Cell Block Research Articles

Breast metastasis as the first presentation of an anorectal melanoma diagnosed on fine needle aspiration cytology: a case report

BackgroundPrimary anorectal melanoma is a rare and aggressive disease accounting for approximately 1–3% of all colorectal and anal cancers.Case reportWe present a unique case of anorectal melanoma where the primary presentation was the metastatic breast lump which led to the detection of the primary tumor. A 70-year-old elderly postmenopausal female presented with complaint of left breast lump. On fine needle aspiration cytology, May Grunwald-Giemsa and Papanicolaou stained smears examined were highly cellular and showed sheets, clusters and singly scattered highly pleomorphic plasmacytoid cells having high nucleus to cytoplasmic ratio and scant to moderate cytoplasm with enlarged eccentric round to ovoid nuclei. Immunocytochemistry performed on cell block preparation showed tumor cells strongly positive for HMB-45, S-100 and negative for ER, PR, Her2neu, chromogranin, synaptophysin and Pan-CK. A thorough work up of the patient to look for the site of primary malignancy was done and radiological findings reveled presence of an ill-defined soft tissue mass in the anal canal. Biopsy from the anal mass confirmed the diagnosis of anal melanoma with metastasis to breast.ConclusionThe present case was reported to emphasize the importance of atypical presentation where the breast lump was the primary presentation of an anal melanoma. Therefore, we should have a strong index of suspicion for metastatic tumors in case of a breast lump especially when there is unusual morphology or unexpected immunophenotype, particularly if the neoplasm is triple-negative, in patients with a previous history of other malignancies.

Cytological spectrum of testicular malignancies with histopathological correlation: A retrospective analysis.

Testicular malignancy is the most common solid organ cancer occurring in young men. The most common testicular malignancy is germ cell tumor. Extragonadal malignancies such as lymphomas are rare. Testicular fine-needle aspiration cytology (FNAC) in cancer is a bit controversial amidst fear of tumor seeding along the needle tract. Nevertheless, its largely safe, cost-effective technique providing a quick and fairly reliable diagnosis. A retrospective analysis of testicular malignancies on FNAC over a period of 9 years with cyto-histological correlation wherever possible was carried out. FNAC slides and cell blocks with immunocytochemistry wherever done were retrieved. A total of 74 cases were obtained. The age ranged from 1 year to 65 years. Infiltration by leukemia was the most common malignancy detected in pediatric population, while germ cell tumors were common amongst young adults and middle-aged men. In elderly, metastatic carcinoma, infiltration by lymphoma were identified. On FNAC, 38 cases were of leukemic infiltration, 27 of germ cell tumors (subtyped as mixed germ cell tumors-15 cases, seminoma-11 cases, and yolk sac tumor-1 case) with two cases each of non-Hodgkin lymphoma, Leydig cell tumor, metastatic adenocarcinoma, and one case each of metastatic small cell carcinoma, rhabdomyosarcoma, and malignant neoplasm. Histological correlation was available in 15/74 cases. Only 3 cases were discordant. Seeding of tumor along the needle tract was not seen. The current study deciphers the cytological spectrum of testicular malignancies on FNAC and highlights its importance as a reliable modality for a prompt diagnosis of testicular tumors guiding patient management.

A Comprehensive Review of the Newest World Health Organization (WHO) Cytopathology Reporting Systems.

World Health Organization cytopathology reporting system systems have proposed for the pancreatobiliary tract, lung, lymph node, and soft tissue aligned with the updates in the World Health Organization classification of Tumor series. Among them, the pancreatobiliary tract and lung specimen reporting system have been published recently and are now a 7-tier and 5-tier category system, respectively, without any subcategorization. World Health Organization reporting systems also encourage the application of ancillary diagnostic tests like cell block preparation, immunocytochemistry, and rapid on-site evaluation for better categorization of specimens and further management, especially in indeterminate (atypical and suspicious for malignancy) and malignant cytology categories. In this brief review, we aim to provide a brief outline of both the systems and their clinical risk-based management strategies.

The Application of Stem Cell Therapy on Type 1 Diabetes

Type 1 diabetes (T1D) is also known as insulin-dependent diabetes caused by autoimmune reactions in the patient's body. T1D is caused by the destruction of the pancreatic β cells of the patient by their immune system in the case of autoimmunity. It can lead to dehydration, frequent urination, potential damage to many organs in the body etc. If left untreated, it can lead to death. Consequently, research groups have invested a huge effort to develop possible treatments for. Currently, the existing solutions for T1D mainly include insulin therapy and immunotherapy, but the high costs of insulin therapy and the adverse reactions associated with immunotherapy limits the efficacy of these treatments and are therefore imperfect. To this day, there is no permanent cure for T1D, many study groups have proved that stem cell therapy could provide a possible solution because of its success in curing other diseases. Stem cell therapy is a treatment in which pluripotent of multipotent cells are transplanted into a patient's body in a certain way to restore normal function. The capacity do differentiate into different cell types can help stem cells stop the process of β cell destructionz, preserve remaining beta cell blocks, and promote the regeneration of primary beta cells. This review provided an overview of the existing therapies available to treat T1D, gathered information from previous clinical trials and emphasized the importance of stem cell therapies based on the current state of art.

Immunocytochemistry as an Ancillary Technique in the Cytologic Diagnosis of Synovial Sarcoma: Lock With Cell Block

Introduction: Synovial sarcomas are rare tumors accounting for approximately 5-10% of soft tissue sarcomas. For synovial sarcomas that arise at unusual locations, cytologic diagnosis is challenging despite characteristic cytomorphologic features.
 Material and Methods: The use of ancillary diagnostic procedures such as immunocytochemistry (ICC) using cell blocks and molecular genetic techniques using aspirated material help in the confirmation of diagnosis. During the period of 18 months from Nov 16 to Feb 18 we reported three cases of SS that occurred in rare locations with the help of ICC on cell blocks in 2 cases and IHC in the third case.
 Results: Cells arranged in bundles, perivascular arrangement, tight clusters, acini & singly. Cells are elongated, oval to spindle, presence of uni or bipolar cytoplasmic processes with mildly pleomorphic , hyperchromatic nuclei & fine granular chromatin. TLE1 was positive in all 3 cases.
 Conclusion: TLE1 marker is an extremely useful marker that distinguishes SS from other cytological & histological mimics.
 The objective of the present study was to call attention and also highlight the variable cytology of synovial sarcoma arising at rare locations. Furthermore, we discuss use of adjunctive ICC on cell blocks that help to lock the diagnosis.

Gastric glomus tumor on EUS-FNA-based cytology: clinicopathologic and immunohistochemical features of 4 cases, including 1 case with associated MIR143HG-NOTCH2 fusion gene.

Gastric glomus tumor (GT) is a rare submucosal tumor for which the preoperative diagnosis can be challenging. We report the cytomorphologic and immunohistochemical features of 4 gastric GTs diagnosed by endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) cytology. Files were searched to identify gastric GTs diagnosed by EUS-FNA between 2018 and 2021. A total of 4 cases of gastric GTs (3 men and 1 women; mean age, 60 years) were included. Three GTs were located in the gastric antrum and one in the gastric body. Their size ranged from 2 to 2.5 cm. Three patients presented with epigastric discomfort and one with chest wall discomfort. Rapid on-site evaluation was performed for 3 cases; the findings for all 3 were indeterminate. The smears were moderate to highly cellular and showed loose clusters of evenly distributed small- to medium-size bland tumor cells. The tumor cells had centrally located round to oval nuclei with inconspicuous nucleoli and scant to moderate amount of eosinophilic to clear cytoplasm. Examination of the cell blocks revealed branching small vessels surrounded by small- to medium-size cells. The neoplastic cells were positive for smooth muscle actin and synaptophysin and negative for AE1/AE3 and S-100. C-KIT and CD34 were variably positive. Ki-67 was <2% positive. In 1 case, the fusion panel-solid tumor (50 genes) revealed the MIR143HG-NOTCH2 fusion gene. Smears and cell block preparation revealed angiocentric sheets of uniform, small round to oval tumor cells with pale to eosinophilic cytoplasm, intermingled with endothelial cells. The differential diagnosis of gastric GTs on rapid on-site evaluation includes neuroendocrine tumors and epithelioid or spindled cell neoplasms. Immunohistochemical and molecular studies can be helpful in the preoperative diagnosis of gastric GT.

Assessment of endobronchial ultrasound-guided bronchoscopy (EBUS) intranodal forceps biopsy added to EBUS 19-gauge transbronchial needle aspiration: A blinded pathology panel analysis.

Endobronchial ultrasound-guided (EBUS) transbronchial needle aspiration (TBNA) has significantly improved the diagnostic workup for intrathoracic lymphadenopathies. More recently, EBUS intranodal forceps biopsy (IFB) has been developed in an attempt to maximize diagnostic yield by providing additional tissue. In this study, we aimed to assess the improvement of diagnostic yield with EBUS-TBNA combined with EBUS-IFB, compared to EBUS-TBNA alone. Consecutive patients who had 19-G EBUS-TBNA and EBUS-IFB from August 30, 2018, to September 28, 2021, were included. Four senior pathologists retrospectively analyzed, independently and blindly, first, only the EBUS-TBNA samples (cell block), then, at least 1 month later, both samples from EBUS-TBNA and from EBUS-IFB together. Fifty patients were included in the study and 52 lymph nodes were analyzed. Diagnostic yield was 77% (40/52) for EBUS-TBNA alone and 94% (49/52) when combined with EBUS-IFB (p = 0.023). Malignancy was diagnosed with EBUS-TBNA combined with EBUS-IFB in 25/26 cases (96%), versus 22/26 (85%) with EBUS-TBNA alone (p = 0.35); and 4/5 (80%) versus 2/5 (40%) for lymphoma specifically. Kappa interobserver agreement was 0.92 for EBUS-IFB and 0.87 for EBUS-TBNA alone. Nonmalignant condition was diagnosed with EBUS-TBNA combined with EBUS-IFB in 24/26 cases (92%), versus 18/26 (69%) for EBUS-TBNA alone (p = 0.07). The use of EBUS-IFB combined with 19-G EBUS-TBNA improves the mediastinal lymph node diagnostic yield However the benefit appears to be mainly restricted to nonmalignant histology.

Cytological Features of a Metastatic Angiosarcoma in the Lymph Node Diagnosed via Liquid-Based Cytology.

Angiosarcoma is a soft tissue sarcoma of vascular origin, with more than half of the cases arising in the skin and affecting primarily the face and scalp of elderly males. Furthermore, cutaneous angiosarcoma exhibits a higher incidence of lymph node metastases than other types of sarcomas. Angiosarcomas are rarely aspirated and are occasionally encountered on cytological samples. It is a diagnostic challenge in evaluating fine needle aspiration (FNA) from a metastatic angiosarcoma without the knowledge of prior history. We present a case of scalp angiosarcoma with disease progression to erythroderma and cervical lymphadenopathy 20 months after. FNA of the cervical node revealed vasoformative features, including hemophagocytosis, formation of an intracytoplasmic lumen/vacuole, endothelial wrapping, and cell grasping. The diagnosis of nodal metastasis by angiosarcoma was confirmed with immunohistochemistry (IHC) using two vascular markers on cell block sections. Our case demonstrates the recognizable cytomorphologic clues for this rare metastatic malignancy.

Comparison of immunohistochemical mesothelial biomarkers in paired biopsies and effusion cytology cell blocks from pleural mesothelioma.

Traditionally, the diagnosis of pleural mesothelioma is based on histological material. Minimally invasive effusion cytology specimens are an alternative that, like biopsies, require ancillary analyses. Validation of immunohistochemical (IHC) analyses on cytology, including the surrogate markers for molecular alterations BAP1 and MTAP, is of interest. IHC for eight different markers was performed on 59 paired formalin-fixed, paraffin-embedded pleural biopsies and pleural effusion cell blocks with mesothelioma. Immunoreactivity in ≥10% of tumour cells was considered positive/preserved. The concordance between histological and cytological materials was assessed. The overall percentage of agreement between the histological epithelioid component in 58 biopsies and paired cell blocks was 93% for calretinin, 98% for CK5, 97% for podoplanin, 90% for WT1, 86% for EMA, 100% for desmin, 91% for BAP1, and 72% for MTAP. For 11 cases with biphasic or sarcomatoid histology, the concordance between cytology and the histological sarcomatoid component was low for calretinin, CK5, and WT1 (all ≤45%). For the whole cohort, loss of both BAP1 and MTAP was seen in 40% while both markers were preserved in 11% of the biopsies for epithelioid histology. The corresponding numbers were 54% and 8%, respectively, for the paired cell blocks. Generally, a high concordance for IHC staining was seen between paired biopsies and pleural effusion cell blocks from mesotheliomas, but the somewhat lower agreement for WT1, EMA, and especially MTAP calls for further investigation and local quality assurance. The lower concordance for the sarcomatoid subtype for some markers may indicate biological differences.

Diagnostic Utility of Claudin4 and Comparison with BerEp4 as a Marker for Metastatic Adenocarcinoma in Serous Effusions

Introduction: Fluid cytology for malignant cells is important for diagnosis and staging of malignancies. Morphological overlap between reactive mesothelial cells and adenocarcinoma poses challenges, for which many immunohistochemical markers like BerEp4 and MOC-31 have been used extensively. Claudin4 is a new marker with promising results; however, further studies are required to establish its role as a pan-carcinoma marker in serous effusions. This study aimed to determine the utility of Claudin4 in diagnosing metastatic adenocarcinoma in effusions and comparing its performance with BerEp4. Methods: Claudin4 immunohistochemistry (IHC) was performed on effusion cell blocks (n = 60) reported as positive or suspicious for metastatic adenocarcinoma on cytology over a 1-year period and was scored for intensity (0–3) and percentage of positive cells (0–4). The results were compared with BerEp4 IHC and correlated with follow-up. Ten benign effusions were included as negative controls. Results: Claudin4 IHC was positive in all 60 (100%) cases, irrespective of the primary site. BerEp4 IHC was positive in 58 (96.7%) fluids and negative in 2 (3.3%) cases. All 10 benign effusions were negative for Claudin4 and BerEp4. Claudin4 showed higher intensity and proportion scores as compared to BerEp4 in cases where tumor cells were predominantly singly scattered and was comparable to BerEp4 where tumor cells were arranged in groups. Sensitivity, specificity, PPV, and NPV of Claudin4 in our study was 100%. Sensitivity, specificity, PPV, and NPV of BerEP4 was 96.7%, 100%, 100%, and 83.3%, respectively. Conclusion: Claudin4 IHC staining results were comparable to BerEp4, irrespective of the primary site, and it performed better in cases where tumor cells were predominantly scattered singly.

Rectovaginal endometriosis with nodular smooth muscle metaplasia diagnosed via transrectal ultrasound-guided fine-needle aspiration cytology: An underused minimally invasive diagnostic technique?

Rectovaginal endometriosis is a severe variant of deeply infiltrating endometriosis. Laparoscopic assessment with tissue sampling remains the gold standard for diagnosis of endometriosis. However, transvaginal (TVUS) and transrectal ultrasound (TRUS) have been shown to be especially helpful in the diagnosis of deep endometriosis. We present the case of a 49-year-old female with menorrhagia, dysmenorrhea, and constipation. Upon pelvic examination, an incidental mass was palpated. A computed tomography (CT) scan revealed an anterior rectal wall mass and colonoscopy was non-diagnostic. Further work-up with MRI showed a 3.9 cm mass centered within the upper rectovaginal septum. TRUS guided fine-needle aspiration (TRUS-FNA) revealed cohesive epithelial cell groups without significant cytologic atypia and a second population of bland spindle cells. Cell block slides showed glandular epithelium with associated stroma that exhibited endometrial morphology and immunophenotype. Nodular fragments of spindle cells with smooth muscle immunophenotype and fibrosis were also present. The overall morphologic findings were consistent with rectovaginal endometriosis with nodular smooth muscle metaplasia. Medical management with nonsteroidal aromatase inhibitor with radiologic follow-up was selected. Rectovaginal endometriosis represents a type of deep endometriosis usually associated with severe pelvic symptoms. Metaplastic smooth muscle cells are a frequent component of endometriosis in the rectovaginal pouch with nodular growth and may present diagnostic challenges. TRUS-FNA is a minimally invasive procedure that can provide an accurate diagnosis of endometriosis, even in this variant of deep infiltrating disease.

USE OF CYTOLOGICAL SMEAR AND CELL BLOCK TECHNIQUE FOR THE DIAGNOSIS OF PLASMA CELL MYELOMA INFILTRATION IN THE MANDIBLE

Fine-needle aspiration cytology (FNAC) is a diagnostic tool using a needle to collect samples for cytopathological examination. We report a case of a 49-year-old male patient with previous medical history of plasma cell myeloma (PCM), who had osteolytic lesions in the mandible, diagnosed by FNAC. Imaging exams exhibited multiple osteolytic lesions with ill-defined margins in the bilateral mandibular body. FNAC of the intraosseous lesion of the mandible was performed. Cytological smear and cell block technique revealed plasma cells in different degrees of maturation, predominantly eccentric pleomorphic nuclei and abundant, eosinophilic cytoplasm without a paranuclear halo. Immunohistochemistry exhibited positivity for CD138, lambda light chain and Ki-67 (proliferation index was ∼50%). Based on clinicoradiographic, microscopic, and biochemical findings, the diagnosis was PCM infiltration in the mandible. The patient underwent chemotherapy and autologous hematopoietic stem cell transplantation, and had partial remission after 10 months of follow-up. Fine-needle aspiration cytology (FNAC) is a diagnostic tool using a needle to collect samples for cytopathological examination. We report a case of a 49-year-old male patient with previous medical history of plasma cell myeloma (PCM), who had osteolytic lesions in the mandible, diagnosed by FNAC. Imaging exams exhibited multiple osteolytic lesions with ill-defined margins in the bilateral mandibular body. FNAC of the intraosseous lesion of the mandible was performed. Cytological smear and cell block technique revealed plasma cells in different degrees of maturation, predominantly eccentric pleomorphic nuclei and abundant, eosinophilic cytoplasm without a paranuclear halo. Immunohistochemistry exhibited positivity for CD138, lambda light chain and Ki-67 (proliferation index was ∼50%). Based on clinicoradiographic, microscopic, and biochemical findings, the diagnosis was PCM infiltration in the mandible. The patient underwent chemotherapy and autologous hematopoietic stem cell transplantation, and had partial remission after 10 months of follow-up.

Diagnostic Performance and Cell Count of EBUS-TBNA Needle Gauges: A Prospective Trial.

Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a well-established diagnostic procedure for evaluating hilar and mediastinal lymphadenopathies and is the gold standard for lung cancer diagnosis and staging. Recent studies assessed the effectiveness of the 19-G flex needle in obtaining larger EBUS-TBNA samples, and prospective small series gave similar results in terms of diagnostic yield when testing different gauge needles. The lack of homogeneity between series and the small sample size of some prospective cohorts poses a limit to the validity of those results. This prospective controlled study compared the 19-G flex and 22-G needles in terms of diagnostic yield. An objective laboratory method was used to count cells and compare the two needles' cytologic yields. A prospective controlled study was conducted on 90 patients undergoing EBUS-TBNA for the diagnosis of hilar and mediastinal lymphadenopathies. The institutional ethic committee (IEO573) approved the study, and informed consent was obtained from all patients. A total of 90 patients were enrolled in this study, 84.4% of whom were diagnosed with malignancy and 15.6% with non-neoplastic disease. Sensitivity for malignancy was 93.4% (CI: 87.4-97.1%) for the 19-G needle and 92.6% (CI: 86.3-96.5%) for the 22-G needle (p = 0.80). The percentage of malignant cells in the cell block was 63.9% and 61.5% for the 22-G and 19-G needles, respectively. The cell count assessed by flow cytometry was 2071 cells/µL (IQR: 600,2265) with the 22-G needle and 2761 cells/µL (IQR: 505,3250) with the 19-G needle (p = 0.79). The malignant cell count was 0.05 × 103 cells/µL with the 22-G and 0.08 × 103 cells/µL with the 19-G needle (p = 0.70). There was no difference in the presence of tissue cores in the samples, and rapid on-site evaluation (ROSE) cellularity was comparable between the two needles. The 19-G flex EBUS-TBNA needle is comparable to the 22-G needle in terms of diagnostic yield for cyto-histological evaluation of hilar and mediastinal lymphadenopathies. There is no difference between the 19-G and 22-G needle cell counts evaluated by flow cytometry.

Utility and performance of cell blocks in urine cytology: Experience at three teaching hospitals.

The use of cell block (CB) preparation is underused in urine cytology (UC) and varies among hospitals. In addition to confirming a diagnosis, CBs can be useful in cases of metastatic disease, diagnoses requiring immunohistochemical (IHC) staining, and for ancillary studies. The role of this study is to examine the performance of CBs for UC at three affiliated teaching hospitals. A retrospective review of UC cases with a CB was conducted at a county hospital, Veterans Affairs hospital, and tertiary university-based hospital. For each specimen, patient demographics, specimen type, volume, original diagnosis, and IHC stains were recorded. Each case was reviewed for diagnosis based on ThinPrep alone, diagnosis based on ThinPrep and CB, utility of CB for diagnosis, and CB cellularity. A total of 250 UC specimens with CB from 186 patients was identified. Bladder washes were the most common (72.1%). IHC stains were performed on 17.2% of cases. On blinded review, CB preparation was deemed useful in 61.2% of cases, with the highest rate for suspicious for high-grade urothelial carcinoma (SHGUC) cases (87.0%). The diagnosis based on ThinPrep review changed with incorporation of CB in 13.2% of cases, with the highest rate for SHGUC cases (43.5%). The results demonstrate that use of CB in UC confirms the final diagnosis in more than one-half of cases and changes the diagnosis in a subset of cases. Use of CB was most helpful in the SHGUC category. Further evaluation of the types of cases in which CB are prepared is warranted.

UTILITY OF HBME1 IMMUNOSTAIN IN DIFFERENTIATING REACTIVE MESOTHELIAL CELLS LESIONS FROM MALIGNANT EPITHELIAL CELLS LESIONS IN SEROUS EFFUSIONS

Objective: Parietal and the visceral layers of the mesothelium are separated by a thin layer of lubricating fluid and are made up of a single layer of flat cells. The three body cavities pleura, peritoneum, and pericardium are lined by mesothelium. The collection of fluid inside these cavities is called an effusion and may be due to a variety of inflammatory, infective, and neoplastic causes and often causes reactive mesothelial cells hyperplasia that sometimes is very hard to differentiate from malignant epithelial cells in cytopathological examinations of these fluids. Methods: We studied the utility of the Human Battifora Mesothelial Epitope-1 (HBME1) immunostain to differentiate these conditions. All the fluids from various effusions collected at the department of pathology, at a tertiary care institute in Lucknow were included in the study. Detailed history, examination findings, blood investigations, imaging findings, and histopathology reports were also noted. Fluids that showed reactive mesothelial hyperplasia and/or malignant epithelial cells on microscopic examination were further analyzed by the application of HBME1 on the cell block. Results: A total of 50 fluids were studied finally including 30 cases from the positive malignant cells group and 20 cases from the reactive mesothelial cells group. Out of 30 cases included in the malignant cells group, 16 cases (53.33%) were immunoreactive for HBME1, and out of the 20 cases included in the reactive mesothelial cell group, 18 cases (90%) showed immunoreactivity for HBME1. Conclusion: Observing this we can conclude that HBME1 immunoreactivity was significantly associated with the presence of reactive mesothelial cells compared to the malignant cells group.

First Asia-Pacific experience of trans-bronchial core biopsy with a Franseen needle.

Endobronchial ultrasound guided transbronchial needle aspiration (EBUS-TBNA) is the standard for evaluating mediastinal and hilar lesions. EBUS-TBNA is limited by small volume of material obtained for immunohistochemistry (IHC) and ancillary studies important for oncological therapies. The Franseen AcquireTM needle is designed for EBUS-transbronchial needle core biopsy (TBNB) allowing larger core sizes with evidence in gastroenterology literature but little in pulmonology. This study reports the first Asia-Pacific experience of EBUS-TBNB and adequacy of samples for diagnosis and ancillary studies. A retrospective cohort study of EBUS-TBNB at the Royal Adelaide Hospital was conducted between December 2019 and May 2021. Diagnostic rate, adequacy for ancillary studies and complications were evaluated. Samples were flushed into formalin for histological processing with no rapid on-site cytological evaluation (ROSE). For suspected lymphoma, samples were flushed into HANKS for flow cytometry. Cases performed with the Olympus VizishotTM during the same 18-month were similarly analysed. One hundred and eighty-nine patients were sampled with the AcquireTM needle. Diagnostic rate was 174/189 (92.1%). Where reported [146/189 (77.2%)], average core aggregate sample size was 13.4 mm × 10.7 mm × 1.7 mm. For non-small cell lung cancer (NSCLC) cases, 45/49 (91.8%) had adequate tissue for programmed cell death-ligand 1 (PD-L1). 32/35 (91.4%) adenocarcinoma cases had sufficient tissue for ancillary studies. There was one false negative malignant lymph node at the first AcquireTM procedure. There were no major complications. One hundred and one patients were sampled with the VizishotTM needle. Diagnostic rate was 86/101 (85.1%) with only 25/101 (24.8%) having reported tissue cores (P<0.0001 of VizishotTM) with the remaining samples processed via cell block. AcquireTM EBUS-TBNB diagnostic rate is comparable to historical data with >90% of cases having sufficient core material for ancillary studies. There appears to be a role for the AcquireTM alongside the standard of care for the work up of lymphadenopathy and particularly for lung cancer.

Description of technique for endoscopic ultrasound-guided tissue acquisition in pancreatic cancer for comprehensive molecular profiling: Results of a randomized trial.

e16306 Background: As theoretically actionable genomic lesions are found in almost 50% of pancreatic cancers which can impact clinical management in up to 30% of patients, there is increased focus on molecular profiling. Although EUS-FNB is an established method for achieving pathological diagnosis in pancreatic cancers, the technique of tissue acquisition for comprehensive molecular profiling (CMP) has not been described. Methods: Patients proven to have pancreatic adenocarcinoma using rapid onsite evaluation (ROSE) at EUS were randomized to undergo two or three dedicated FNB passes for CMP. Tissue was procured using 22G Franseen needle adopting the fanning technique and stylet-retraction maneuver. Genomic DNA and total RNA were extracted from formalin-fixed paraffin-embedded (FFPE) cell blocks (Qiagen AllPrep). Next-generation sequencing (NGS) was performed (Archer Panels) for analyzing 69 gene DNA mutations (single nucleotide variants, insertions, and deletions) and 53 RNA somatic oncogenic gene fusions. Main outcome measure was the proportion of specimens in which adequate DNA and RNA were extracted for CMP. Results: 33 patients (mean age 77.0 years [SD 7.7], female = 20) with pancreatic cancer (location = head/uncinate 22; other 11) were randomized to two (n = 17) or three (n = 16) FNB passes. While sufficient DNA was extracted from all 33 FFPE cell blocks, adequate RNA was extracted from 15 of 16 in the three pass (93.8%) vs. 16 of 17 in the two pass cohort (94.1%) (p = 0.99). There was no significant difference in the mean concentration of DNA (two passes 10.7 ng/ul [SD 7.1] vs. three passes 7.9 ng/ul [SD 4.4]; p = 0.19) or RNA (two passes 37.1 ng/ul [SD 26.5] vs. three passes 28.9 ng/ul [SD 13.2]; p = 0.29) between the groups. While somatic oncogene RNA fusion (LDAH-ETV1) predictive of metastatic disease was identified in one specimen, DNA mutations were identified in all 33 specimens that included one BRCA1 mutation that warranted Oxaliplatin-based chemotherapy. Conclusions: Specimens of adequate quantity and of high quality for comprehensive molecular profiling in pancreatic cancer can be procured in nearly 95% of patients by performing two dedicated passes using the 22G Franseen needle, adopting the fanning maneuver and stylet-retraction technique. Clinical trial information: NCT05043532 .

STAT1 negatively regulates human glioma U251 cell proliferation Huang et al: Role of STAT1 in glioma

Background Glioma is one of the most malignant tumors, which leads to high mortality in cancer patients. At present, there is no effective therapy for glioma. Therefore, it is urgent and necessary to find new molecular targets for anti-glioma therapy. Objective The present study aimed to investigate the role of signal transducer and activator of transcription 1 (STAT1) in the development and progression of human glioma and related mechanisms. Methods According to the instructions of Lipofectamine TM2000 transfection reagent, we transiently transfected the plasmid pcDNA3.1-STAT1 into glioma U251 cells. Then STAT1 expression in glioma U251 and LN382 cells was detected by Western blot. MTT was performed to assay the proliferative activity of U251 cells after STAT1 transduction, flow cytometry was used to detect cell cycle and apoptosis indicators, cell migration indicator was determined by Wound healing, and Western blot was used for detecting the expression level and change trend of p53, p21, bcl-2, Caspase-8, Cyclin A and Cyclin E in transfected cells. Results Overexpressed STAT1 significantly inhibited U251 cell proliferation and promoted U251 cell apoptosis. Meanwhile, high expression of STAT1 can increase the expression of p53, p21, and Caspase-8 while inhibiting the expression of bcl-2, Cyclin A, and Cyclin E. Conclusion Highly expressed STAT1 inhibits the proliferative activity of human glioma U251 cells and can promote tumor cell apoptosis and block cell cycle progression while regulating the expression of various signal transduction molecules. Thus, STAT1 has a critical function in the development and progression of glioma and is a novel target for glioma therapy.

The role of thyroid transcription factor-1 in differentiating lung adenocarcinomas from non-pulmonary adenocarcinoma effusions.

Effusions, characterized by abnormal fluid accumulations in body cavities, present difficulties in identifying the primary organs of metastatic tumors through cytopathologic investigation, particularly in cancer-related complications. This retrospective cross-sectional laboratory study aimed to investigate the role of thyroid transcription factor-1 (TTF-1) in distinguishing lung adenocarcinomas from non-pulmonary adenocarcinomas in effusions. The study was conducted at Almobarak Cytopathology Laboratory, a private cytopathology laboratory. H&E was used to confirm the histological diagnosis of 58 archived cell blocks. TTF-1 immunostaining patterns were then correlated with the histological diagnosis. Statistical analysis, including numerical and graphical data summaries, was conducted using the Chi-square test in SPSS 23. TTF-1 expression was observed in 20 (34.4%) cases, while 38 (65.5%) cases showed no TTF-1 reaction. Positive TTF-1 was found in pleural fluid in 61.1 % of lung adenocarcinomas, while negative TTF-1 was found in only 3.4%. TTF-1 was not detected in the majority of peritoneal fluid samples. There was a highly significant relationship between pleural fluid, TTF-1, and lung adenocarcinoma (p=0.000). The data provided further evidence that TTF-1 is a useful marker for distinguishing pulmonary adenocarcinomas from non-pulmonary adenocarcinoma tumors.

Waldenström macroglobulinemia diagnosed by bronchoalveolar lavage.

An 88-year-old woman with no significant medical history was admitted to the emergency room with persistent dyspnea and cough. Chest computed tomography (CT) showed diffuse ground-glass opacities in both the lungs (Fig. 1A). Collagen disease and infection were ruled out, and lymphoproliferative disease was suspected based on high level of WBC 15 200/μL (lymphocyte 63.7%), soluble interleukin 2 receptor 3070 (151–474) IU/mL and LDH 421 (115–245) U/L. Fluorodeoxyglucose (FDG) did not accumulate in the lymph nodes throughout the body, and its accumulation as ground-glass shadows (Fig. 1B) led us to suspect a lung lesion; therefore, we performed a bronchoscopy. The patient experienced severe hypoxemia during bronchoalveolar lavage, and the biopsy was cancelled. Cell block of bronchoalveolar lavage fluid showed CD20-positive atypical lymphocytes and plasma cells, which were κ chain-positive, IgM-positive, IgG-, IgA- and λ chain-negative (Fig. 2). Blood protein fractionation showed M-protein, with increased IgM at 679 (46–260) mg/dL, and decreased IgG at 425 (870–1700) mg/dL and IgA at 8 (110–410) mg/dL. Based on these findings, the patient was diagnosed with Waldenström macroglobulinemia (WM). The patient was treated with steroids, and fever and dyspnea symptoms were relieved. The haematologist performed a bone marrow biopsy (Fig. 3), and confirmed the diagnosis of WM based on the findings of the bone marrow biopsy and flow cytometry of peripheral blood, which showed small B cells with a κ > > λ bias, negative CD5 and CD23, and IgM-κ and Bence Jones protein-κ detected by immunofixation, after which the patient received rituximab. On chest CT 120 days after starting rituximab, the ground-glass opacities in the both lungs disappeared (Fig. 4), and there has been no relapse for more than a year to date.