Cytological Evaluation Research Articles

Diagnostic Accuracy of Fine Needle Aspiration Cytology (FNAC) in Salivary Gland Tumors.

Professor Oral Pathology, Islamabad Dental Hospital, Islamabad 6Demonstrator Oral Pathology Margalla Institute of Health Sciences, Islamabad Author`s Contribution 1Conception and idea acquisition of data, 2Data Analysis, 3Results interpretation, 4Editing and drafting, 5Critical Revision, 6Final approval of the study to be published Funding Source: None Conflict of Interest: None Received: February 02, 2024 Accepted: May 27, 2024 Address of Correspondent Dr. Zainab Niazi Associate Professor Oral Pathology, Islamabad Dental Hospital, Islamabad dr.zainabniazi@gmail.com ABSTRACT Objectives: To evaluate the diagnostic accuracy of fine needle aspiration cytology (FNAC) by comparing its findings with the gold standard histological features of salivary gland lesions. Methodology: This retrospective study was conducted at the Department of Histopathology, Islamabad Medical and Dental Hospital (IMDC), from January 2023 to December 2023, patients presenting with salivary gland swellings. Fifty-seven patients having salivary gland swelling were included on which both histological and cytological evaluation had been done. Twenty cases were excluded because either they had lack of correlation between FNAC and histology or on view of inadequate material. All the patients were clinically evaluated clinically and FNAC was done using 10ml disposable syringe and 23/24-gauge needle without using local anesthesia. The histopathology of the surgical specimens and cytology of the FNAC specimens were compared, and the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were evaluated. Results: On FNAC, 37 cases were diagnosed cytologically, as 9 cases identified malignant and 28 as benign. Histopathological examination confirmed 9 malignant cases and 25 benign cases, whereas 3 cases were classified as false positive. The specificity and sensitivity of FNAC were 75% and 94%, respectively. Overlapping cytological features, heterogeneity, and unsampled areas were the primary factors contributing to false-positive. Conclusion: This study showed that salivary gland FNA cytology is a valuable diagnostic technique for salivary gland lesions, however due to great variation in pleomorphic adenoma, the samples must be confirmed by histopathological features.

Comparison of selected cytomorphological features of canine pemphigus foliaceus and superficial pyoderma.

Cytological detection of acantholytic keratinocytes (acantholytic cells [AC]) helps to identify canine pemphigus foliaceus (cPF) yet AC also occurs in superficial pyoderma (SP), the main differential diagnosis. To compare selected cytomorphological features of cPF and SP and to establish cytological diagnostic criteria that could differentiate cPF from SP. 40 and 51 client-owned dogs with PF and SP, respectively. Impression smears from cPF (64), impetigo (40) and exfoliative superficial pyoderma (ESP) (17) samples were stained with Romanowsky stain, randomised, blinded and evaluated by two investigators independently. The entire sample was screened (×500 or ×1000 magnification) for round (AC1), boat (AC2) and raft AC, eosinophils and bacteria. Interobserver agreements were calculated. The average number of the 10 highest ×500 fields for AC1 and AC2 was significantly higher in PF than SP (p < 0.0001; Kruskal-Wallis test). Rafts and eosinophils were more common in PF than SP (p < 0.0001; chi-square test), while bacteria were rare in PF (5%; p < 0.0001; chi-square test). Observations between the experienced and novice investigators were highly correlated. An ROC analysis identified five AC1/×500-magnification field as a suitable cut-off value for predicting PF diagnosis. This cut-off value was tested by two additional investigators, who identified sensitivity of 84%-100%, specificity of 95%-97% and accuracy of 95%-96% for the diagnosis of cPF. Criterion-based impression smear cytological evaluation can provide strong evidence to support the clinical diagnosis. Acantholytic cell morphology varies in cPF and SP, and experience can improve accuracy in cytological differentiation.

A new deep learning model with interface for fine needle aspiration cytology image-based breast cancer detection

Cytological evaluation through microscopic image analysis of fine needle aspiration cytology (FNAC) is pivotal in the initial screening of breast cancer. The sensitivity of FNAC as a screening tool relies on both image quality and the pathologist’s expertise. To enhance diagnostic accuracy and alleviate the pathologist’s workload, a computer-aided diagnosis (CAD) system was developed. A comparative study was conducted, assessing twelve candidate pre-trained models. Utilizing a locally gathered FNAC image dataset, three superior models-MobileNet-V2, DenseNet-121, and Inception-V3-were selected based on their training, validation, and testing accuracies. Further, these models underwent evaluation in four transfer learning scenarios to enhance testing accuracy. While the outcomes were promising, they left room for improvement, motivating us to create a novel deep convolutional neural network (CNN). The newly proposed model exhibited robust performance with testing accuracy at 85%. Our research concludes that the most lightweight, high-accuracy model is the one we propose. We’ve integrated it into our user-friendly Android App, “Breast Cancer Detection System,” in TensorFlow Lite format, with cloud database support, showcasing its effectiveness. Implementing an artificial intelligent (AI)-based diagnosis system with a user-friendly interface holds the potential to enhance early breast cancer detection using FNAC.

The diagnostic utility of cell block as an adjunct to routine cytology

Despite the diagnostic utility of cell block in FNAC and fluid cytology, known for better cellular yield and improved diagnostic accuracy, the technique remains to be underutilized for diagnosing neoplastic lesions.In this paper, a hospital based prospective study of 195 samples was conducted on cytological samples with the aim (a) to evaluate the diagnostic utility of cell block as an adjunct to routine cytological evaluation of aspiration and effusion fluid specimens, (b) To correlate the findings of routine cytology, cell block and routine histopathological examination wherever possible and (c) to explore the possibility of using ancillary techniques such as immunohistochemistry on cell blocks. The study revealed, most of the patients were between 51- 60 years with female preponderance. Among 195 samples, 79 (40.5%) were peritoneal fluids and 85 (43.5%) were lymph node. The diagnosis on conventional smear and cell block showed 71 (36.5%) cases and 68 (35%) cases were negative for malignancy respectively. 6 (3%) cases and 4 (2%) cases were suspicious for malignancy respectively. 118 (60.5%) cases and 123 (63%) cases were positive for malignancy respectively. Amongst the peritoneal effusion the most common primary site was ovary whereas majority of the primary site was unknown in lymph node FNAC. Peritoneal effusion showed 8 additional cases as positive for malignancy in CB preparation which were negative or suspicious for malignancy on CS. In FNAC, two additional positive case was found in CB preparation of lymph node. A Kappa value of 89.5 % for statistical correlation between Conventional smear and cell block preparation was calculated. The use of Cell block technique increases the detection of malignancy when used as an adjunct to conventional smears. Cell block technique is simple, inexpensive and reliable adjuvant to smears and it is recommended for routine cytological diagnosis and for application of immunomarkers and molecular studies.

Cytological evaluation, culture and genomics to evaluate the microbiome in healthy rabbit external ear canals.

Lop-eared rabbits may be predisposed to otitis externa (OE) as a consequence of their ear conformation. Although otoscopy, otic cytological evaluation and culture are valuable tools in dogs and cats, published data on rabbits remain lacking. This study aimed to assess the utility of otoscopy and cytological results in evaluating healthy rabbit external ear canals (EECs) and to characterise ear cytological and microbiological findings through culture techniques and metagenomic sequencing. Sixty-three otitis-free client-owned rabbits. All rabbits underwent otoscopy and ear cytological evaluation. In a subset of 12 rabbits, further bacterial and fungal culture, fungal DNA assessment and metagenomic sequencing were performed. Otic cytological results revealed yeast in 73%, cocci in 42.9% and rods in 28.6% of healthy rabbit EECs. Compared to upright-eared rabbits, lop-eared rabbits had more discharge and more bacteria per oil immersion field. Culture isolated eight different species yet metagenomic sequencing identified 36, belonging to the Bacillota (Firmicutes), Pseudomonadota and Actinomycetota phyla. Staphylococcus were the most commonly observed species with both methods. Ten of 12 rabbits were yeast-positive on cytological evaluation with only three yielding fungal growth identified as Yarrowia (Candida) lipolytica, Eurotium echinulatum and Cystofilobasidium infirmominiatum. Healthy rabbit EECs lack inflammatory cells yet can host yeast and bacteria, emphasising the need to evaluate cytological results alongside the clinical signs. Lop-ear anatomy may predispose to bacterial overgrowth and OE. Notably, yeasts may be present despite a negative culture.

Evolutionary analysis of indeterminate cytology and risk of malignancy in a thyroid nodule unit.

The objective of this study was to analyze the evolution in the diagnosis and management of indeterminate thyroid nodules over three time periods. 3020 patients with thyroid nodules underwent cytological evaluation during three periods (2006-2008, 2012-2014, 2017-2019). Distribution of diagnostic cytologies, risk of malignancy, diagnostic performance indices of FNA, and cytologic-histologic correlation in indeterminate cytologies were analyzed. only 2.2% of cytology tests were insufficient for a diagnosis. 86.9% cytologies were benign, 1.7% malignant, and 11.4% indeterminate. Indeterminate cytology rates were 15.9% (2006-2008), 10.1% (2012-2014), and 10% (2017-2019). Surgery was performed in 13% of benign cytology, result-ing in malignant histology in 2.7%. All malignant and suspicious cytologies underwent surgery: malig-nancy confirmed in 98% and 77% of cases, respectively. All 'indeterminate with atypia' cytologies (2006-2008) and Bethesda IV (2012-2014; 2017-2019) un-derwent surgery, with malignancy confirmed in 19.6%, 43.8%, and 25.7%, respectively. In the 'inde-terminate without atypia' category (2006-2008) and Bethesda III (2012-2014; 2017-2019), diagnostic surgery was performed in 57.7%, 78.6%, and 59.4%, respectively, with malignancy confirmed in 3.3%, 20.5%, and 31.6%. The FNA sensitivity was 91.6% with a negative predictive value greater than 96% in all periods. The specificity exceeded 75% in the last two periods. Bethesda system reduces indeterminate cytologies and improves the accuracy of FNA diagnosis. We reported a higher proportion of malignancy than expected in Bethesda III, underscoring the importance of having institution-specific data to guide decision-making. However, there is a need for risk stratification tools that allow for conservative management in low-risk cases.

Cytomorphological spectrum of breast fine needle aspiration cytology using the International Academy of Cytology Yokohama System and evaluation of associated risk of malignancy–A retrospective study in a tertiary care cente

BackgroundFine-needle aspiration cytology (FNAC) of the breast is a well-established technique that aids in the preoperative identification and differentiation between nonneoplastic and neoplastic breast lesions. Breast lesions can be efficiently categorized into different cytological groups using the International Academy of Cytology (IAC) Yokohama System. The aim of this study was to classify breast lesions based on the proposed Yokohama system of reporting breast cytology and identify the need for a unified system of reporting breast lesions.Materials and methodsThis retrospective descriptive study was performed in the Department of Pathology, Government Medical College, Datia, M.P. from January 2019 to June 2023. A total of 762 FNAC slides reported during this period were retrieved, re-examined and reclassified according to the IAC Yokohama System. Histopathological diagnoses were also retrieved and correlated with the cytological findings wherever available.ResultsIn the present study, a total of 762 smears of breast fine needle aspirations were included and categorized using the IAC Yokohama System. There were 3.8% cases (29/762) in the C1 category (insufficient), 71.6% cases (545/762) in the C2 category (benign), 6.3% cases (48/762) in the C3 category (atypical), 3.1% cases (24/762) in the C4 category (suspicious for malignancy) and 15.2% (116/762) cases in the C5 category (malignant).ConclusionFNAC is a reliable diagnostic technique for the evaluation and management of breast lesions prior to surgery. Implementing the IAC Yokohama System for reporting breast cytopathology enhances the accuracy of diagnosing breast lesions, reduces missed diagnoses, and offers reliable estimates of risk of malignancy. Adopting the Yokohama system is a crucial first step in boosting the overall effectiveness of breast cytology reporting among clinicians and pathologists, which will ultimately lead to improved communication and patient care.

The use of stem cells in the treatment of mastitis in dairy cows

Mastitis is a multifactorial inflammatory disease. The increase in antibiotic resistance of bacteria that cause mastitis means that cattle breeders would prefer to reduce the use of antibiotics. Recently, therapies using mesenchymal stem cells (MSCs) from various sources have gained significant interest in the development of regenerative medicine in humans and animals, due to their extraordinary range of properties and functions. The aim of this study was to analyze the effectiveness of an allogeneic stem cells derived from bone marrow (BMSC) and adipose tissue (ADSC) in treating mastitis in dairy cattle. The research material consisted of milk and blood samples collected from 39 Polish Holstein-Friesian cows, 36 of which were classified as having mastitis, based on cytological evaluation of their milk. The experimental group was divided into subgroups according to the method of MSC administration: intravenous, intramammary, and intravenous + intramammary, and according to the allogeneic stem cells administered: BMSC and ADSC. The research material was collected at several time intervals: before the administration of stem cells, after 24 and 72 h, and after 7 days. Blood samples were collected to assess hematological parameters and the level of pro-inflammatory cytokines, while the milk samples were used for microbiological assessment and to determine the somatic cells count (SCC). The administration of allogeneic MSCs resulted in a reduction in the total number of bacterial cells, Staphylococcus aureus, bacteria from the Enterobacteriaceae group, and a systematic decrease in SCC in milk. The therapeutic effect was achieved via intravenous + intramammary or intramammary administration.

Prevalence and distribution of transmissible venereal tumor in dogs from a rural community of Mexico

Objective: The aim of this work was to determine the rates of prevalence and recurrence of TVT infection in owned dogs, as well as the distribution of the disease, in Cuajinicuilapa, Guerrero, Mexico. Methods: This work was an observational, descriptive, longitudinal study. A TVT diagnosis was performed in all dogs in the community based on clinical records, a physical examination, and a cytological evaluation, and further confirmed by histopathology. Tissue samples were surgically obtained for histopathological analysis and, when required, to remove tumors and give treatment with vincristine; a GPS system was used to identify possible spatial groupings. All cases were re-evaluated one year after. Results: A prevalence of 5.15% was found in 1047 dogs (512 females and 535 males; 25 positive females and 29 positive males). TVT infection was more frequent in 2–3-years-old dogs. A spatial aggregation pattern was observed in the local cases. One year after, no lesions were found in the 29 animals treated. Conclusion: TVT is a common disease, and its distribution suggests its aggregation in certain areas of the locality. Further studies on the dynamics of dog populations in small cities are required.

Human Papillomavirus Genotype Detection and Cytology Using a New Self-Sampling Method.

High-risk human papillomavirus (hrHPV) testing using dry-type self-sampled vaginal specimens is becoming more widespread worldwide due to increased screening uptake. However, for the triage of hrHPV-positive women, a visit to a general practitioner is required for reflex cytology. This study aimed to evaluate the hrHPV detection capability of CellSoft®, a wet-type self-sampling method that also allows for cytology. Thirty-eight women aged 20 years and older were included in the study. The women self-sampled using CellSoft® after using an Evalyn® Brush. PCR-based HPV genotyping was performed on both specimens and hrHPV detection results of both devices were compared. Additionally, cytological exam was performed on CellSoft® samples. Overall agreement between self-sampling devices for the detection of hrHPV in CellSoft® and Evalyn Brush was observed in 97.4% (37/38) of participants. More hrHPV genotypes were detected with Evalyn Brush than with CellSoft®. Among the 22 CellSoft® hrHPV-positive cases, 11 (47.6%) were atypical squamous cells of undetermined significance or worse. CellSoft® hrHPV genotype detection results were in good agreement with those of Evalyn Brush. CellSoft® provided a sufficient cell volume for HPV testing and cytological evaluation.

Atypical glandular cells and predictive features of malignancy in Pap smears: A retrospective monocentric study.

The introduction of cytological screening with the Papanicolau smear significantly reduced cervical cancer mortality. However, Pap smear examination can be challenging, being based on the observer ability to decode different cytological and architectural features. This study aims to evaluate the malignancy rate of AGC (atypical glandular cells) category, investigating the relationships between cytological and histological diagnosis. Eighty-nine patients, diagnosed as AGC at cytological evaluation and followed up with biopsy or surgical procedure at Policlinico Gemelli Hospital, Rome, Italy, were included in the study. The cytopathological architectural (feathering, rosette formation, overlapping, loss of polarity, papillary formation, three-dimensional formation) and nuclear (N/C ratio, nuclear enlargement and hyperchromasia, mitoses, nuclei irregularity, evident nucleoli) features of AGC were evaluated. Statistical analyses were performed to assess cyto-histological correlation and determine the relevance of architectural and nuclear features in the diagnosis of malignancy. Of the 89 AGC patients, 48 cases (53.93%) were diagnosed as AGC-NOS and 41 (46.07%) were diagnosed as AGC-FN, according to the Bethesda classification system. The follow-up biopsies or surgical resections revealed malignancy in 46 patients (51.69%). The rates of malignancy for AGC-NOS and AGC-FN were 35.41% and 70.73% respectively. Furthermore, analysing cytopathological features, we found that both architectural and nuclear criteria were statistically significant (p < 0.05). Only overlapping, nuclear irregularity and increased N/C ratio were not found to be statistically significant for detecting malignancy. Cytological diagnosis of glandular lesions remains a valid tool, when appropriate clinical correlation and expert evaluation are available.

Diagnosing adult and pediatric extrapulmonary tuberculosis by MPT64 antigen detection with immunohistochemistry and immunocytochemistry using reproduced polyclonal antibodies.

Diagnosing extrapulmonary tuberculosis (EPTB) is challenging. Immunohistochemistry or immunocytochemistry has been used to diagnose tuberculosis (TB) by detection of MPT64 antigen from various extrapulmonary specimens and has shown good diagnostic performance in our previous studies. The test can distinguish between disease caused by Mycobacterium tuberculosis (Mtb) complex and nontuberculous mycobacteria and can be applied on formalin-fixed paraffin-embedded tissue. As the antibodies previously used were in limited supply, a new batch of polyclonal antibodies was developed for scale-up and evaluated for the first time in this study. Our aim was to assess the diagnostic accuracy of the MPT64 test with reproduced antibodies in the high burden settings of Pakistan and India. Patients were enrolled prospectively. Samples from suspected sites of infection were collected and subjected to histopathologic and/or cytologic evaluation, routine TB diagnostics, GeneXpert MTB/RIF (Xpert), and the MPT64 antigen detection test. Patients were followed until the end of treatment. Based on a composite reference standard (CRS), 556 patients were categorized as TB cases and 175 as non-TB cases. The MPT64 test performed well on biopsies with a sensitivity and specificity of 94% and 75%, respectively, against a CRS. For cytology samples, the sensitivity was low (36%), whereas the specificity was 81%. Overall, the MPT64 test showed higher sensitivity (73%) than Xpert (38%) and Mtb culture (33%). The test performed equally well in adults and children. We found an additive diagnostic value of the MPT64 test in conjunction with histology and molecular tests, increasing the yield for EPTB. In conclusion, immunochemical staining with MPT64 antibodies improves the diagnosis of EPTB in high burden settings and could be a valuable addition to routine diagnostics.

Veterinary clinicians prefer template-style reports with personal confidence estimates for cytologic sample evaluations.

To examine preferences of veterinary clinical pathologists, clinicians, and students for cytology report formats. 24 clinical pathologists, 1,014 veterinarians, and 93 veterinary students who were members of the Veterinary Information Network. Members of the Veterinary Information Network responded to an online survey invitation, made available between July 11, 2023, and July 24, 2023. Respondents were randomly directed to 1 of 4 sets of cytology reports, each containing a traditional narrative format, narrative format with terms expressing a degree of confidence and associated numerical ranges, and template format with similar estimates of confidence. Respondents ranked the reports in order of preference and then provided comments about their top-ranked choice. Responses were analyzed mostly with descriptive statistics or comparisons of proportions. 14 of 24 clinical pathologists preferred the traditional narrative format, whereas 449 of 1,042 veterinary clinicians and veterinary students preferred the template format. Respondents (460/1,131) ranked the template format as most preferred, but the narrative format with terms expressing a degree of confidence ranked highest overall. Many respondents appeared to misunderstand the degree of confidence estimates being expressed numerically. Respondents choosing each format often stated that their preferred choice was "easiest to understand" and "most comprehensive." Given the preferences of veterinary clinicians and veterinary students for a template format, clinical pathologists should consider modifying the way they report evaluations of cytologic specimens. Template formats should help standardize reporting of cytologic specimens, thereby improving communication between clinical pathologists and clinicians. However, both clinicians and clinical pathologists need to better understand the purpose of terminology expressing degrees of confidence in such reports.

3D Bioprinting-Based Dopamine-Coupled Flexible Material for Nasal Cartilage Repair.

Since 3D printing can be used to design implants according to the specific conditions of patients, it has become an emerging technology in tissue engineering and regenerative medicine. How to improve the mechanical, elastic and adhesion properties of 3D-printed photocrosslinked hydrogels is the focus of cartilage tissue repair and reconstruction research. We established a strategy for toughening hydrogels by mixing GelMA-DOPA (GD), which is prepared by coupling dopamine (DA) with GelMA, with HAMA, bacterial cellulose (BC) to produce composite hydrogels (HB-GD). HB-GD hydrogel scaffolds were characterized in vitro by scanning electron microscopy (SEM), Young's modulus, swelling property and rheological property tests. And biocompatibility and chondrogenic ability were tested by live/dead staining, DNA quantitative analysis and immunofluorescence staining. Combined with 3D bioprinting technology, mouse chondrocytes (ADTC5) were added to form a biological chain to construct an in vitro model, and the feasibility of the model for nasal cartilage regeneration was verified by cytology evaluation. With the increase of GD concentration, the toughness of the composite hydrogel increased (47.0 ± 2.7 kPa (HB-5GD)-158 ± 3.2 kPa (HB-20GD)), and it had excellent swelling properties, rheological properties and printing properties. The HB-GD composite hydrogel promoted the proliferation and differentiation of ATDC5. Cells in 3D printed scaffolds had higher survival rates (> 95%) and better protein expression than the encapsulated cultures. The HB-10GD hydrogel can be made into a porous scaffold with precise shape, good internal pore structure, high mechanical strength and good swelling rate through extrusion 3D printing. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Abstract 1273: Tumor progression and tumor microenvironment of pancreas cancer ascites revealed by scRNA-seq and spatial transcriptomics

Abstract Peritoneal metastasis of pancreas cancer is one of the ultimate modes of spread in the cancer. Little is known about how cancer cells spread into abdominal cavity and what is different from hematogenous metastasis. It is also not well characterized how tumor microenvironment prevent cancer cells from spreading in abdominal cavity. We performed single-cell RNA-seq analysis and special transcriptomic analysis to clarify the progression and environment of the cancer. We collected and processed ascites samples of 20 PDAC samples from 19 patients. Sixteen ascites samples were obtained from abdominal paracentesis and two cases in the postmortem period. For one patient ascites was collected both pre and postmortem. All but one patient received chemotherapy prior to ascites collection. Routine clinical cytology evaluation of ascites samples from 17 patients indicated it was malignant in 11 patients and nonmalignant in six patients. All ascites samples were purified by gradient centrifuge using Ficoll-Paque Plus to remove erythrocytes and necrotic debris. For the platform, we adopted the 10x Genomics Chromium Single Cell Gene Expression platform for single-cell RNA-seq library preparation. Spatial transcriptomics was performed for eight FFPE samples from the two autopsy patients with paired cancer ascites using 10x Visium. The CytAssist was used to transfer transcripts of pre-sectioned and pre-stained tissues with 11 × 11 mm Visium Capture Area slides. The sample locations were metastatic tumors to bowel wall, rib bone, mesentery, iliac nerve, diaphragm, peritoneum, pelvis, and primary pancreas tumor. A total of 248,263 cells were analyzed in single-cell RNA-seq. Cell population in pancreas cancer ascites consisted of macrophages, lymphocytes, dendritic cells, plasma cells, plasmacytoid dendritic cells, mesothelial cells, and cancer cells. Cancer cells were detected in twelve samples whereas no cancer cells were detected in eight samples. Gene ontology and gene set enrichment analyses showed macrophages in ascites with cancer cells involved neutrophil activation while those in ascites without cancer cells retained antigen presentation capacity and inducing interferon gamma pathway. Mesothelial cells in ascites with cancer cells showed more apoptosis and lymphocyte proliferation while those in ascites without cancers cells triggered fibrosis and mitosis. These data suggest that macrophage and mesothelial cells could be categorized into anti-tumor cells and tumor associated cells. Ongoing investigations are exploring ligand-receptor relationships. We are going to compare the expression profile of tumor cells in ascites fluid and in tissue from various organs by integrating single cell RNA-seq data and special transcriptomics data. Citation Format: Shigeaki Umeda, Elias-Ramzey Karnoub, Fiyinfolu Balogun, Alejandro Jiménez-Sánchez, Christine A. Iacobuzio-Donahue. Tumor progression and tumor microenvironment of pancreas cancer ascites revealed by scRNA-seq and spatial transcriptomics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1273.

Diagnostic Efficacy of a Novel Rotating Brush for Endoscopic Sampling of Malignant Biliary Strictures: A Multicenter Prospective Study.

Although cytologic examination of biliary stricture brushings obtained by endoscopic retrograde cholangiopancreatography is commonly used for diagnosing malignant biliary strictures (MBSs), it has low sensitivity. Several new brushes have capabilities that are still being debated. We have developed a novel brush working from conventional back-and-forth movement to rotation in situ (RIS) that may be more efficient for MBS sampling. We aimed to compare the MBS detection sensitivity of our RIS brush with that of the conventional brush. In this multicenter prospective study, we enrolled patients who underwent endoscopic retrograde cholangiopancreatography for suspected MBSs involving biliary stricture brushings obtained using our RIS brush. The historical control group consisted of the 30-brushing arm of our previous randomized trial (patient inclusion, 2018-2020) that used the study design in the same centers and with the same endoscopists as were used in this study. The primary outcome was to compare the sensitivity and specificity of detecting MBSs by cytologic evaluation of biliary stricture brushings between the 2 groups. We enrolled 155 patients in the intent-to-treat analysis. Using the same number of brushing cycles, the RIS brush showed a higher sensitivity than the conventional brush (0.73 vs 0.56, P = 0.003). In per-protocol population, the sensitivity was also higher in the RIS brush group than in the conventional brush group (0.75 vs 0.57, P = 0.002). Multivariate analysis revealed that the RIS brush was the only predictive factor for MBS detection. No significant differences were observed in procedure-related complications between the 2 groups. The RIS brush was a promising tool for effective and safe MBS sampling and diagnosis. Further randomized studies are warranted to confirm our results (Chictr.org.cn, identifier: ChiCTR2100047270).

Endobronchial ultrasound-transbronchial needle aspiration: effectiveness and accuracy in non-small cell lung cancer staging.

Endobronchial ultrasound-transbronchial needle aspiration (EBUS-TBNA) has a cardinal role in the diagnosis and staging of non-small cell lung cancer (NSCLC), providing an accurate nodal staging in a less invasive way than surgical biopsy. The aim of this study was to assess the diagnostic accuracy of EBUS-TBNA in the pre-operative NSCLC mediastinal staging, as well as to evaluate EBUS-TBNA specificity and sensibility in our cohort. We retrospectively analyzed data of NSCLC patients who underwent EBUS-TBNA followed by major pulmonary resection between January 2020 and December 2022. EBUS-TBNA was performed in patients with NSCLC (central T ≤ 3cm, peripheral/central T > 3cm), following the ESTS guidelines. The target nodes were selected on the basis of their radiologic/metabolic characteristics. Each procedure was conducted together with rapid on-site cytological evaluation (ROSE). Twenty-five patients were included (M/F = 17/8). At least three needle passages on each target lymph node were performed. No complications during or after the procedures occurred. We found a 100% correspondence between ROSE on the sampled nodes and postoperative pathologic findings. An upstaging occurred in three cases (12%) because of the involvement of stations 5 and 6 (not accessible via EBUS), while the only case of downstaging (N2 → N0, 4%) was probably due to intercurrent neoadjuvant chemotherapy. In all cases, EBUS-TBNA has proved to achieve a diagnostic procedure on the target nodes. EBUS-TBNA is a safe and effective procedure that offers high sensitivity and specificity when performed together with ROSE, which improves the accuracy of sampling. Doubt on nodal stations 5 and 6 involvement should be settled by other techniques.

A case of lung FNA with atypical squamous cells.

Fine-needle aspiration (FNA) is a commonly employed method for initial diagnosis and work-up of pulmonary nodules. Utilization of rapid on-site evaluation (ROSE) has the added benefit of allowing for triaging of material as appropriate for ancillary studies including microbiology testing. While many pulmonary lesions are easily categorized by cytologic evaluation, more difficult cases exist. In particular, pulmonary lesions demonstrating atypical squamoid cells can cause diagnostic challenge given the morphologic overlap between benign and malignant pulmonary lesions showing atypical squamoid cells. We herein review these common and uncommon squamoid lesions, which may enter within the differential when encountering atypical squamoid cells in pulmonary FNA specimens with emphasis on morphologic pitfalls and approaches to appropriate categorization.

Diagnostic utility of intraoperative squash smear cytology of Intra-cranial lesions in a resource limited setup of central India

IntroductionIntraoperative squash smear cytology is a useful diagnostic tool in case of CNS lesions. In resource poor countries where frozen section is unavailable, cytology is the mainstay method in giving a rapid intraoperative diagnosis which helps Surgeons regarding the extent of excision. The current study aims to assess the feasibility and accuracy of intraoperative squash cytology in evaluation of CNS tumors. Definitive diagnosis is confirmed by histopathological examination. Materials and methodsThis retrospective observational study was carried out at Department of Neuropathology. All patients diagnosed with space occupying lesion (SOL) in CNS were enrolled in the study. Intra-operative cytological diagnosis was compared to histopathological diagnosis. WHO classification of CNS tumors 2021 was used to classify tumors. ResultsTotal seventy patients with intracranial SOLs were enrolled in this study who were operated for the same and their specimens were examined. Two samples were found to be inadequate and not included in final analysis. Mean age was 38.8 ± 17.85 (SD). Male–female ratio was 1.5:1 with 41 (60.3%) males and 27 (39.7%) females. 58 had neoplastic and rest 10 had non-neoplastic lesions. Complete concordance was achieved in 61 cases (Diagnostic Accuracy-88.2%). 3 (4.4%) cases showed partial concordance and 4 (5.9%) were discordant. For detection of malignant lesions through squash smear cytology overall Sensitivity was 73.9%, Specificity-97.8%, Positive Predictive Value-94.4% and Negative Predictive Value-88%. ConclusionsSquash smear cytology is a simple, rapid and cost-effective method relying solely on the expertise of pathologist. High diagnostic accuracy can be achieved with squash cytology by taking clinical and radiological findings into consideration.

Impression Cytologic Evaluation of the Conjunctiva in Patients Treated with Topical 1% Voriconazole

The aim of the present study was to evaluate any conjunctival metaplastic changes by impression cytology in patients who underwent topical 1% voriconazole treatment for severe fungal keratitis. The study was conducted at Ege University Faculty of Medicine, Departments of Ophthalmology and Medical Pathology. Patients who were treated with 1% topical voriconazole for fungal keratitis for at least 3 months were included. The used topical voriconazole treatment was initiated as one drop every hour and was tapered according to clinical improvement in all patients. Treatment was continued 4 times a day for at least 3 months. Impression cytology samples were collected at least 3 months after cessation of topical voriconazole from the affected eyes and from the fellow eyes as a control group. Collected specimens were transferred to the pathology department for evaluation and grading (Nelson’s grading system). The mean age of the patients was 57.68±17.32 years (range, 22-87 years). The impression cytology grade of the inferior bulbar conjunctiva was 1.73±0.77 (range, 0-3) in the study group and 1.19±0.98 (range, 0-3) in the control group (p=0.03). The impression cytology grade of the temporal bulbar conjunctiva was 1.69±0.73 (range, 0-3) in the study group and 1.15±0.88 (range, 0-3) in the control group (p=0.02). The impression cytology grades of the nasal and superior bulbar conjunctiva did not differ statistically (p values 0.13 and 0.17, respectively). Topical voriconazole is an effective broad-spectrum antifungal drug, but it induces conjunctival squamous metaplasia. Clinicians should be aware of this possible side effect of topical voriconazole and should carefully evaluate the conjunctiva of treated patients at each visit to detect possible metaplastic changes.