cancer, which supports the effective use of cytological specimens and reduces the need for more invasive procedures.

PurposeTo develop a manual “centrifuged-enhanced cytosmear” technique for cytologic and immunohistochemical analysis of hypocellular vitreous biopsy specimens in vitreoretinal lymphoma (VRL).MethodsDiffuse large B-cell lymphoma cells were diluted to simulate vitreous biopsy yields (1000–50,000 cells/100 µL) and fixed in PreservCyt. Samples were centrifuged in a standard laboratory centrifuge and deposited into 1000 cells/100 µL PAP pen–defined circles on charged slides. Smears were air-dried, methanol-fixed, and stained with toluidine blue and CD20 immunohistochemistry (IHC). Cell density per high-power field (40×) was quantified and compared with noncentrifuged smears. Reproducibility was assessed across replicates and between two operators. Genomic DNA concentration was measured.ResultsCentrifuged-enhanced cytosmears demonstrated significantly higher cellular densities than standard smears across all concentrations (P < 0.05), especially at 1000 to 2000 cells/100 µL, where standard smears failed to detect cells. CD20 IHC was successful without cell dropout. Genomic DNA yields ranged from 5.25 ng (1000 cells) to 143 ng (50,000 cells). Interuser variability was not significant. Comparison with cytospin clinical cases showed that centrifuged-enhanced cytosmears are of comparable cellularity to clinical cases.ConclusionsThis centrifuged-enhanced cytosmear technique reliably concentrates sparse cells for cytology and IHC. It is reproducible and cost-effective, requiring no specialized equipment. Genomic DNA yields at very low cell counts of 1000 cells/100 µL is potentially sufficient for adjuvant MYD88 mutation analysis and can be used to support cytology diagnosis.Translational RelevanceThe centrifuged-enhanced cytosmear offers an accessible alternative to cytospin preparations, enabling VRL diagnosis and IHC analysis in settings lacking cytology infrastructure or where cell blocks cannot be performed.

Although cytology has long been regarded as a traditional diagnostic tool, its development has varied across different healthcare systems. This review outlines the historical evolution and contemporary practice of cytology in Taiwan, addressing both gynecologic and non-gynecologic cytology. Cytology was first introduced to Taiwan in the 1950s with early contributions from specialists in pathology, pulmonology, gynecology, and endocrinology who established diagnostic services and training programs. These initiatives enhanced diagnostic accuracy, informed clinical management, and, through policy-driven cervical cancer screening programs, contributed to a substantial decline in disease incidence and mortality. Progress was further consolidated by the implementation of quality assurance protocols and the formal training of cytotechnologists. The adoption of internationally recognized reporting frameworks, such as the Bethesda, Paris, and Milan systems, has aligned local practice with global standards. Technological innovations, including liquid-based cytology, cell block techniques, and rapid on-site evaluation, have become integral to routine workflows, reinforcing diagnostic reliability. More recently, advances in digital cytopathology and artificial intelligence are beginning to reshape diagnostic practice and quality control.

During nervous system development, diverse types of neurons and glia are sequentially generated by self-renewing neural stem cells (NSCs). Temporal changes in gene expression within NSCs are thought to regulate neural diversity; however, the mechanisms regulating the timing of these temporal gene transitions remain poorly understood. Drosophila type 2 NSCs, like human outer radial glia, divide to self-renew and generate intermediate neural progenitors, amplifying and diversifying the population of neurons innervating the central complex, a brain region crucial for sensorimotor coordination. Type 2 NSCs express over a dozen genes temporally, broadly classified as early and late-expressed genes. A conserved gene, seven-up, mediates early to late gene expression by activating ecdysone receptor (EcR) expression. However, the timing of EcR expression and, consequently, the transition from early to late gene expression remains unknown. This study investigates whether intrinsic mechanisms of cell cycle progression and cytokinesis are required to induce the NSC early-late transition. By generating mutant clones that arrest the NSC cell cycle or block cytokinesis, we show that both processes are necessary for the early-to-late transition. When NSCs are cell cycle or cytokinesis arrested, the early gene Imp fails to be downregulated and persists in the old NSCs, while the late factors EcR and Syncrip fail to be expressed. Furthermore, we demonstrate that the early factor Seven-up is insufficient to drive the transition, despite its normal expression in cell cycle- or cytokinesis-inhibited NSCs. These results suggest that both cell-intrinsic (cell cycle/cytokinesis) and extrinsic (hormone) cues are required for the early-late NSC gene expression transition.

Incidence of malignant pleural effusion (MPE) is about 500,000 new cases per year in the United States and Europe. The major cancers associated with direct, contiguous, or hematogenous spread to pleural involvement include those from lung, breast, and hematological malignancies and is associated with a poor prognosis. The most common anomaly in thyroid development is the thyroglossal duct cyst with a possibility of malignancy within remnants are 1 to 2%, the most common being papillary adenocarcinoma. Herein, we discuss a 64 years old male patient who presented to the emergency department complaining of shortness of breath associated with chest pain eventually diagnosed as thyroglossal duct cyst papillary carcinoma with metastatic deposits in pleural fluid. Distinguishing TGD associated carcinoma from primary pyramidal thyroid lobe cancer is important. It can be difficult to diagnose preoperatively, and preoperative US-guided FNA may be considered. Immunocytochemistry on cell block sections has a great potential in arriving at a diagnosis especially in cases with unknown primary.

Objective: To investigate the clinical, cytopathological characteristics, and differential diagnosis of metastatic clear cell renal cell carcinomas (CCRCC). Methods: Nine cases of metastatic CCRCC cytologically diagnosed in the Department of Pathology, the First Affiliated Hospital of Air Force Medical University from July 2021 to December 2024 were collected. The HE staining, May-Grunewald-Giemsa staining, liquid-based slides, cell block preparation, and immunocytochemistry of EnVision two-step staining were performed. The clinical and cytopathological features, treatments and follow-up data were analyzed in combination with literature review. Results: Among the 9 cases of metastatic CCRCC, there were 7 males and 2 females. The age range was 43-78 years, and the average age was 63.6 (57.5, 72.5) years. The metastatic sites were lymph node in 3 cases (2 cases of mediastinal lymph nodes and 1 case of left cervical lymph node), bone in 3 cases (pubis, thoracic vertebrae and femur, respectively), thyroid in 2 cases, and adrenal gland, lung and pancreas in 1 case, respectively. Two of the 9 cases had two metastatic sites (case 8 had metastases of lung and mediastinal lymph nodes; case 9 had metastases of thyroid and cervical lymph nodes). The median time from the diagnosis to metastasis was 9.4 years (range 1.1 to 13.8 years). The tumor cells were arranged in papillary, acinar, sheet, cluster or single scattered pattern. Most cases had uniform nuclei with mild atypia and inconspicuous nucleoli, while some cases had variable nuclei with prominent nucleoli. The cytoplasm of the tumor cells was abundant. Some cases showed clear cytoplasm with small vacuoles, while some of them showed eosinophilic and granular cytoplasm. Immunocytochemically, the tumor cells were positive for CKpan(AE1/AE3,6/6), PAX8 (9/9), CAⅨ (9/9), CD10 (9/9), and vimiten (8/8). Patients were treated primarily with targeted therapy and/or immunotherapy and curettage and radiation therapy for bone lesions. The follow-up time ranged from 1.0 month to 41.5 months (median, 20 months), and all patients survived at the end of follow-up. Conclusions: The cytology of metastatic CCRCC often shows uniform cell size, abundant and clear cytoplasm, low nuclear/cytoplasmic ratio, and mild nuclear atypia. Its cytological diagnosis is challenging because it occurs in various sites and needs to be differentiated from primary tumors of these sites. Emphasis should be placed on the morphological recognition of CCRCC, and immunocytochemical staining should be used to improve diagnosis. When necessary, molecular testing can be employed for diagnosis. Meanwhile, the medical history should be carefully inquired by pathologists to avoid missed diagnosis and misdiagnosis.

Abstract Introduction/Objective Primary thyroid lymphoma (PTL) is rare. Most PTLs are non-Hodgkin’s lymphomas, with diffuse large B-cell lymphoma (DLBCL) being the most common type, encompassing approximately 60% to 70% of cases. Fine needle aspiration (FNA) is the initial diagnostic tool to obtain samples of thyroid nodules. It is a minimally invasive procedure and essential for the rapid diagnosis and treatment. In this report, we discuss a primary thyroid lymphoma case diagnosed by FNA. Methods/Case Report A 73-year-old man with past medical history of hypertension and hyperlipidemia presented for neck swelling. A large visible left thyroid firm mass is palpable and measures about 5cm. Ultrasonography revealed markedly asymmetrically enlarged left thyroid lobe which wrapped around trachea and extended substernally resulting in right tracheal deviation. There is no vascularity or microcalcifications. FNA smears revealed diffuse population of highly atypical cells with enlarged nuclei, prominent nucleoli and apoptosis in the background. Colloid is absent. By cytomorphology, it is a malignant neoplasm (Bethesda category VI). Immunostains were performed on the cell block. The tumor cells are positive for CD20 and negative for AE1/AE3, PAX-8, and CD10. Together with cytomorphology. The findings are consistent with lymphoma, favor diffuse large B cell lymphoma (DLBCL). Results NA Conclusion PTL is uncommon and can be confused with other thyroid malignancies like anaplastic thyroid carcinoma. It is important to consider PTL in the differential diagnoses for rapid growing thyroid nodules. FNA plays an important role as the initial diagnostic tool for primary thyroid lymphoma to avoid unnecessary surgery and initiate rapid appropriate treatment. Immunostains on the cell block can help confirm the diagnosis and further classify the subtypes.

BackgroundThe plasma-thrombin method, which uses expired blood bank plasma as an ancillary component, has been widely used in cell block (CB) preparation. However, the application of expired blood bank plasma raises concerns about nucleic acid contamination. This study investigated the feasibility of using patient-derived plasma as a substitute for blood bank plasma in the modified plasma-thrombin (MPT) method for CB preparation in endobronchial ultrasound–guided transbronchial needle aspiration (EBUS-TBNA) samples.MethodsA prospective study was conducted to compare the adequacy of CB preparation between a previously used self-clotting (SC) method and the MPT method. The EBUS-TBNA specimens from each targeted lesion were divided into paired samples: one processed using the SC method and the other using the MPT method, substituting the blood bank plasma with patient-derived plasma.ResultsA total of 82 paired EBUS-TBNA samples from 59 patients were analyzed. The diagnostic yield of the SC method and the MPT method was 86.6% and 97.6%, respectively. Among patients diagnosed with non–small cell lung cancer, the adequacy rate for molecular testing was 79.2% with the SC method and 91.7% with the MPT method.ConclusionsThe MPT method significantly improved the cellular yield of EBUS-TBNA–derived CBs. Using patient-derived fresh plasma rather than expired blood bank plasma avoids a known contamination risk. The additional step modestly prolongs the procedure and introduces minimal risks by vein puncture. This approach is generally considered cost-effective.

Tips and tricks for the cytological management of cysts