May-Grunwald Giemsa Research Articles

UTILITY OF AUTOMATED HEMATOLOGY ANALYZER IN THE DETECTION OF MALIGNANT CELLS IN SEROUS EFFUSIONS

Abstract Introduction/Objective Although microscopic examination is the gold standard for body fluid malignant cytology, it has a higher turnaround time and inter observed variability. Screening of serous effusions for malignant cells using haematology methods of automated analysis provides a rapid and objective result to the clinicians for initiating necessary treatment. Automated haematology analysers are equipped with body fluid (BF) mode that can detect high fluorescence cells body fluid (HF-BF) cells, mononuclear and polymorphonuclear (PMN) cells using flowcytometry laser scatter principles. The high fluorescence parameters useful for identifying malignant cells in the effusions Methods/Case Report The study was performed between February 2023to Februart 2024 at AIG Hospitals, Hyderabad after obtaining approval from the medical ethics committee of thehospital. Body fluidswere collected in K2 EDTA vaccutainer. Simultaneous preparation of smears for microscopic examination and morphometry wasdone and the smears were stained using Romanowsky stain (Leishmann & Giemsa). Cytospin smears were prepared by cyto- centrifugation (Sakura cyto- tix). The slides were stained by Hematoxylin and eosin, May Grunwald Giemsa (MGG) and Papanicolaou (PAP) stains for morphological differentiation and detection of malignant cells. Cell blocks were prepared after through mixing of the fluid using direct sedimentation method and the slides were stained by haematoxylin and eosin stain. IHC was performed where ever necessary. Results (if a Case Study enter NA) A total of 258 fluid samples comprising of 223 ascitic and 35 pleural fluids. Medical statistical software version 20.2 was used for statistical analysisof the results. Considering fluid cell block analysis as the gold standard the patient population was divided into 2 groups, malignant (8 samples) and non- malignant cell group(250 samples). Cytological examination of cytospins smears has shown to have the highest specificity, PPV and NPV in the detection of malignant effusions. Conclusion Our study shows that cytology and histopathological examination of body fluids remains the gold standard for the detection of malignant cells. The HF-BF parameter acts as a rapid, useful tool to predict the presence of malignant cells in serous effusions and select samples for microscopic reviewing that would have otherwise gone undiagnosed. We believe that with defining cut off values for the HF-BF parameter may be used as an adjunct diagnostic tool.

Robustness assessment of an automated AI-based white blood cell morphometric analysis system using different smear preparation methods.

Numerous AI-based systems are being developed to evaluate peripheral blood (PB) smears, but the feasibility of these systems on different smear preparation methods has not been fully understood. In this study, we assessed the impact of different smear preparation methods on the robustness of the deep learning system (DLS). We collected 193 PB samples from patients, preparing a pair of smears for each sample using two systems: (1) SP50 smears, prepared by the DLS recommended fully automated slide preparation with double fan drying and staining (May-Grunwald Giemsa, M-G) system using SP50 (Sysmex) and (2) SP1000i smears, prepared by automated smear preparation with single fan drying by SP1000i (Sysmex) and manually stained with M-G. Digital images of PB cells were captured using DI-60 (Sysmex), and the DLS performed cell classification. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were used to evaluate the performance of the DLS. The specificity and NPV for all cell types were 97.4%-100% in both smear sets. The average sensitivity and PPV were 88.9% and 90.1% on SP50 smears, and 87.0% and 83.2% on SP1000i smears, respectively. The lower performance on SP1000i smears was attributed to the intra-lineage misclassification of neutrophil precursors and inter-lineage misclassification of lymphocytes. The DLS demonstrated consistent performance in specificity and NPV for smears prepared by a system different from the recommended method. Our results suggest that applying an automated smear preparation system optimized for the DLS system may be important.

Clinicopathological Spectrum of Head and Neck Lesions By FNAC at a Tertiary Care Centre in Barpeta, Assam – A Retrospective Study

Background: Various neoplastic and non-neoplastic lesions arising from lymph nodes, salivary glands, thyroid gland, soft tissue and metastatic deposits comprises the wide spectrum of lesions in the head & neck region. The Foundation of Head and Neck Cancer (FHNC) helps in differentiating non-neoplastic lesions from neoplastic and further from benign and malignant lesions. The aim of this study was to evaluate the role and diagnostic accuracy of FNAC in diagnosis of palpable head and neck lesions. Materials and methods: A total of 360 cases were included in the study. Out of the 360 cases, 122 (33.89%) cases were sent for histopathological examination and the results were correlated histologically. Aspirations were done by using 10 ml syringe and 22/23-gauge needles. Smears were stained with May Grunwald Giemsa (MGG). Cytomorphological diagnosis was given. Results: Out of 360 patients of head and neck lesions studied, lymph node (40.56%) was the predominant site aspirated with reactive lymphoid hyperplasia being the commonest lesion. Soft tissue constitutes 31.11% cases followed by thyroid lesions with 19.72% cases, salivary gland 7.22% cases, and oral cavity 1.39% cases. FNAC was inconclusive in 4.17% cases. Conclusion: Head and neck lesions are common lesions located superficially that can be easily diagnosed on cytology. Although excisional biopsy is the gold standard for diagnosing head and neck malignant lesions, FNAC plays a vital role as a quick and inexpensive diagnostic procedure with high diagnostic accuracy. It distinguishes between inflammatory/infection and neoplastic lesions, avoiding needless procedures and expediting therapy. FNAC is a low-cost, accurate diagnostic method that may be used as a first-line examination.

Diagnosis of Septic Body Cavity Effusion in Dogs and Cats: Cytology vs. Bacterial Culture

The elective test for the determination of the effusions etiopathogenesis is represented by physico-chemical analysis and cytology. Nevertheless, the bacterial culture and antibiotic sensitivity tests are crucial for setting therapy and for the outcome. This study compared cytology with microbiology in the etiologic diagnosis of exudative body cavity effusions in dogs and cats collected from October 2018 to October 2022. All samples underwent aerobic and anaerobic culture and cytology examination. Bacterial identifications were confirmed using matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry, whereas cytological samples were blindly evaluated either in May Grunwald-Giemsa (MGG) or Gram-stained samples by two board-certified clinical pathologists. A moderate agreement (κ = 0.454) between cytology and bacterial culture was revealed. The sensitivity of the cytological evaluation in our study ranged from 38.5% to 67.9%, and the specificity ranged from 88.9% to 100%, depending on the type of the effusion, so cytology may not be representative of the etiopathogenesis, whereas bacterial culture can misidentify or fail to isolate the correct pathogen for difficult in vitro growing due to the presence of inhibitory substances or contamination. Cytology and bacterial culture results for exudative body cavity effusions in dogs and cats can be misleading if conducted separately, so these two tests should be performed together to increase diagnostic accuracy.

Adult niemann-pick disease: A case report

Adult niemann-pick disease: A case report

Pathological Responses in Asian House Shrews (Suncus murinus) to the Naturally Acquired Orientia tsutsugamushi Infection.

Scrub typhus is a re-emerging disease caused by Orientia tsutsugamushi, transmitted by mites belonging to the family Trombiculidae. Humans and rodents acquire the infection by the bite of larval mites/chiggers. Suncus murinus, the Asian house shrew, has been reported to harbor the vector mites and has been naturally infected with O. tsutsugamushi. The present study aimed to localize and record O. tsutsugamushi in the tissues and the host response in shrews naturally infected with O. tsutsugamushi. Sheehan's modified May-Grunwald Giemsa staining was carried out in 365 tissues from 87 animals, and rickettsiae were documented in 87 tissues from 20 animals. Immunohistochemical (IHC) staining, using polyclonal antibodies raised against selected epitopes of the 56-kDa antigen, was carried out, and 81/87 tissue sections were tested positive for O. tsutsugamushi. By IHC, in addition to the endothelium, the pathogen was also demonstrated by IHC in cardiomyocytes, the bronchiolar epithelium, stroma of the lungs, hepatocytes, the bile duct epithelium, the epithelium and goblet cells of intestine, the tubular epithelium of the kidney, and splenic macrophages. Furthermore, the pathogen was confirmed by real-time PCR using blood (n = 20) and tissues (n = 81) of the IHC-positive animals. None of the blood samples and only 22 out of 81 IHC-positive tissues were tested positive by PCR. By nucleotide sequencing of the 56-kDa gene, Gilliam and Karp strains were found circulating among these animals. Although these bacterial strains are highly virulent and cause a wide range of pathological alterations, hence exploring their adaptive mechanisms of survival in shrews will be of significance. Given that the pathogen localizes in various organs following a transient bacteremia, we recommend the inclusion of tissues from the heart, lung, intestine, and kidney of reservoir animals, in addition to blood samples, for future molecular surveillance of scrub typhus.

Blood count abnormalities in the association of sickle cell disease and malaria in clinical hematology at the CNHU-HKM in Cotonou (Bénin)

Although a protective effect of hemoglobin S has been described, malaria has frequently been associated with increased morbidity and mortality in sickle cell disease patients in Africa. Various cytopenias are frequently found on the haemograms of these patients. In Benin, a malaria-endemic zone with a high prevalence of sickle cell disease, the aim of this study was to establish and compare the blood count profile according to hemoglobin type in the association of sickle cell disease and malaria. This was a prospective descriptive study. It covered a 24-month period from October 2020 to October 2022. It included all patients with major sickle cell syndrome seen in clinical haematology and with a positive thick drop/parasite density, whatever the parasitaemia value. For each patient, a blood count was performed on the Sysmex XT 4000i machine, supplemented by a smear study after staining with May-Grunwald Giemsa. Data were analyzed using R 3.6.1 software. Three hundred non-redundant cases with a positive thick smear were identified in sickle cell patients, including 208 SS homozygotes (69.3%) and 92 SC heterozygotes (30.7%). In contrast, there were 181 non-redundant cases with a negative thick smear, including 119 SS homozygotes (65.7%) and 62 SC heterozygotes (34.3%). Among subjects with a positive thick smear, the majority of patients (70%) exhibited clinical symptoms. Severe malaria was observed in 58% of the cases. The proportion of severe malaria was higher in SS homozygote patients than in double heterozygote SC patients (p < 0.0001). The mean parasite density was higher in SS individuals (4 320.7 ± 2 185 trophozoites/pL) compared to SC individuals (1 564.4 ± 1 221 trophozoites/pL; p < 0.0001). Plasmodium falciparum was the only species identified. The mean hemoglobin level in impaludated SS subjects was 6.1 g/dL, significantly lower than that in non-impaludated SS subjects (p < 0.0001). The average white blood cell count in impaludated SS subjects was 16.58 G/L, compared to 13.2 G/L in those with a negative thick smear (p < 0.0001). Twenty cases of thrombocytopenia were found in SS subjects with a positive thick smear, compared to 6 cases in those with a negative thick smear. As for SC subjects with a positive thick smear, the average hemoglobin levels and white blood cell counts were 9.8 g/dL and 10.63 G/L, respectively, compared to 11.27 g/dL and 7.3 G/L in SC subjects with a negative thick smear. Eighteen cases of thrombocytopenia were found in subjects with a positive thick smear, compared to 17 cases in those with a negative thick smear. Sickle cell disease and malaria represent two major public health problems. However, contrary to popular belief, sickle cell disease is not immune to malaria infestation. Malaria is recognized as one of the main causes of morbidity and mortality in sickle cell patients, particularly children. In Benin, its association with sickle cell emergencies has already been reported.Our study found that malaria was predominantly associated with the homozygous SS form (p < 0.00001). Severe malaria was the most common clinical form. All malaria infestations in our series were due to Plasmodium falciparum, and parasitaemia was significantly higher in SS patients (p < 0.0001).The hematological profile of the association of sickle cell disease and malaria in homozygous SS individuals in our series showed characteristics of a normocytic normochromic anemia with neutrophil-predominant leukocytosis. Compared to non-malaria-infected SS individuals, there was a significant worsening of anemia, neutrophil-predominant leukocytosis, and a decrease in the average platelet count. In SC individuals, there was rather a microcytic normochromic regenerative anemia associated with neutrophil-predominant leukocytosis. Compared to non-malaria-infected SC individuals, there was a significant decrease in the rate of anemia and neutrophil-predominant leukocytosis. Anemia is a constant feature in homozygous sickle cell disease, and the low values recorded illustrate the hemolytic nature of malaria, especially in SS individuals, and the better tolerance of SC individuals. Furthermore, the low baseline hemoglobin levels make SS individuals more vulnerable to malaria-induced anemia compared to SC individuals. The observed leukocytosis is generally accompanied by reticulocytosis in the case of major sickle cell syndrome, which must be taken into account for result validation. It is the expression of compensatory bone marrow reaction to anemia and inflammatory mechanisms resulting from malaria infestation. Finally, thrombocytopenia was significantly more common in SC patients, even though they were adults living in malaria-endemic areas. Malaria can frequently induce thrombocytopenia through platelet consumption during the "rosetting" phenomenon. In SS patients, the effects of "rosetting" could be compensated for by the bone marrow stimulation induced by anemia. In our series with adult subjects living in an endemic area, thrombocytopenia is not a frequent biological disturbance. In a clinicalbiological context combining a systemic inflammatory response syndrome with anemia and neutrophil-predominant leukocytosis in a SS or SC sickle cell patient, the clinician should be able to consider malaria and confirm or rule out this diagnosis.

Evaluation of bronchoalveolar lavage fluid from donkeys using four different cytological stains: A pilot study

Bronchoalveolar lavage fluid (BALF) cytology is used for the diagnosis of non-infectious lower airway inflammation in equids. Discrepancies have been reported in the differential cell count when different staining methods were used both in humans and horses. The objective of this study was to compare the results of BALF cytology in donkeys using four different staining methods: modified May-Grunwald Giemsa (mMGG), Diff-Quick (DQ), Toluidine blue (TB) and Perls Prussian blue (PPB). Nine healthy Amiata female donkeys were enrolled. The BAL procedure was performed as previously described and pairs of cytocentrifuged BALF slides were stained with each method. No differences between mMGG and DQ were found for macrophages, neutrophils, and eosinophils, while differences were found in mast cell count between DQ vs.TB, but not between mMGG vs. DQ or mMGG vs. TB. Finally, no differences were obtained in the differential count for hemosiderophages comparing mMGG, DQ and PPB. The mMGG appears to be an excellent stain for the identification of all possible cell types, including mast cells in the BALF of donkeys. DQ, if used alone, may lead to inappropriate identification of mast cells. These results are consistent with the literature on BALF staining methods in horses.

Utility of guided FNAC and cell block preparation from liver and gall bladder masses: Learning experience from a tertiary care center.

Ultrasound- and CT-guided fine needle aspiration cytology (FNAC) increases the accessibility of intra-abdominal masses to the liver and gall bladder with the advantages of low cost and high diagnostic yield. Cell block technique has been known for further increasing the diagnostic accuracy. We aimed to study the effectiveness of FNAC and the cell block method in cytological diagnosis of liver and gall bladder masses. We also followed a step-wise approach to increase the success rate. A 2-year observational study was done from July 2020 to June 2022. Total 80 guided (CT and ultrasound) aspirations were done from space occupying/mass lesions in the liver [74 (92.5%)] and gall bladder [6 (7.5%)], out of which cell blocks by the plasma thrombin method were prepared in 12 cases (15%). The on-site radiological details were noted, and rapid on-site evaluation was done in 65 cases (81.25%). The prepared cytology slides were stained with Papanicolaou, H and E and May-Grunwald Giemsa (MGG) stain. The cytological diagnosis was noted, and the uses and limitations (if any) were observed in each case. A step-wise structured questionnaire format was developed to assist the reporting pathologist so as not to miss out on important diagnostic observations, if present. FNAC in 71 cases (88.7%) gave a conclusive diagnosis. The maximum number of cases were of adenocarcinoma [38 (51.3%)] from the liver followed by hepatocellular carcinoma in 10 cases (13.5%). In gall bladder masses, all 6 cases (100%) were positive for malignancy, out of which 4 cases (66.7%) could be characterized as adenocarcinoma. The cell block preparation was helpful in reaching the diagnosis as well as typing the malignancy in 10 cases (83.3%). The chief limitation observed on conventional cytology smears was inadequate cellularity, which caused inconclusive diagnosis in 9 cases (11.25%). The reporting questionnaire was helpful chiefly in terms of time-efficient reporting in 34 cases (42.5%), increasing the ease and confidence in 69 cases (86.25%) and the advantage of reproducibility of data in all cases (100%) according to the case-by-case evaluation by the reporting pathologists. Guided FNAC in conjunction with the cell block technique is extremely helpful in the evaluation of mass lesions of the liver and gall bladder for cytological diagnosis. A proper step-wise approach may be useful to reach a quick and effective diagnosis.

Expanding the VEXAS diagnostic workup: the role of peripheral blood cytological analysis.

VEXAS syndrome is a newly described autoinflammatory entity characterized by somatic mutations in the UBA1 X-linked gene in hematopoietic progenitor cells. Several studies have demonstrated that the presence of vacuoles in progenitor cells from bone marrow aspirates is a hallmark finding for this syndrome. Therefore, this study aimed to characterize leukocytes from VEXAS patients versus patients with ANCA-associated vasculitis (AAV), familial Mediterranean fever (FMF), and healthy donors (HD) to define a specific cytological pattern that can support VEXAS diagnosis. Twelve VEXAS patients were included in the study. Blood samples from FMF (n = 16), AAV (n = 16) and HDs (n = 20) acted as controls. May-Grünwald Giemsa (MGG) staining was used for studying cellular morphology, including cytoplasm, granules, and vacuoles and to perform a cytogenic evaluation of leucocytes. Plasma IL-1β, IL-1α, TNFα, IL-18 and IL-8 were measured using ELISA assay. The cytological analysis from blood smears confirmed the presence of immature neutrophils in VEXAS patients. We found a greater number of vacuoles in VEXAS patients vs. FMF, AAV and HD. Micronuclei (MNi) and cell death rate were higher in VEXAS patients vs. HD. Cell death correlated with IL-1β and IL-8 levels. MNi were positively associated with IL-8 and IL-1β levels, and with the percentage of immature neutrophils and vacuoles. In conclusion, our findings suggested that cytological test may be supportive for VEXAS diagnosis, despite genetical analysis is mandatory for confirming the disease. Finally, we identified several cytological hallmarks that may distinguish the VEXAS "cytotype" not only from HD but also from other inflammatory diseases.

Staining, magnification, and algorithmic conditions for highly accurate cell detection and cell classification by deep learning.

Research into cytodiagnosis has seen an active exploration of cell detection and classification using deep learning models. We aimed to clarify the challenges of magnification, staining methods, and false positives in creating general purpose deep learning-based cytology models. Using 11 types of human cancer cell lines, we prepared Papanicolaou- and May-Grünwald-Giemsa (MGG)-stained specimens. We created deep learning models with different cell types, staining, and magnifications from each cell image using the You Only Look Once, version 8 (YOLOv8) algorithm. Detection and classification rates were calculated to compare the models. The classification rates of all the created models were over 95.9%. The highest detection rates of the Papanicolaou and MGG models were 92.3% and 91.3%, respectively. The highest detection rates of the object detection and instance segmentation models, which were 11 cell types with Papanicolaou staining, were 94.6% and 91.7%, respectively. We believe that the artificial intelligence technology of YOLOv8 has sufficient performance for applications in screening and cell classification in clinical settings. Conducting research to demonstrate the efficacy of YOLOv8 artificial intelligence technology on clinical specimens is crucial for overcoming the unique challenges associated with cytology.

Morphological markers of chromosomal instability as predictors of malignancy in pleural effusion.

Malignancy 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. It 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. Out 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 < .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 < .001]. Evaluation 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.

Antiviral effect of high-pressure nasal stimulation.

In a previous study, we reported an increase of nasal nerve growth factor (NGF) in patients treated with high-pressure administration of sterile saline isotonic solution (HPpSIS). Herein we characterized the nasal mucosa in terms of innate immune response and cytokine signature, including antiviral properties. Potential NGF and antiviral benefits of HPpSIS were also discussed. Twenty (20) patients (11 males, 9 females; age range 30-75 years old) underwent HPpSIS and nasal samples were collected before and after treatment. Nasal scraping was used for morphological (smears and Quick May-Grunwald Giemsa staining, MGG), biochemical (Histamine, Serotonin; ELISA) and molecular (messenger RNA, mRNA) analyses. Amplification of transcripts specific for Toll-like receptor (TLR) 3 (TLR3), TLR7, TLR9, Interleukin-(IL) 18 (IL18), IL13, IL12, eosinophil-derived neurotoxin (EDN), Eosinophil Cationic Protein (ECP), γ Interferon (γIFN), tryptase and serotonin was performed using the 2-step real-time Reverse Transcription Polymerase Chain Reaction (RT-PCR). Clinical and laboratory data were analyzed and compared. The clinical evaluation showed a protective effect of our therapy. Smears showed the presence of leucocytes, eosinophils (EOs) and mast cells (MCs), and increased immunoreactivity for ECP/RNase3 and EDN after HPpSIS. ELISA showed increased levels of Serotonin and EDN associated with unchanged levels of substance P(SP) and histamine. Increased eosinophil-derived neurotoxin eosinophil-derived neurotoxin (EDN) levels were confirmed by in situ fluorescent analysis. HPpSIS induced the upregulation of TLR3, TLR7 and TLR9 transcripts, while no changes were observed for Intercellular Adhesion Molecule 1 (ICAM1), IL18, Interleukin-15 (IL15) and IL12 transcripts nor for Interleukin-6 (IL6) and IL13. No changes were also observed for γIFN and EDN/RNase2 transcripts, while ECP/RNase3 transcripts were significantly upregulated after HPpSIS. Finally, tryptase transcripts were unchanged while serotonin transcripts were significantly increased after HPpSIS. The clinical and biomolecular changes observed at the nasal mucosa due to HpSS treatment suggest the activation of an innate surveillance, by means of TLR transcription, and a possible anti-viral response due to EDN upregulation. It remains to be verified if NGF, known to be released locally upon HpSIS treatment, might in part be responsible for this local activation.

Usefulness of imprint cytology for early diagnosis in vertebral lesions

Abstract Background: Vertebral pathologies can be divided into degenerative, infective, or neoplastic. Infective and neoplastic lesions can progress rapidly and may lead to compression of the cord. Thus, rapid diagnosis for these lesions is important. Frozen section and fine-needle aspiration cytology techniques are less useful in hard bony tumors. Imprint cytology has been successfully used in the diagnosis of soft tissue tumors. The purpose of this study was to evaluate the role of imprint cytology in the early diagnosis of various spinal pathologies. Materials and Methods: Thirty-eight patients with suspected diagnosis of infection or tumor underwent transpedicular biopsy using Jamshidi needle. At least two imprint slides were made using core tissue obtained from biopsy. These slides were, thereafter, stained with the May-Grunwald–Giemsa method. The core biopsy sample was also sent for histopathological examination, after fixing it with 10% formalin. Paraffin-embedded blocks and slides were made for histopathology reporting, as per institution’s protocol. All imprint cytology slides were screened and reported by same pathologist. Results: The sensitivity of imprint cytology was 68.6%. Specificity, positive predictive value, negative predictive value, and overall accuracy of the imprint cytology were 100%, 100%, 21.4%, and 71%, respectively, when compared to histopathology. The mean duration of imprint cytology reporting was only 36 min while that for histopathological report was 15 days. Conclusion: Imprint cytology is a simple and rapid method for obtaining diagnosis in suspected vertebral lesions of infection or tumor. It is a reliable and cost-effective method in experienced hands. Although imprint cytology shows modest correlation in the ability to identify positive results, it forms a good diagnostic tool in confirming true negative cases with high diagnostic efficacy.

Cytomorphologic visualization of circulating tumor cells in urinary bladder cancer patients using ScreenCell™ technology: Potential as a simple cytology test.

Circulating tumor cells (CTC) are a recent technique which is a potentially important prognostic factor in many solid tumors. There are many techniques of detecting CTCs, but they usually implement costly techniques like EpCAM targeted detection, fluorescence-based diagnosis, or magnetic bead based positive or negative selection. The diagnostic utility of simple cytomorphological diagnosis after routine staining of CTCs have been rarely studied. We aimed to detect CTCs in 24 patients clinically suspected to have Urinary Bladder Cancer using a simple but efficient patented filtration technology (ScreenCell™), followed by optical microscopic visualization after routine May-Grunwald-Giemsa (MGG) staining. The detected CTCs were then tested for association with the histologic type, lamina propria invasion, deep muscle invasion and the T-stage. Out of the 24 patients tested, one was found to have papilloma, nine had low grade urothelial carcinoma, 13 had high grade urothelial carcinoma and one had poorly differentiated adenocarcinoma. Of these, two LGUC, eight HGUC and one adenocarcinoma had detectable CTC. Presence of CTCs had a statistically significant association with Lamina propria invasion (P = .006) and T-stage (P = .02), and a trend toward significance for differentiating LGUC from HGUC (P = .10). These results suggest that cytomorphological detection of CTC is likely to be clinically useful in diagnosis and prognostication of urinary blader cancers. These findings need to be confirmed on studies with larger sample sizes.

Patterns of salivary gland lesions on fine needle aspiration cytology (FNAC)

A diverse range of inflammatory, cystic and neoplastic processes affect salivary glands.Fine needle aspiration cytology (FNAC) is a reliable tool for evaluation of salivary gland lesions. To analyse the cytomorphological features of salivary gland lesions and to assess the frequency of distribution of these lesions. It was an observational study conducted in the Department of Pathology, Govt. Medical College, Jammu. FNAC was performed on patients presenting with salivary gland swelling. May-Grunwald-Giemsa (MGG) and Papanicolaou (PAP) stained smears were examined under light microscope. The cytomorphological features were analysed and diagnosis was made. There were 65 cases which included 31 (47.7%) males and 34 (52.3%) females. The mean age of patients was 42.96 years. On microscopic examination, neoplastic lesions were 35 (53.8%) and non neoplastic lesions were 30 (46.2%). Pleomorphic adenoma was the most common benign neoplastic lesion and also was the most frequently diagnosed lesion in our study (21, 32.3%). Mucoepidermoid carcinoma cases were maximum among malignant lesions (4, 6.2%). Parotid gland was involved in majority of cases. FNAC is a simple, rapid, reliable tool for the evaluation of salivary gland lesions thereby helping clinicians in deciding the further management plan.

Role of fine needle aspiration cytology in the diagnosis of cutaneous and subcutaneous swellings

A wide range of inflammatory as well as neoplastic lesions can present as cutaneous and subcutaneous swellings all over the body. FNAC (Fine Needle Aspiration Cytology) is considered to be the first reliable technique for diagnosing them. To assess the cytological spectrum of lesions presenting as skin and subcutaneous swellings. It was an observational study conducted in the Department of Pathology, Govt. Medical College, Jammu. FNAC was performed on patients presenting with palpable skin and subcutaneous swellings. May-Grunwald-Giemsa (MGG) and Papanicolaou (PAP) stained smears were examined under light microscope. The cytological features were studied and diagnosis was made. The present study had 290 patients which included 169 (58%) males and 121 (42%) females. On FNA examination, 111 (38.3%) were non-neoplastic and 179 (61.7%) were neoplastic. Epidermal inclusion cyst was the most common non- neoplastic lesion. Among neoplastic lesions, 166 cases (57.2%) were benign and 13 cases (4.5%) were malignant. Lipoma was the most common benign neoplastic lesion and was also overall the commonest lesion in our study. Metastatic carcinoma cases were maximum among malignant lesions. Fine Needle Aspiration Cytology is a rapid, reliable technique for initial assessment of cutaneous and subcutaneous swellings and thereby guiding further management.

Role of FNAC in the diagnosis of lymph node malignancies - An institutional experience

Lymphadenopathy is a sign of underlying infection, primary or metastatic malignancy. Fine-needle aspiration is a simple and reliable tool for its initial investigation. To analyse the cytomorphological features of malignant lymph node lesions and to determine the frequency of different lymph node malignancies. It was an observational, retrospective study conducted in the cytology section of Department of Pathology, Govt. Medical College, Jammu. It included lymph node aspirates that were reported as malignant. May-Grunwald-Giemsa (MGG) and Papanicolaou (PAP) stained smears were examined under light microscope and the cytological features were studied. A total of 97 malignant lymph node aspirates were included in this study. Metastatic malignancy accounted for maximum number of cases (78, 80.4%) followed by lymphoma (19, 19.6%). Squamous cell carcinoma was the most common metastatic malignancy and also the most frequently diagnosed malignant lesion in our study (42, 43.3%). Cervical lymph nodes were the most commonly aspirated lymph nodes.FNAC of lymph nodes is a simple and valuable tool in the diagnosis of lymph node malignancies.

Refined Diagnosis of Pleural Effusions by Immunocytochemistry of Cell Blocks.

The main objective of microscopic examination of pleural effusions is to ascertain the presence of malignant cells. Effusions prepared routinely using May-Grünwald-Giemsa (MGG)- and Papanicolaou (PAP)-staining can, in a number of cases, provide inconclusive cytological results regarding malignancy. This report describes the refined diagnosis of such cases based on immunocytochemical analysis of pleural effusion cell blocks. Of the 340 pleural effusions obtained during 2019 at the Department of Clinical Cytology, Gävle Hospital, Sweden, 63 (18.5%) contained atypical cells of undetermined significance or potentially malignant cells. This diagnosis could be refined using Epithelial Cell Adhesion Molecule/EPCAM (BEREP4) immunocytochemical analysis of effusion cell blocks, allowing previously inconclusive effusions to be classified as clearly benign 42/63 (66.7%) or malignant 21/63 (33.3%). Effusions initially diagnosed as clearly malignant (27/340; 7.9%) were all 27 (100%) BEREP4-immuno-stained. Most BEREP4-positive effusions (37/48; 77.1%) were also carcinoembryonic antigen (CEA) positive. The number of BEREP4-positive cells, however, tended to exceed that of CEA-positive cells. The BEREP4 positive effusions were further examined using different monoclonal antibodies, such as Thyroid transcription factor 1 (TTF-1) for primary pulmonary adenocarcinoma, to determine the original site of the primary tumour. Immunohistochemical staining of pleural effusion cell blocks significantly refines the diagnosis of serous pleural effusions, especially in cases where the preliminary diagnosis was atypical cells of undetermined significance or potentially malignant cells. Furthermore, in the cases of malignancy, the origin of the primary tumour could most often be determined.

Evaluation of Intraoperative Imprint Cytology and Frozen Section in Determination of Tumor and Tumor Margins

Intraoperative pathology consultation is often required for guiding immediate surgical decisions so as to limit the extent of surgery. Accurate intraoperative diagnosis is crucial. The present study evaluates the accuracy of imprint cytology and frozen section in intraoperative consultation for tumor and tumor margins. A total of 157 tissue specimens for type of lesion, tumor margins and lymphnode metastasis were included. Imprint was taken by holding the tissue bit with forceps and touching it onto a clean glass slide without undue pressure or lateral movements. They were stained with Papanicolaou, May-Grunwald Giemsa and Hematoxylin and Eosin [H&amp;E] stains. The tissue bits taken were then embedded and frozen. Section cutting was done using cryostat and were stained using rapid Haematoxylin and Eosin stain, and examined under a light microscope. Later, the tissue was fixed in 10% neutral buffered formalin for routine histopathological processing then stained with H&amp;E stain. Based on the histological diagnosis, the accuracy of intraoperative imprint cytology and frozen section were evaluated. A total of 157 tissue specimens were received during the study period, which were classified into specimens for type of tumor [145 specimens], tumor margins [54 specimens] and lymph node metastasis [7 specimens] for analysis. The overall diagnostic accuracy of imprint cytology in detection of type of tumor was 97.9% and that of frozen section was 98.6%. It can be concluded that imprint cytology typically provides the accurate diagnosis without the need for a sophisticated machine such as cryostat which is required for frozen section analysis.