Journal articles on the topic 'Cytology; AgNOR count and Effusions'

We compared the performance of clinicopathologic and molecular tests used in the antemortem diagnosis of feline infectious peritonitis (FIP). From 16 FIP and 14 non-FIP cats, we evaluated retrospectively the sensitivity, specificity, and likelihood ratios (LRs) of serum protein electrophoresis, α1-acid glycoprotein (AGP) on peripheral blood, screening reverse-transcription nested PCR (RT-nPCR) on the 3’–untranslated region (3’-UTR), and spike (S) gene sequencing on peripheral blood, body cavity effusions, and tissue, as well as body cavity cytology and delta total nucleated cell count (ΔTNC). Any of these tests on blood, and especially the molecular tests, may support or confirm a clinical diagnosis of FIP. A negative result does not exclude the disease except for AGP. Cytology, 3’-UTR PCR, and ΔTNC may confirm a clinical diagnosis on effusions; cytology or 3’-UTR PCR may exclude FIP. Conversely, S gene sequencing is not recommended based on the LRs. On tissues, S gene sequencing is preferable when histology is highly consistent with FIP, and 3’-UTR PCR when FIP is unlikely. Combining one test with high LR+ with one with low LR− (e.g., molecular tests and AGP on blood, ΔTNC and cytology in effusions) may improve the diagnostic power of the most used laboratory tests.

Background: Exudative pleural effusions are a common diagnostic problem in clinical practice, as the list of causes is quite exhaustive, although sometimes they can be inferred from the clinical picture. In the West the most common cause is Para pneumonic effusions followed by malignancy, while in India it is tubercular effusion followed by malignant effusion. Despite the availability of various tests, there is a need for defining the best diagnostic and cost-effective approach to quickly diagnose and treat exudative pleural effusions. The objectives are to conduct a clinical and etiological study of exudative pleural effusion, to evaluate biochemical profile, cytological profile and radiological profiles of exudative pleural effusion.Methods: Prospective study of 100 patients with exudative pleural effusions. The demographic data was expressed as mean±standard deviation. Comparison between groups was done by Chi-Square test and Fischer exact test for categorical variables and Kruskar-Wallis and Mann-Whitney tests for continuous variables.Results: There were 67 males and 33 females. The mean age was 41.6±15.74. The majority were tubercular in origin (67%),13%,8%,3%and 6% were malignant effusions, Synpneumonic effusion, pancreatic effusions and empyema respectively. Diagnosis was not established in 3% of effusions. Massive effusions were seen in 53.8% of malignant effusions and 33.3% of empyemas. Most effusions had a total cell count between 1000 to 5000 cells /mm3.Lymphocyte predominant effusions were seen in 84.6% and 89.6% of malignant and tubercular effusions. 61.5% of malignant effusions had a positive cytology. Tubercular effusion had a pleural fluid ADA more than 40 IU/L. 92.3% of malignant effusion had pleural fluid ADA less than 30IU.Conclusions: Pleural effusion is a commonly encountered in medical practice and in our country, the commonest cause is tuberculosis, as is evidenced from the present study. The initial step in evaluating case of pleural effusion is to establish the cause of pleural effusion which is done by a detailed history, clinical examination and investigations like a chest radiology and pleural fluid analysis. Even in the advanced diagnostic approaches, still detailed clinical history and examination of the patient of the patient is important to make a clinical diagnosis. All suspected cases of pleural effusion should undergo Sonography of the thorax along with routine chest x-ray. Fluid cytology should be done to confirm tuberculosis or to rule out malignancy, which guides the physician for further evaluation of the patient if required.

The cell count differential of pleural fluid sample is of great importance for estimation of the nature of pleural effusion. In the present article, we compared the efficiencies of routine cytology method with light microscopy, cytological examination with centrifuge Cytospin-4 and immunocytochemical methods. We have studied cytological samples from 1597 patients, with pleural effusion. Effusions associated with malignancies were reported in 22.7 % of patients including carcinomatosis (74.6 %), primary tumors of pleura (21.5 %), effusions associated with non epithelial malignancies (3.9 %). Benign pleural effusions were reactive (63.6 %), tuberculotic (13.5 %), "cholesterol pleurisy" and chylothorax (0.2 %). Carcinomatous pleuritis was found in patients with lung carcinoma (55.4 %), breast cancer (21.8 %) and ovary cancer (12.2 %). Specific malignant features (direct and indirect) were noted in pleural fluid on breast cancer, carcinomas of ovary, stomach, kidney, small cell lung carcinoma and squamous cell lung carcinoma. These features are hardly detected in patients with malignancies of intestines, prostate and endometria because these types of tumours are rarely metastatic to pleura. We were failed to define particular features of lung adenocarcinoma. The centrifuge Cytospin-4 was used in the most difficult cases (13.5 %) providing minimal number of presumable diagnosis. Primary tumours of pleura are the most difficult for detection. Immunocytochemical analysis found monoclonal mesothelial cell of НВЕМ 1 clone, cytokeratin, vimentin to be positive and carcinoembry onic antigen, Ber-EP4, CD-15 to be negative in the studied tumors.

<strong>Introduction:</strong>&nbsp;Ascitic fluid cytology is the most efficient as well as reliable mode for the diagnosis of cause of ascites. Malignancy could be ruled out by ascitic fluid cytology with consideration to the clinical presentation and previous history. This study is conducted aiming to study cytology of ascetic fluid in various diseases to establish clinic-cytological correlation for better management of patients.&nbsp;<strong>Material and Methods:</strong>&nbsp;A total of 115 samples were collected and was subjected to various physical as well as biochemical and cytological examination done in the department of pathology.&nbsp;<strong>Result:</strong>&nbsp;From the samples collected and analysed, 81.74% were cytologically non malignat and the remaining 18.26% were malignant. Moreover 62.61% were females and 37.39% were males. Transudative and exudative effusion accounted for 66.95% and 33.04% respectively. Most of the malignant effusions were exudative.&nbsp;<strong>Conclusion:&nbsp;</strong>This study concludes that most useful tests used in establishing the diagnosis of peritoneal effusion are peritoneal fluid cytology and peritoneal fluid cell count. It is a complete diagnostic modality which aims at pointing out the etiology of effusion as well as in certain cases a means of prognostication of the disease process. Non-malignant causes are more common causes of peritoneal effusion.

The objective of this study was to evaluate the cellular proliferative potential of oral lichen planus (OLP) lesions from patients without hepatitis C virus (HCV) by means of AgNOR method, as well as the cellular proliferative potential of the normal oral mucosa from patients with HCV, treated or untreated by interferon and ribavirin. A cross-sectional study was developed to investigate four groups: 10 HCV+ patients without clinical signs of OLP who had never been treated for HCV infection - Group 1; 10 HCV+ patients that were under interferon and ribavirin treatment - Group 2; 15 patients with reticular OLP lesions histopathologically confirmed, without HCV - Group 3; and 15 blood donors without HCV infection and no clinical signs of OLP GROUP 4 Control Group. The cytological material of all groups was collected by the liquid-based cytology technique. Then, the sedimented material from each patient was filled with the Nucleolar Organizer Regions impregnation by silver method (AgNOR). The count of NORs was performed on 100 epithelial cell nuclei per patient using the Image Tool(tm) software. The Tukey HSD test was used to compare the median value of NORs among the groups and showed that the oral mucosa of HCV+ patients previously treated with anti-HCV drugs (GROUP 2), presented a higher average number of NORs in relation to others (p&lt;0.05). The anti-HCV treatment may be related to increased cell proliferation of oral mucosa, indicating a possible relationship between OLP and HCV+ patients treated with interferon and ribavirin.

Cases involving pleural and peritoneal effusions occur relatively frequently in clinical practice. Determining the underlying etiology in these cases relies mainly on fluid analysis. The technique used for obtaining the pleural or peritoneal fluid can impact greatly the results of the analysis. Most often used diagnostic tools include evaluation of gross appearance, Total Nucleated Cell Count / Total Protein (TNCC/TP) measurement, chemical/biochemical analysis (Lactate dehydrogenase and lactate, cholesterol, triglycerides, glucose, creatinine, pH, pO2, pCO2, and K measurements), cytology (identification of septic and non-septic inflammation and neoplasia), microbiology (Gram stain, culture, molecular techniques), and specific tests for certain clinical conditions and diseases. Classifying an effusion as transudate, modified transudate and exudate is traditionally based on the TNCC and TP values. New diagnostic methods encourage the clinician to approach the effusion etiologically instead of strictly following this traditional classification. Many of the diagnostic tests described in this review are simple and can be performed in-house, providing the clinician quickly with information about the cause of the effusion, essential for an effective treatment plan without wasting valuable time.

Malignant pleural effusion (MPE) is defined as an effusion that occurs in association with malignancy, as evidenced by the finding of malignant cells on pleural fluid cytology or pleural biopsy. The pathophysiology of MPE is not yet clear, but several hypotheses have been developed to explain the mechanism of MPE. Accumulation of effusion in the pleural cavity occurs due to increased vascular permeability due to the inflammatory reaction caused by the infiltration of cancer cells in the parietal and/or visceral pleura. Other possible mechanisms are the direct invasion of the tumor adjacent to the pleura, obstruction of the lymph nodes, hematogenous spread, and primary pleural tumor. Pleural fluid from a malignant process is usually an exudate. To distinguish between exudate and transudate is to assess the protein and LDH levels of the pleural fluid. Hemorrhagic pleural fluid with a red blood cell count &gt;100,000/mm3 suggests an MPE. Pleural fluid glucose levels at low &lt; 60 mg/dl at about 15-20% MPE. The definitive diagnosis of MPE is by finding malignant cells in the pleural fluid (cytology) or pleural tissue (pathological histology). Management of malignant pleural effusions is principally palliative. Management that is often done in cases of MPE is therapeutic thoracentesis, pleurodesis, drainage with long-term indwelling catheter, manufacture of the pleuroperitoneal shunt, intrapleural therapy, and radiotherapy.

Chylothorax is a rare disease (1-2% of pleural effusions), with a prevalence between 1/8600 and 1/15,000 births. It is characterized by the presence of chyle in the pleural cavity. Three categories of chylothorax are known: congenital, idiopathic or traumatic (usually postoperative) chylothorax. We report the observation of a 3-year-old child with idiopathic chylothorax revealed by respiratory symptoms, with pleural effusion and ipsilateral lung collapse on chest x-ray and CT. Cytology and chemical analysis of pleural fluid showed exudative fluid with a chyous appearance, high cell count (2800 cells/mm3), lymphocyte predominance (98%), and culture was sterile. Chylothorax is usually revealed by dyspnea, but also by nausea, vomiting, anorexia and/ or malnutrition. The diagnosis is suspected when the puncture brings back a milky liquid, confirmed by the presence of a triglyceride level greater than 1.2 mmol/ L and number of cells greater than 1000 cells/ mL, with lymphocyte predominance. Treatment of chylothorax can be conservative or surgical when conservative management fails and aims to stop permanent chyle leakage.

Abstract Abstract 4834 Multicentric Castleman's Disease(MCD) can have an aggressive course making early diagnosis and treatment important. Although rare, peripancreatic involvement is known in Castleman's disease and an unusual presentation should be considered especially in the setting of HIV. An excisional lymph node biopsy is required for diagnosis. High plasma titers of Human Herpes Virus 8 DNA should also raise this possibility. Coexistence of Castleman disease and body cavity lymphoma has been reported but is rare. Case Presentation: A 42 year old African American gentleman with a history of HIV and CD4 count 271 presented with intractable nausea and vomiting of 1 month duration; associated with epigastric pain and weight loss. Past history was significant for presentation with jaundice 6 months earlier when he was found to have a pancreatic head mass with portocaval and retroperitoneal lymphadenopathy raising the suspicion of pancreatic adenocarcinoma. Endoscopic ultrasound guided fine needle aspiration of the mass had revealed a poorly differentiated malignant neoplasm that could not be further characterized by immunohistochemistry. Considering the HIV status of the patient, there was a strong suspicion of lymphoma. A PET-CT was planned to identify an appropriate site for tissue diagnosis but the patient was lost to follow up. Repeat imaging during the current admission revealed increasing size of the pancreatic mass causing complete obstruction of second part of duodenum. The patient subsequently underwent gastro-jejunostomy to bypass the duodenal obstruction and an excisional biopsy of two mesenteric lymph nodes was performed during the procedure. Pathology revealed HHV-8 positive plasma cell variant of Castleman's disease. Patient also had high plasma titers of HHV-8. Furthermore, the patient was also found to have bilateral pleural effusions and a thoracentesis was performed. Pleural cytology revealed large plasmablastic lymphoid cells that were positive for HHV-8, CD30, CD45, CD138, Ebstein-Barr virus (EBV), epithelial membrane antigen, and multiple myeloma oncogene-1. These findings suggested a diagnosis of body cavity lymphoma. Discussion: Although rare, the association of Multicentric Castleman's disease (MCD) with HIV is well recognized. The incidence is independent of CD4 count and the use of HAART. There are isolated case reports of pancreatic involvement with Castleman's disease and only one involving the duodenum. While most patients with peripancreatic involvement presented with abdominal discomfort, nausea or systemic symptoms, our patient was severely symptomatic from complete duodenal obstruction. Our case also highlights that the diagnosis can be elusive and requires excisional lymph node biopsy. Moreover, HIV MCD is fluorodeoxyglucose avid, and a PET-CT may help identify the gland to biopsy next in difficult to diagnose cases. Body cavity lymphoma is also HHV-8 related and this diagnosis should be considered in any HIV patient with effusions. Pleural fluid cytology and immunohistochemistry usually clinch the diagnosis. Disclosures: No relevant conflicts of interest to declare.

Abstract Abstract 4556 A 37 year old male with T cell acute lymphoblastic leukemia with normal cytogenetics 46XY in second complete remission (CR2) after 8 cycles of HyperCVAD and 4 cycles of salvage nelarabine underwent myeloablative conditioning with cyclophosphamide TBI for a matched related donor allogeneic stem cell transplant from his brother. His day+100 bone marrow biopsy showed CR2 and was 100% donor. His post transplant course was unremarkable until he relapsed d+162. The immunosuppression was tapered off rapidly and 2 cycles of nelarabine and a donor lymphocyte infusion (1×107 CD3 cells/kg) were completed by d+254 and CR3 was achieved. He did well until d+608 when he presented with a month long history of dyspnea and recurrent bilateral large pleural effusions (despite multiple large volume thoracenteses) and anasarca including facial edema. He had gained 15 kilograms in 2 weeks. He had no fever, skin rash, sicca syndrome or lichen planus. Chest and abdomen CT showed massive bilateral pleural effusions, but no lung infiltrate, pericardial effusion or ascites or lymphadenopathy. The complete blood count, renal function and liver function tests were normal and there was normal hepatopedal flow in the main portal vein and its branches. The echocardiogram showed a left ventricular ejection fraction of 60% (normal) and there were no vegetations. The serum ANA, anti-Ro and anti-La were negative. The bone marrow biopsy showed no evidence of relapse of T cell ALL. The peripheral blood STR was 100% donor. The pleural fluid was transudative and cytology showed no malignancy. Pleural fluid flow cytometry showed no CD20+ B cells but did show polyclonal T cells with a CD4:CD8 ratio of 0.22. The pleural biopsy showed reactive mesothelial cells with mixed acute and chronic inflammation and no malignancy. Ebstein Barr virus in-situ hybridization and HHV-8 immunohistochemistry was negative on the pleural biopsy. All bacterial, fungal and viral (including CMV, EBV, HSV, adenovirus, VZV, HIV, BK) of the blood, urine and pleural fluid were unremarkable. A random skin biopsy of the forearm showed scleroderma. Methylprednisolone 2 mg/kg iv daily and therapeutic tacrolimus with extracorporeal photoporesis was begun with resolution of the pleural effusions and anasarca two months later. On d+903 he remains in CR3 and is on tapering courses of prednisone and tacrolimus without recurrence of the pleural effusions or anasarca. Conclusion: Isolated sterile pleural effusions with anasarca should be recognized as a rare feature of late onset chronic graft versus host disease so as to allow prompt initiation of immunosuppression. The patient shared the HLA antigens HLA DR15 and HLA DQW6 which are associated with autoimmune disease such as systemic lupus erythematosus suggesting that patients with this HLA combination may be more prone to this presentation. Disclosures: No relevant conflicts of interest to declare.

Objective: To determine the etiology and outcomes of pleural effusions in children. Study Design: Cross-sectional study. Setting: Department of Pediatric Medicine, National Institute of Child Health, Karachi, Pakistan. Period: June 2022 to October 2023. Methods: A total of 60 admitted children of either gender, aged 1-12 years, and having pleural effusion were analyzed. Relevant laboratory and radiological studies including complete blood count, complete urinalysis, ultrasonography, chest X-ray, sputum for acid fast bacilli (AFB) smear, and gene x-pert were performed. Pleural tap was done and fluid was sent to institutional laboratory for biochemical analysis, microbiological testing, and cytology testing to rule out infections. Results: In a total of 60 children, 34 (56.7%) were male. The mean age was 5.4±3.5 years. Cough and fever were the most reported symptoms among the patients, with 18 (30.0%) having a cough, and 17 (28.3%) having fever. The sputum test was positive in 7 (11.7%) of children. The cause of pleural effusion in 44 (73.3%) of patients was pneumonia, 12 (20.0%) tuberculosis, and 4 (5.7%) had congestive heart failure (CHF). Conclusion: The cause of pleural effusion in the pediatric age group is commonly infection. Taking preventive measures, diagnosing and managing pleural effusion promptly as a multidisciplinary approach can decrease the morbidity and mortality rate.

BACKGROUND The diagnostic yield of thoracoscopy is 95 %, of pleural fluid cytology it is 62 % and of closed pleural biopsy is 44 %, in malignant effusion. We wanted to study the diagnostic utility of flexible thoracoscopy in undiagnosed exudative pleural effusion and compare the thoracoscopy findings with the histopathology results. METHODS The study was conducted in the Department of Respiratory Medicine, Government Stanley Medical College, Chennai, from January 2019 to January 2020. 40 patients were enrolled in this longitudinal observational study with moderate to massive effusion and were evaluated with pleural fluid aspiration and sent for cytology, protein sugar analysis, total count, and ADA. Those cases which are exudative pleural effusions, with ADA value of less than 40 IU / L were subjected to thoracoscopy after being evaluated for fitness for thoracoscopy with complete blood count, bleeding time, clotting time, sputum for AFB, ECG, pulse oximetry, cardiac evaluation and CT chest. RESULTS Thoracoscopy was done in 40 enrolled patients. In this study, biopsy was taken from the parietal pleura in all the cases. Of these 40 cases, 30 were male and 10 were female, that is 75 % males and 25 % females. The mean age of the study population was 43 ± 14.9. Patient with the lowest age in this study group was 18 years and highest was 71 years. 16 cases (40 %) presented with left sided pleural effusion. 24 cases (60 %) presented with right sided pleural effusion. 30 cases presented with massive effusion, and 10 cases with moderate effusion. Of the 40 cases, 27 cases presented with straw coloured pleural effusion. 13 cases were haemorrhagic effusion. Histopathologic examination showed 11 cases as malignant and 29 cases as non-malignant out of which 18 cases were of tuberculosis aetiology. Thoracoscopy revealed adhesions in 13 cases and mass lesion in 4 cases. Of the 4 mass lesions 3 came as malignant, normal pleura in 11 cases, 10 were non-malignant and 1 was malignant. Nodules were seen in 12 cases of which 7 came as malignant. Straw coloured effusion was seen in 27 cases, of which 2 were malignant. CONCLUSIONS The most important indication for thoracoscopy is exudative undiagnosed pleural effusion. The overall diagnostic yield in pleural fluid cytology is 62 % and blind pleural biopsy is 44 %. The diagnostic yield of thoracoscopy varies from 60 % to 97 % in various studies, whereas, in our study, it is 72.5 %. Visualization of the visceral and parietal pleura is another advantage, so that we can take biopsy from the abnormal areas. KEYWORDS Flexible Thoracoscopy, Undiagnosed Exudative Pleural Effusion

Aspiration of serous cavities is a simple and relatively non-invasive technique to achieve a diagnosis. The information provided by body fluid analysis serves several functions; it is a complete diagnostic modality which aims at pointing out the etiology of effusion as well as in certain cases, a means of prognostication of the disease process. The diagnostic performance of the cytological study of the fluid may be attributable to the fact that the cell population present in sediment. Thus, this study was aimed to assess the role of pleural fluid analysis in establishing the cause of pleural effusion. A retrospective study was carried out in the Department of Pathology, College of Medical Sciences Teaching Hospital, Bharatpur from July 2008 to July 2010. Two hundred samples of pleural fluid were examined for total cell count, cell type and cellular features along with biochemical study to find out the level of protein, glucose and chloride. In two hundred samples, 80% were exudative and 20% were transudative. Cirrhosis and congestive cardiac failure were commonest causative factors in transudatives. The cases of Tuberculosis were predominant in exudatives followed by malignancies. In the samples from all the tuberculosis patients, lymphocytes were the predominant cell type. Most of malignant effusions were exudatives. The primary site could be assessed by cytological examination in more than fifty percent of cases. The present study demonstrates that the most useful test in establishing the diagnosis of pleural effusion is pleural fluid analysis which includes mainly its cytology and cell count. Journal of College of Medical Sciences-Nepal,2011,Vol-6,No-4, 36-45 DOI: http://dx.doi.org/10.3126/jcmsn.v6i4.6724

Pleural effusion is the accumulation of fluid between the parietal and visceral pleura called the pleural cavity. It can occur by itself or can be the result of surrounding parenchymal diseases like infection, malignancy or inflammatory conditions. Pleural effusion is one of the major causes of pulmonary mortality and morbidity. Both the visceral and the parietal pleura play an important role in fluid homeostasis in the pleural space. Pleural effusions develop when there is excess hydrostatic pressure in the pulmonary capillaries, when fluid removal is impaired by compromised lymphatic drainage or when protein and cell rich fluid enters the pleural space through leaky capillary and pleural membranes. Pleural fluid is classified as a transudate or exudate based on modified Light’s criteria, proposed by Light et al., in 1972 which has been the standard differentiation method. It is considered an exudative effusion if at least one of the criteria is met:  Pleural fluid protein/serum protein ratio of more than 0.5  Pleural fluid lactate dehydrogenase (LDH)/serum LDH ratio of more than 0.6  Pleural fluid LDH is more than two-thirds of the upper limits of normal laboratory value for serum LDH. Commonly performed tests on the pleural fluid to determine etiology are a measurement of fluid pH, fluid protein, albumin and LDH, fluid glucose, fluid triglyceride, fluid cell count differential, fluid gram stain and culture and fluid cytology.

Objectives: Pleural fluid evaluation is an effective modality for identifying actionable genetic mutations to guide therapy in lung carcinoma. Clinicians requesting molecular studies often send large volumes of fluid to be processed that is not possible or cost effective and is hence not standard of practice in most cytopathology laboratories. We wanted to establish the characteristics of an adequate specimen that would yield reliable results with current molecular testing platforms. Material and Methods: A review of 500 malignant pleural effusions, from pulmonary and non-pulmonary sources, was undertaken over a 4-year period. Of these 44 cases (from 42 patients) that were positive for primary lung adenocarcinoma were included in the study. Molecular analysis was performed on 42 specimens. A complete next generation sequencing (NGS) panel was performed on 36 specimens. Individual testing for estimated glomerular filtration rate, KRAS, anaplastic lymphoma kinase, and ROS1 was performed on six specimens. The number of malignant cells and proportion of tumor to non-tumor nucleated cells (T: NT) on cell blocks was recorded as &lt;20%, 20–50% and &gt;50%. Results: The minimum volume on which a complete NGS panel could be performed was 20 ml with cell count of 1000 and T: NT proportion of 20–50%. The minimum number of tumor cells required for successful molecular analysis for T: NT proportion of &lt;20%, 20–50%, and &gt;50% was 300, 250, and 170 cells, respectively. Conclusion: We concluded that tumor cell proportion, rather than specimen volume, is of prime importance for determining the efficacy of pleural fluid for molecular studies. Evaluation of both absolute and relative numbers of tumor cells is critical for assessing the adequacy and predicting successful yield for molecular analysis.

Background: Pleural effusion is a common finding in patients. For a long time, a light criterion is used to analysis of pleural effusion for separation of transudative from exudative fluid. Sensitivity of light criteria is very high to determine exudative pleural effusion (98%). However, the ability of these criteria for ruling out of transudative effusions is low. For this reason, this study was carried out to determine the level of NT-proBNP in pleural fluid.Methods: A descriptive-analytic study was carried out on 21 patients with complaints of shortness of breath and diagnosis of pleural effusion. Pleural fluid was tapped in these patients and the following tests were performed: LDH, total protein, albumin, cell count, cell differentiation, cytology for malignant cells, ADA, smear for AFB, gram smear and culture.The results of all experiments were analyzed using SPSS V16.Results: Mean age of participants was 65 years. Male and female frequencies were 52.4 and 47.6, respectively. 33.3% of patients had CHF, 28.5% TB, 19.4% malignancy, 4.76% hydatid, and the rest left without diagnosis. A pleural fluid in 66.7% of participants was exudative and in 33.3% was transudative. The levels of NT-proBNP (Pg/ml) in serum and pleural fluid of patients with CHF were 11288.42 and 11036.81, but in malignant patient were 1721.68 and 713.59, respectively, and the levels of NT-proBNP in serum and pleural fluid in TB patient were 2429.30 and 2810.08, respectively. Also, there was no significant difference between the levels of serum and pleural effusion NT-proBNP in transudative and exudative fluid but the level of NT-proBNP was significantly higher in CHF patients compared to others.Conclusions: The results showed that the levels of NT-proBNP in serum and pleural fluid of cardiac patients are higher than other patients, but no significant difference in NT-proBNP between transudative and exudative pleural effusion.

Abstract Background: Pleural mesothelioma (PM) can be difficult to diagnose due to a combination of the growth pattern of the tumour, the invasive nature of the biopsy procedure and the frailty of the patient. Some patients with PM develop a build-up of fluid around their lungs as their first clinical symptom. This fluid can make breathing difficult and is routinely drained as part of standard care. Currently, a cytological test can be performed to identify if PM cells are present in the fluid, however, while this is a specific test, it lacks sensitivity. We examined if (a) cell-free DNA (cfDNA) originating from tumours affecting the lung could be detected in this fluid and (b) if this cfDNA could be determined as coming from PM compared to non-small cell lung cancer (NSCLC), due to changes in genes that only occur in these types of tumours. Methods: Using the QIAamp MinElute ccfDNA Kit, we were able to isolate sufficient concentrations of cfDNA from pleural effusions (n=80) derived from patient samples procured by our colleagues at Southmead Hospital with either PM or NSCLC to perform gene panel sequencing. Using the QIAseq Targeted DNA Pro Human Lung Cancer Research Panel, with additional booster primers covering BAP1 and SETD2, we performed the library preparation in-house. Data analysis was carried out using the QIAGEN CLC Genomics Workbench, with the specific workflows for the gene panels and results were filtered by QC, proportion of variant reads and variant read count. Data were compared to expected variant frequencies published in TCGA. Results: Tumour-specific cfDNA was isolated from pleural effusions in the majority of samples tested. In a proportion of these, variants indicative of a specific type of tumour (PM or NSCLC) were readily detectable, at slightly lower than the expected frequencies than reported in TCGA. Conclusions and future work: This approach can act as a 'rule-in' clinically diagnostic test for the presence of PM or NSCLC in a higher proportion of people that can currently be identified through cytology testing. These individuals could be spared from receiving an invasive biopsy to diagnose their tumour. We are currently working on (i) increasing the sensitivity of this approach to identify more cases and (ii) if a bioinformatic method can identify tumour-specific copy number alterations from the gene panel sequencing data. Citation Format: Connie MacKinnon, Dr Colette Mustard, Dr Nicole Brace, Dr Beth Sage, Jenny Symonds, Colin Nixon, Dr Anna Bibby, A/Prof Antonia Pritchard. Identification of tumour-specific variants from pleural effusion cell-free DNA as diagnostic markers of cancers affecting the lungs [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A060.

Background- Cervical cancer is the commonest cancer among women in developing countries. Screening methods like cervical cytology has brought down the incidence of cervical cancer. AgNOR represents Nucleolar organizer regions stained with silver and are related to cell proliferation rate and tumor malignant potential. The present study was aimed at evaluating the diagnostic role of AgNOR in cervical carcinogenesis. Material and Methods- The present study was conducted in the Pathology department of Gajra Raja Medical College, Gwalior from January 2019 to May 2020 on 500 patients who attended the Gynae OPD. Women of suspected cervical pathology and who underwent Pap smear test followed by subsequent biopsies were included. Results- AgNOR count in chronic cervicitis was 2.72±0.32, in CIN I was 4.02±0.38, in CIN II was 5.47±0.21 in CIN III was 5.59±0.18, in Well differentiated KSCC was 6.48±0.40 and in adenocarcinoma was 6.93±0.21. The study showed that the mean AgNOR count per nucleus was signicantly higher in CIN and malignant cervical lesions. Conclusion- AgNOR count is reproducible, simple, efcient and inexpensive method which can be used as an adjunct to histopathology and cytology especially in differentiating doubtful cases of CIN.

An exact identification of malignant cells in fluid by cytological examination is a well-known diagnostic challenge. One of the common problems is to distinguish reactive mesothelial cells from malignant cells. Conventional smears reported as ‘suspicious for malignancy’ indicate that the suspicious cases could not be classified with certainty as to whether they were reactive mesothelial cells or malignant cells. It poses problem in clinical staging of tumor, treatment and prognosis of malignancy. The purpose of the study was to determine the role of modified method of AgNOR staining in the evaluation of benign and malignant effusions. This cross-sectional study was conducted in the Department of Pathology, BIRDEM General Hospital, Dhaka, from July 2019 - June 2021. A total of 115 cases of effusion were included. All the samples were centrifuged and then smears were prepared from the deposit followed by staining with Hematoxylin &amp; Eosin stain, Papanicolaou stain and AgNOR stain. At first the diagnosis was made on conventional smear method. Then the findings were compared and analyzed by modified AgNOR staining method. In malignant cells, the mean AgNOR count was 5.59±1.05 (±SD) and the AgNORs were multiple and irregular in shape. On the other hand, in benign cells the AgNORs were comparatively larger, single dots with a mean count of 1.31±0.48.The AgNOR count method has definite role in differentiating benign from malignant effusion. This method has supportive value which can be utilized in differentiating malignant effusions from the benign ones, especially in suspicious cases.

AbstractBackgroundMalignancy in pleural effusion is an indication of poor prognosis. The distinction between malignant cells and reactive mesothelial cells in effusion cytology is sometimes difficult and requires ancillary techniques. Evaluation of morphological indicators of chromosomal instability (CI) like micronuclei (MN), chromatin bridging (CB), nuclear budding (NB), and multipolar mitosis (MM) on routine cytology smears is a promising tool to distinguish malignant from benign ascitic fluids. However, it has been scarcely evaluated in pleural effusions. The present study was conducted to evaluate the diagnostic value of these markers in differentiating between malignant and benign pleural fluids.MethodsIt is a cross sectional study in which a total of 72 pleural fluid samples over a period of 2 years received in the cytology department of the hospital were evaluated. The cytological analysis was done by two independent cytopathologists and interpreted as either malignant or benign. Four morphological markers of CI were counted in the May‐Grünwald Giemsa (MGG) stained smears of all the cases and the score was compared with the conventional cyto‐morphological diagnosis.ResultsOut of 72 cases, there were 42 malignant and 30 benign effusions on cytological examination. The mean score of micronuclei count, nuclear budding, chromatin bridging and multipolar mitosis in malignant effusions were 7.26 ± 2.74, 9.55 ± 5.53, 1.83 ± 1.17, and 2.21 ± 1.62 respectively that was significantly higher than the benign effusions (1 ± 0.71, 1.1 ± 0.86, 0.38 ± 0.50, and 0.15 ± 0.37 respectively) (p &lt; .05). On Receiver operating characteristic (ROC) curve analysis, a cut‐off of 5 for the MN count had a sensitivity of 88% and specificity of 100% in detecting malignant pleural effusion [Area under curve (AUC) 95.8%, p &lt; .001].ConclusionEvaluation of morphological indicators of CI on routine MGG stained smears is a simple and cost‐effective method to differentiate between benign and malignant pleural fluids.

AbstractBackgroundRapid and accurate diagnosis of septic peritonitis is critical for initiating appropriate medical and surgical management.ObjectivesThe aim of this study was to determine the diagnostic utility of the total nucleated cell count (TNCC), absolute neutrophil count, neutrophil percentage, and total protein (TP) to distinguish septic versus non‐septic peritoneal effusions in dogs.MethodsElectronic medical records were retrospectively searched for peritoneal fluid samples from 2008 to 2018 and classified as septic or non‐septic based on bacterial culture and/or cytology results. Receiver operator characteristic curves (ROCs) were used to describe the overall diagnostic utility of each test, with optimal cutpoints analyzed to dichotomize continuous variables. Positive and negative likelihood ratios were calculated at these cutpoints.ResultsA total of 166 unique samples, including 87 septic and 79 non‐septic peritoneal effusions, were included. There were no significant differences in dog sex, age, or days hospitalized between groups. Septic effusions had significantly higher TP, TNCC, absolute neutrophil count, and neutrophil percentage compared with non‐septic effusions. The area under the curve of the ROC curves was TNCC (0.80), absolute neutrophil count (0.80), neutrophil percentage (0.64), and TP (0.63). For TNCC and absolute neutrophil count, optimal cutoffs were 17.13 × 103 cells/μL and 19.88 × 103 cells/μL, resulting in positive and negative likelihood ratios of 2.39 and 0.28 and 2.85 and 0.28, respectively.ConclusionsTotal nucleated cell counts and absolute neutrophil counts aid in the differentiation of septic and non‐septic peritoneal effusions with similar diagnostic utility but are not sufficiently sensitive or specific to use without concurrent microscopic evaluation.

AimsPrimary effusion lymphoma (PEL) is a rare aggressive, human herpesvirus‐8 (HHV8)‐associated neoplasm of post‐germinal centre B cell origin. It usually presents as a serous effusion in human immunodeficiency virus (HIV)‐positive patients. PEL is rarely reported in HIV‐negative patients.Methods and resultswWe report seven cases of HIV‐negative elderly men diagnosed with PEL in a single institution. Clinical information and laboratory characteristics were collected. All patients were men, with a mean age of 76 years (range = 60–93) and presented with pleural effusions (n = 6), pericardial effusion (n = 1) and/or ascites (n = 2); two patients had multiple effusions. Extracavitary tissue involvement was present in one patient, who was also a liver transplant recipient. All patients had a decreased blood lymphocyte fraction, with a zero CD4+ count in one. The tumour cells in cytology of effusions showed a moderate amount of cytoplasm, perinuclear hof (a focal area of clearing) and irregular nuclear outlines with coarse chromatin and prominent nucleoli. Immunohistochemically, PEL cells were positive for HHV8 latent nuclear antigen (7 of 7), CD45 (3 of 3), CD30 (4 of 4), MUM1/IRF4 (2 of 2) and were negative for CD3 and CD20 in all seven cases. CD138 was positive in six of seven cases. Epstein–Barr virus (EBV) was detected in two of seven cases by in‐situ hybridisation. B cell clonality by polymerase chain reaction (PCR) was positive in two cases with adequate materials available. Conventional cytogenetic analysis showed complex karyotypes in three of five cases, with recurrent +8, +12 and t(4;12)(q27;q21), and one case with +7. Six of seven patients died of disease with a mean survival of 5.4 months (range = 0.4–11.2 months).ConclusionsPEL can arise in immunocompetent, older patients, in this series all men, and behaves aggressively. These neoplasms are similar to their HIV‐positive counterparts with anaplastic cytomorphology, HHV8 infection and a plasmablastic immunophenotype. The aetiology of PEL is uncertain, but may be related to physiological immunodeficiency associated with ageing.

Abstract Introduction Rheumatoid arthritis (RA) is a systemic inflammatory disorder, with various pulmonary manifestations, including parenchymal disease (interstitial lung disease), inflammation of the pleura (thickening and effusions), and inflammation of the airways and pulmonary vasculature (vasculitis and pulmonary hypertension). These changes may reflect chronic immune activation, increased susceptibility to infection, or direct toxicity from disease modifying or biological therapy. This case report describes a 69-year-old male with longstanding seropositive rheumatoid arthritis who developed severe pneumonia during adalimumab treatment, with subsequent persistent right basal pleural effusion. Treatment with abatacept has resulted in remission with significant decrease in size of the pleural effusion. Case description This patient attended the difficult asthma clinic and also ENT. Nasal polyps, troublesome post-nasal drip and persistent sinus infections led to 2 polypectomies and surgical turbinate resection. Both rheumatoid factor and anti-CCP positive, he was treated with myocrisin, distamine and methotrexate sequentially, without satisfactory response. He was commenced on adalimumab 40mg subcutaneously fortnightly and achieved remission for 22 months. Over a 6-week period he experienced increasing dyspnoea and productive cough. Adalimumab was omitted. Despite three oral antibiotics he continued to deteriorate and developed sepsis (white cell count 13, CRP 277) with an acute kidney injury. Chest radiograph showed right middle lobe infiltrates. CT chest showed no evidence of pulmonary fibrosis or bronchiectasis. Sputum was negative including for atypical infection. Interferon Gamma Release Assay (IGRA) was positive; sputum and urine culture for tuberculosis (TB) were negative after 6 weeks. After the acute episode, repeat contrast CT chest demonstrated residual inflammatory changes in the right lower lobe with a small pleural effusion. Widespread synovitis developed; sulfasalazine was commenced with low dose oral corticosteroids. Interval imaging showed significant increase in effusion. Pleural aspiration confirmed an exudate with a total protein of 77 g/L and an LDH of 2364u/L. Glucose was undetectable and pH low consistent with a cellular metabolically active effusion. No organisms were identified. Thorascopic biopsy revealed fibrinous pleuritis with chronic inflammation, but no evidence of malignancy, ILD, granulomata. Mycobacterial cultures were negative. Over the successive months the patient was managed with sulfasalazine, oral prednisone 5-7.5mg, intra-articular injections and he had a period of relative stability. Joint disease then flared with dramatic synovitis, and a DAS-28 score of 6.46 was documented. Abatacept 125mg subcutaneously weekly was initiated with excellent clinical response. DAS-28 score 2.5 at 6 months, with no infective episodes, and the pleural effusion has significantly reduced in size. Discussion Symptomatic pleural involvement affects 3-5% patients with RA, is more common in older males and those with rheumatoid nodules. Most effusions are unilateral. In this case, effusion persisted after resolution of infection, and the clinical picture was not in keeping with parapneumonic effusion/empyema. Differential aetiologies included rheumatoid, tuberculosis (particularly in the setting of previous anti-TNF) and malignancy. Typically, a rheumatoid effusion is sterile exudate with low pH, low glucose and elevated LDH. Rheumatoid factor may be present. White cell count and cell predominance is variable. Characteristic multinucleated macrophages and necrotic background debris may be seen. Here, the pleural aspiration was not definitive in determining aetiology. Cytology confirmed large numbers of acute inflammatory cells accompanied by macrophages and mesothelial cells. Occasional multinucleated giant cells were seen; these have been described in both tuberculous and rheumatoid effusions. No alcohol/acid fast bacilli were cultured. A sample was sent for adenosine deaminase (ADA). Low ADA excludes tuberculous effusion however it is recognised that in rheumatoid disease it may be elevated. The ADA was elevated at 69 IU/L (0 - 45). If the diagnosis remains unclear, as in this case, histological examination of the pleura may reveal replacement of the normal mesothelial lining with multinucleated giant cells and foci of palisading fibroblasts and lymphocytes surrounding necrotic centres, similar to rheumatoid nodules. This gentleman’s biopsy demonstrated fibrinous pleuritis with chronic inflammation and some patchy mesothelial hyperplasia and no evidence of granulomatous inflammation. The pathologist stated that these appearances are observed in patients with rheumatoid arthritis but are not specific. Most cases of rheumatoid pleuritis improve with treatment of the underlying arthritis; small, asymptomatic effusions don’t require specific intervention. However, persistent effusion can lead to pleural thickening and trapped lung; this would suggest that those with large or persistent effusion should be treated to avoid complication. Key learning points This patient developed a pleural effusion concurrent with sepsis, which persisted post infection and also experienced recurrent chest sepsis (although not as severe at time of admission) over the following 6-9 months. The effusion progressed with time in parallel with joint activity. The persistence of effusion was likely on the basis of RA, and establishing that this was the driving factor once infection/ other aetiology was excluded led to re-evaluation regarding biologic therapy with multi-specialty input. After such significant infection and prolonged period of recovery and investigation, without resolution of the effusion, the patient, the rheumatology and the respiratory teams were each apprehensive about escalating immunosuppressive therapy. Initially, no further treatment with biologic was agreed. However, after several years of moderate ongoing disease activity and then significant flare, the cautious use of biologic agent was reconsidered. At this time, the patient’s quality of life was substantially diminished by his joint disease, and he made an educated decision to consent to treatment with abatacept. The BSR biologic DMARD safety guidelines in inflammatory arthritis recommend using etanercept or abatacept as a first line biologic therapy in patients at high risk of infection, whilst American College of Rheumatology guideline regarding patients with RA and previous serious infection strongly recommends abatacept over TNF-α antagonist. An increased risk of TB has been observed in patients receiving TNF-α antagonist. The British Society for Rheumatology (BSR) guideline regarding biologic DMARD in inflammatory arthritis recommends that all patients should be screened for latent TB with IGRA and chest radiograph, and treated prior to commencing treatment. Conflict of interest The authors declare no conflicts of interest.