Smear Examination Research Articles

Relationship of Knowledge About Cervical Cancer and PAP SMEAR with Early Detection Behavior of PAP SMEAR

<div><table cellspacing="0" cellpadding="0" align="left"><tbody><tr><td align="left" valign="top"><p><em>Cervical cancer is the third leading cause of death in Indonesia. Cervical cancer is usually only diagnosed when the cancer has entered an advanced stage. So the need for early detection as an effort to prevent cervical cancer. One of the most effective and commonly used methods to detect cervical cancer is Pap smear. The purpose of this study was to see the relationship between knowledge about cervical cancer and Pap smear with Pap smear early detection behavior. This study used analytic method with quantitative approach. The population of this study were all married women in Pancoran District, South Jakarta. Then, consecutive non-random sampling technique was used and 153 respondents were obtained with a questionnaire as a tool to obtain research data. Data collection was carried out from September to November 2023. Followed by analysis with the Chi-Square test using SPSS. Based on the results of the study, 58.2% of respondents have good knowledge and 72.5% of respondents have not done Pap smear examination from 158 respondents, there is no significant relationship between knowledge about cervical cancer and Pap smear with Pap smear early detection behavior (p-value=0.093). The conclusion of this study is that there is no relationship between knowledge about cervical cancer and Pap smear with early detection behavior of Pap smear in Pancoran District, South Jakarta. </em><em> </em><em>For further research, it is recommended to provide insight into cervical cancer in depth so that readers are aware of the importance of early detection of cervical cancer</em><em>.</em><strong><em> </em></strong></p><p><strong><em> </em></strong></p></td></tr></tbody></table></div><strong><em>Keyword: Cervival cancer, Knowledge, Pap smear</em></strong>

Роль многокомпонентного комплекса нутриентов для мужчин в рамках преконцепционной подготовки супружеской пары к зачатию.

Introduction. The COVID-19 pandemic has also greatly impacted people's psychological well-being, increasing stress levels, as well as their lifestyle and well-being. Therefore, to maintain health, men should take multicomponent complexes that have a beneficial effect on various organs and systems (including reproductive), have an antioxidant effect and help normalize basic physiological processes in the body. In particular, such supplements can be used as a preconception preparation of a man for conception in a couple. Objective. To evaluate the action of the antioxidant complex «AndroDoz Light» in a group of men in the framework of preconceptional preparation (for conception of a child by a sexual partner). Materials and methods. The data of 30 men aged 23 to 48 years (the average age was 26.3 ± 5.4 years) preparing for conception with a sexual partner were analyzed in this paper. AndroDoz Light was used by men for 1 month: effervescent tablet dissolved in a glass of water (200 ml) once a day during or after meals. To everyone in addition to the spermogram, microbiological examination of the ejaculate, finger rectal examination of the prostate gland (pancreas), ultrasound examination (ultrasound) of the scrotum organs, microscopic examination of the pancreatic secretion (SPH), microscopic examination of the urethral smear and PCR diagnostics of sexually transmitted diseases (androflor) were performed. The concentration of testosterone in the blood, the level of zinc in the sperm plasma were determined. The assessment of the quality of life and sexual function of men was carried out on the IIEF-15 scale (International Index of Erectile Function) and on the NIH-CPSI scale. Results. The multicomponent complex AndroDoz Light helps not only to increase male fertility, improving the functional state of the male reproductive system (qualitative and quantitative spermogram indicators) and morphological indicators: reduces the percentage of sperm with DNA fragmentation, but also improves copulatory function, and is an additional source of necessary nutrients for the male body. Conclusion. The use of the multicomponent complex AndroDoz Light in men as part of the preconceptional preparation of a married couple for conception is accompanied by an improvement in both general well-being and the quality of men's sexual life in general, positively affects the qualitative and quantitative indicators of the spermogram and sperm DNA fragmentation. The results obtained allow us to recommend the use of the multicomponent complex AndroDoz Light in men in the absence of pronounced reproductive problems as part of the preconceptional preparation of a married couple for conception.

Prevalence of Tropical Theileriosis in Cattle through Blood Smear Examination in Sumel District, Duhok Province

Prevalence of Tropical Theileriosis in Cattle through Blood Smear Examination in Sumel District, Duhok Province

Artificial intelligence applications in allergic rhinitis diagnosis: Focus on ensemble learning.

The diagnosis of allergic rhinitis (AR) primarily relies on symptoms and laboratory examinations. Due to limitations in outpatient settings, certain tests such as nasal provocation tests and nasal secretion smear examinations are not routinely conducted. Although there are clear diagnostic criteria, an accurate diagnosis still requires the expertise of an experienced doctor, considering the patient's medical history and conducting examinations. However, differences in physician knowledge and limitations of examination methods can result in variations in diagnosis. Artificial intelligence is a significant outcome of the rapid advancement in computer technology today. This study aims to present an intelligent diagnosis and detection method based on ensemble learning for AR. We conducted a study on AR cases and 7 other diseases exhibiting similar symptoms, including rhinosinusitis, chronic rhinitis, upper respiratory tract infection, etc. Clinical data, encompassing medical history, clinical symptoms, allergen detection, and imaging, was collected. To develop an effective classifier, multiple models were employed to train on the same batch of data. By utilizing ensemble learning algorithms, we obtained the final ensemble classifier known as adaptive random forest-out of bag-easy ensemble (ARF-OOBEE). In order to perform comparative experiments, we selected 5 commonly used machine learning classification algorithms: Naive Bayes, support vector machine, logistic regression, multilayer perceptron, deep forest (GC Forest), and extreme gradient boosting (XGBoost).To evaluate the prediction performance of AR samples, various parameters such as precision, sensitivity, specificity, G-mean, F1-score, and area under the curve (AUC) of the receiver operating characteristic curve were jointly employed as evaluation indicators. We compared 7 classification models, including probability models, tree models, linear models, ensemble models, and neural network models. The ensemble classification algorithms, namely ARF-OOBEE and GC Forest, outperformed the other algorithms in terms of the comprehensive classification evaluation index. The accuracy of G-mean and AUC parameters improved by nearly 2% when compared to the other algorithms. Moreover, these ensemble classifiers exhibited excellent performance in handling large-scale data and unbalanced samples. The ARF-OOBEE ensemble learning model demonstrates strong generalization performance and comprehensive classification abilities, making it suitable for effective application in auxiliary AR diagnosis.

Comparison of Platelet Counts Estimated by Peripheral Blood Smear Examination and Automated Haematology Analyzer

Introduction: Platelet count estimation is one of the common as well as important investigations in clinical practice to diagnose many diseases like dengue, malaria, etc. Different methods for platelet estimations are available. These methods are manual counting semi-automated and automated hematology analyzer counting, platelet count estimation by peripheral blood smear (PBS) method, etc. Semi-automated or automated analyzers may produce erroneous results in the presence of particles or light scatter like giant platelets, fragmented red blood cells, and platelet clumps, so alternative methods like PBS examination can be used for validation. Aim: To compare platelet count estimation performed by the automated cell counter method and the PBS examination method. Objective: Peripheral Blood smear examination acts as a good quality control tool to validate the results produced by the automated cell counter. Materials and Methods: The present study was carried out in the Department of Pathology, Akshaya Health Centre, Urban Bangalore. The study included 100 random blood samples collected into ethylenediaminetetraacetic acid (EDTA) vacutainers over 3 months. These were analyzed by both peripheral blood smear and automated cell counter for platelet estimation. The statistical analysis was done for test performances and their comparisons by using the coefficient of variation (CV), linear regression, and mean differences with SPSS software. Results: No significant difference (p = 0.06635) was observed between the manual peripheral blood smear (PBS) method (platelets average per 100x, multiplied by 15000/ µl) of platelet estimation (207.13 ±15.898 x 1000/ µl) and that of automated cell counter platelet value (206.53 ±16.278 x 1000/ µl). A significant positive correlation was observed between the results of both methods (r=0.9995, p < 0.001) when analyzed by the Pearson correlation test. Conclusion: The peripheral blood smear platelet estimation results are comparable with automated analyzer results. Hence, the PBS examination serves as a quality control tool in assessing the results of the automated cell counters.

Clinical blood test interpretation. Part 1. Neutrophil granulocytes

INRODUCTION. Recent decades are characterized by a rapid introduction of modern automated analytical systems in clinical laboratory diagnostics such as hematology analyzers, which provide a high accuracy of results as well as additional (compared to routine tests) and often more valuable information. Automated blood test has made urgent the problem of interpreting both new tests and “routine” parameters that is relevant to any section of clinical medicine. Therefore, this lecture is aimed at doctors of different specialties.
 OBJECTIVE. Interpret a clinical blood test.
 DISCUSSION. It is hard to review all aspects of hematological analysis within the scope of one lecture, thus part 1 will investigate one of leukocyte populations – neutrophil granulocytes, which include stab and segmented neutrophils. The lecture presents well-known data about neutrophilic leukocytosis and neutropenia. The emphasis is placed on new parameters of automated blood test – IG (immature granulocytes), Neut-RI (reactivity index), Neut-GI (granularity index), MNV (the average volume of neutrophil), structurally dispersed characteristics (NE-WX, NE-SFL, NE-WZ) and some others. Along with modern parameters it was noted the importance of the traditional cytological examination of blood smear, enabling to detect anomalies of neutrophil (degenerative-dystrophic changes of cytoplasm, apoptosis, nucleus pathology).
 Overall, recent years have shown a significant improvement in laboratory diagnostic capabilities of reactive changes (with inflammation, injury, etc.) and blood diseases (anemia, myelodysplastic syndromes, hemoblastosis, etc.), based on accumulated knowledge in the field of laboratory hematology, including clinical significance in evaluating neutrophils.

Macroscopic, cytological and histological diagnosis of proventricular adenocarcinoma in a sulphur-crested cockatoo (Cacatua galerita)

An adult sulphur-crested cockatoo (Cacatua galerita) was unexpectedly found dead and was presented for post-mortem examination. Macroscopically, the proventricular wall was expanded by an infiltrative off-white mass, 10 mm in diameter, with secondary mucosal ulceration. Cytological examination of impression smears revealed a neoplastic epithelial cell population forming occasional well-defined acini with secretory product. Histologically, the neoplastic cells were predominantly arranged in acini and tubules, with fewer in small clusters, supported by a desmoplastic stroma. Neoplastic cells were polygonal, with mostly well-demarcated cell boundaries and a moderate amount of eosinophilic cytoplasm that was multifocally markedly distended by more intensely eosinophilic material. There was an increased nuclear-to-cytoplasmic ratio with moderately sized to large nuclei (approximately 15 μm diameter) with lightly to densely stippled chromatin and single or paired prominent basophilic nucleoli. The mitotic count was 19 in 10 high-power fields (2.37 mm2), with scattered bizarre mitotic figures and atypia including moderate anisocytosis and anisokaryosis and moderate cellular and nuclear pleomorphism. There was frequent intratubular brightly eosinophilic proteinaceous material, which was Alcian blue and periodic acid‒Schiff positive. Proventricular lymphovascular invasion was detected, with metastases within the ventriculus and lungs. The macroscopic, cytological and histological appearance of the neoplasm was consistent with a proventricular adenocarcinoma.

COMPATIBILITY BETWEEN PLATELET HISTOGRAM WITH IP MESSAGE AND PLATELET MORPHOLOGY IN THROMBOCYTOPENIA PATIENTS

Thrombocytopenia is a condition characterized by low levels of platelets in the blood. The platelet count can be checked using a Hematology analyzer, which is an automatic tool for checking a Complete Blood Count (CBC). The principle of this tool is to detect a signal that is comparable to the cell size that has been set on the tool. The results of the CBC examination are accompanied by a histogram and IP Message if there are abnormal cells, so a confirmatory test using a peripheral blood smear is necessary. The purpose of this study was to determine the suitability of the results of the platelet histogram with the IP Message on the platelet morphology of peripheral blood smears of thrombocytopenia patients. This research is a quantitative descriptive study of 54 samples taken using the Saturation Sampling technique at Grati Pasuruan Hospital using the Sysmex XN-1000 from April 1–30, 2023. The variables in this study are histogram results accompanied by IP Messages and platelet morphology on peripheral blood smears. The results of this study are that the histograms that often appear are abnormal heights on the PU Flag and the IP Messages that often appear are “PLT Clumps?”, with a suitable level of platelet morphology on peripheral blood smear examination reaching 90.7%, so it can be concluded that there is a compatibility between the results of the platelet histogram accompanied by the IP Message on the platelet morphology of the peripheral blood smear of thrombocytopenia patients.

Hematologic Abnormalities and Diseases Associated with Moderate-to-Marked Basophilia in a Large Cohort of Dogs.

Basophilia is a rare hematologic finding in dogs. This research aimed to describe the hematologic and clinical characteristics of dogs with moderate-to-marked basophilia. CBC reports with blood smear examinations from dogs presented to the North Carolina State University Veterinary Teaching Hospital were retrospectively reviewed for basophilia (>193 cells/µL). We classified basophilia as moderate when counts were ≥500 cells/µL and marked when they reached ≥1000 cells/µL. We compared the hematologic and clinical profiles of dogs with moderate-to-marked basophilia (the basophilia group) to those without basophilia, serving as our control group. In addition, we investigated differences between dogs with marked basophilia versus those with moderate basophilia, as well as between dogs in the basophilia group with and without concurrent eosinophilia. Diseases associated with moderate-to-marked basophilia included eosinophilic lung disease (p < 0.0001), leukemia/myeloproliferative neoplasms (p = 0.004), parasitic infection (p = 0.004), mast cell tumor (p = 0.005), and inflammatory bowel disease (p = 0.02). Overall, dogs with marked basophilia had a lower frequency of inflammatory diseases (51% vs. 70%, p = 0.009) and a higher frequency of neoplastic diseases (48% vs. 26%, p = 0.003) compared to those with moderate basophilia. In the basophilia group, concurrent eosinophilia was only seen in 36% of dogs. Dogs with concurrent eosinophilia were more often diagnosed with inflammatory diseases (77% vs. 58%, p = 0.006), with fewer diagnoses of neoplasia (19% vs. 40%, p = 0.001), compared to dogs without concurrent eosinophilia. The findings of this study offer veterinary clinicians valuable guidance in determining diagnostic priorities for dogs with moderate-to-marked basophilia.

Studi filariasis berbasis pemeriksaan laboratorium di Kota Semarang

Background: Filariasis is a disease caused by parasitic nematodes of Filariae. Filariasis is caused by filarial worm species, namely Wuchereria bancrofti, Brugia malayi, and Brugia timori. This disease is transmitted through the bite of infected mosquitoes (Anopheles, Aedes, and Culex). Symptoms of filariasis in men or women include damage to the lymphatic system swelling of the arms, legs, and genitals, which causes pain, decreased productivity, and social problems. Filarial diagnosis through examination of blood smears to detect filarial parasites in the blood, accompanied by hematological and clinical chemical examinationsObjective: This study aimed to describe filariasis through blood tests (blood smears, hematological examinations, and clinical chemistry examinations) on filariasis respondents using a cross-sectional approach.Method: This type of descriptive research uses a cross-sectional approach. The research subjects were 6 filarial respondents in Pedurungan District, Semarang City. The examination was conducted at the Tologosari Wetan and Kulon Health Centers on September 12-18, 2020. Secondary data collection was obtained from capillary blood examination using thin blood smears, automatic hematological examination using a hematology analyzer, and clinical chemistry examination using the spectrophotometer method with a spectrophotometer. The research data was processed and analyzed using Univariate analysis using maximum, minimum, and average values.Results: Hematological examination for hemoglobin level of 12,5 g%, leukocytes 3200 cells/ml, platelets 137.00 cells, erythrocytes 4,07x106 cells, lymphocytes 35 cells, neutrophils 73 cells, eosinophils 1 cell, monocytes 9 cells, MCH 30,78 pg, MCV 86,48 fl, MCHC 36,24 g/l and clinical chemistry examination results for cholesterol 145,7 g/dl and triglycerides 104,6 g/dl. Meanwhile, on examination of thin blood smears, microfilariae were found.Conclusion: The results of the blood smear evaluation showed an abnormality in the size of the red blood cells in the form of microcytic (smaller red blood cell size), and filarial findings were found. In contrast, hematology and clinical chemistry examination results obtained normal results.

Prevalence of Tuberculosis Infection and Disease Among Homeless Persons Staying in NGO Shelters in Chennai

Latent tuberculosis infection (LTBI) is a inactive stage after being infected with Mycobacterium tuberculosis and it reactivates into pulmonary tuberculosis (PTB). This study was to estimate the prevalence of LTBI in homeless persons staying in 13 shelters run by NGOs in Chennai metro city, India. A cross-sectional survey was conducted from January to April 2018. All inmates aged 15 years and above were screened for PTB infection using tuberculin skin test (TST) and disease smear and culture examinations. Out of a total of 1,068 registered, 1,016 persons were screened for PTB disease and 818 persons for LTBI. Among them, PTB was detected in two persons and TST was positive in 275 persons. Univariate and multivariate analyses were performed to find out the factors associated with LTBI. It was found that LTBI is significantly associated with male gender, aged more than 40 years and with chest X-ray TB lesions. This study finding highlights that low prevalence of TB disease and one third of homeless persons infected with tuberculosis. Our finding suggests that there is a need for prevention interventions to eliminate TB in this key population.

Cryogenic enrichment of Plasmodium falciparum gametocytes from spiked whole blood

Each individual cell type typically requires a unique set of conditions for optimal cryopreservation outcome, which relates to its specific response to cryoprotective agent (CPA) toxicity, osmotic behavior and sensitivity to ice crystallization. Cryopreservation of heterogenous cell populations is therefore exceedingly difficult as it requires separate and often conflicting conditions for each cell type. Conversely, these contrasting conditions could be utilized to favor cryogenic preference of a single cell population within a heterogenous sample, leading to its enrichment by elimination of remaining cells. To establish proof-of-concept for this overall approach, a protocol was developed for the cryogenic enrichment of Plasmodium falciparum gametocytes from whole blood. To accomplish this goal, we evaluated the effects of CPAs and cooling conditions during cryopreservation of whole blood samples spiked with P. falciparum gametocytes. We identified that cooling to −80 °C at a rate of −1 °C/min in the presence of 11 % glycerol selectively favors recovery of gametocytes. This protocol eliminates 95.3 ± 1.7 % of total blood cells and recovers 43.2 ± 6.5 % of parasites, leading to a 19-fold enrichment as assessed by microscopic examination of blood smears. This protocol is tunable, where gametocyte enrichment 900-fold may be feasible, however there is an apparent tradeoff in overall parasite recovery. Although translation of this protocol for point-of-care testing for malaria presents many challenges, the overall approach of cryogenic purification may prove useful for alternative diagnostic applications.

Disseminated histoplasmosis diagnosed on peripheral blood smear: a case report

Abstract Introduction/Objective Histoplasmosis is an infection caused by the inhalation of a dimorphic fungus, Histoplasma capsulatum. H. Capsulatum var capsulatum is the main variety we encounter in the United States and is endemic in the Mississippi river valley. In immunocompetent individuals the infection is usually asymptomatic or presents as flu- like symptoms. Disseminated histoplasmosis is a severe and often fatal opportunistic infection if not treated, usually seen in immunocompromised patients. Methods/Case Report We report a case of a 27-year-old female intra-venous drug user with past medical history of untreated HIV diagnosed in 2018, who is being admitted for suspected necrotizing fasciitis of the vulva. On admission, her complete blood count shows pancytopenia with a white blood cell count of 2.9 × 109/L (NR 4.3-10.8 × 109/L), hemoglobin 6.1 g/dL (NR 12-16 g/dL), and a platelet count of 43 × 109/L (NR 150-400 × 109/L). A peripheral blood smear obtained showing rare neutrophils with intracellular organism highly suggestive of histoplasma capsulatum. CT Chest showing diffuse ground glass opacities and a subsequent cytology from bronchoalveolar lavage performed revealed numerous macrophages filled with ovoid organism surrounded by a cytoplasmic halo, GMS stain highlights the rim consistent with histoplasma capsulatum. Patient expired after one week in the ICU for severe acute respiratory distress syndrome, sepsis, histoplasmosis, strep pneumonia and hepatic failure. Results (if a Case Study enter NA) NA Conclusion Careful examination of Peripheral blood smear along with correct clinical history can aid in early diagnosis of disseminated histoplasmosis.

Chronic Myeloid Leukemia Presenting with Mixed Phenotype B/Myeloid Blast Phase: A Diagnostic Challenge

Abstract Introduction/Objective Chronic myeloid leukemia (CML) is typically identified in chronic phase, however about 5% of cases present in a more advanced phase. In blast phase (BP), the blasts are most commonly of myeloid lineage, less commonly lymphoid, and rarely mixed lineage cases are encountered. A mixed phenotype at presentation raises the possibility of a de novo mixed phenotype acute leukemia (MPAL) with BCR::ABL1 fusion and requires careful correlation with ancillary studies to confirm the diagnosis. Methods/Case Report The patient is a 60-year-old man who presented with leukocytosis, anemia, normal platelet count, and blasts identified on peripheral blood smear examination. A bone marrow biopsy demonstrated hypercellular marrow and a dimorphic population of large and small blasts, comprising nearly 70% of cellularity. Flow cytometry demonstrated distinct myeloid and B-lymphoid blast populations, both of which expressed CD13 and TdT. Interphase fluorescence in situ hybridization (FISH) showed the BCR::ABL1 fusion in more than 90% of nuclei, including segmented nuclei, and conventional cytogenetic studies showed t(9;22)(q34.1;q11.2) in 19 of 20 metaphases. Molecular studies demonstrated the minor BCR::ABL1 p190 transcript, as well as ASXL1 and dual RUNX1 mutations. The discrepant number of BCR::ABL1 positive cells by FISH and conventional cytogenetic studies compared to the proportion of blasts, and its presence in segmented nuclei, enabled a diagnosis of CML with mixed phenotype blast phase despite absence of features such as basophilia or significant granulocytic left shift. Results (if a Case Study enter NA) NA Conclusion We share this case of CML in mixed phenotype blast phase to highlight the importance of careful evaluation in distinguishing this rare presentation from de novo MPAL, particularly with BCR::ABL1 fusion. Features such as a disproportionate number of nuclei or metaphases carrying the t(9;22) translocation on FISH or conventional cytogenetic studies and the presence of BCR::ABL1 in mature granulocytes can be helpful when other morphologic features of CML are absent.

A Patient with Target Cells and Hyperkalemia - or why Hematologists Should Still be Able to ‘Fly by Sight‘

INTRODUCTION: Hematology has changed. Both benign and malignant hematology rely heavily on new diagnostic techniques, such as flow-cytometry, cytogenetics and most recently single gene analysis by next generation sequencing. Examination of a blood smear has come out of favor and is usually performed by lab technicians and not by the hematologist. This case report describes how examination of the blood smear which showed an unexpected red blood cell morphology eventually led after 40 years to the right diagnosis. CASE: R. C. is a 42-year-old male (100 kg = 220 lbs, 1,83 m = 6 ft, BMI 29,9) diagnosed in childhood with spherocytosis. His medical history was remarkable for that after birth he needed blood exchange for kernicterus (bilirubin-induced brain dysfunction), and since age 3 months he had epileptic seizures. His blood counts were not that abnormal and since he had no or only mild anemia throughout his whole life he received no further w/u for his blood disorder. Another remarkable finding was that he always had hyperkalemia but since he had no symptoms (no palpitations, dyspnea, chest pain, nausea, or vomiting) and nobody found an explanation no further workup was pursued. In November 2022 he presented to a nearby hospital for epileptic seizures. His spleen was mildly enlarged (14,5 x 6,7 cm) and because of his h/o spherocytosis the neurologist sent him for hematologic re-evaluation. His physical was unremarkable except for that the margin of the spleen was palpable. He was neither pale nor icteric. He drinks no alcohol and does not smoke. Medications were lamotrigen, cholecalciferol, and cenobamate. His blood counts are shown in the table. Since his MCHC was normal, which does not fit spherocytosis, we ordered a blood smear shown in the figure. There were no spherocytes but many target cells. Then we sent for an eosin-5′-maleimide (EMA) binding test which was also unremarkable (124% binding, with spherocytosis usually &amp;lt; 80%). However, acidified glycerol lysis test showed increased osmotic fragility (lysis time 45 sec., normal &amp;gt; 1800 sec). This unusual combination together with his hyperkalemia led to the hypothesis of a disorder related to the xerocytosis or stomatocytosis complex. However, the morphologic picture did not fit stomatocytosis. We therefore sent for NGS analysis which revealed a heterozygous mutation of SLC2A1, variant c.857G&amp;gt;A; p.Gly286Asp (stomatin-deficient cryohydrocytosis). Stomatin-deficient cryohydrocytosis is a very rare congenital deficiency in GLUT1 and stomatin with moderate hemolytic anemia, pseudohyperkalemia and various neurologic defects. The occurrence of target cells (but no stomatocytes) with cryohydrocytosis has - to our knowledge - been reported only once before. CONCLUSIONS: In Germany benign hematology no longer plays the role it had 30-40 years ago. This may be due to the fact that congenital anemias and erythrocyte abnormalities occur less frequently in the relatively homogeneous German population, at least less often than in southeastern Europe or in other countries. Also, patients with mild stable anemia that do not require treatment, are often not seen by a hematologist. To the authors' opinion the most relevant change is that blood smear examination, which is reimbursed with 50 cents has fallen out of fashion in daily practice. However, in this case only the blood smear revealed, that the prior diagnosis of spherocytosis could not be correct. There were no spherocytes but target cells. The patient, though, had not the typical factors associated with target cells (no liver disease, no hyperlipidemia, no thalassemia or iron deficiency). Eventually NGS analysis revealed stomatin-deficient cryohydrocytosis. We present this case because it was the blood smear examination that eventually led to the right diagnosis. All hematologists should have a microscope “next door” and even if blood smear examination is reimbursed poorly it is a diagnostic tool still at the core of our profession

Evaluation of Novel Minimal Screening Antibody Panel for Immunophenotypic Detection of Hematological Malignancies in a Resource-Constrained Setting: A Prospective Study

INTRODUCTION Immunophenotyping of abnormal cells by flow cytometer enabled this technology in diagnosing, prognostication and guiding therapy for blood malignancies. International consensus exist for diagnosis of leukemia, lymphoma and plasma cell neoplasm (PCN) 1-3 using flow cytometer, but it is seldom used for screening these diseases. In clinical setting, including our tertiary care center in India, we come across patients whose peripheral blood (PB) smear examination does not hint towards the type of blood-related disorder. We identified the need to screen these patients by flow cytometry before channelling them towards established diagnostic work-up for malignancies. The aim of this study is to evaluate a flow cytometry screening assay, designed in-house, to detect blood malignancies in a resource-limited (sample, time, cost, infrastructure) clinical setting. MATERIALS AND METHODS This prospective double-blinded study was conducted between November 2022 and June 2023 with institutional ethical approval. Remainder of samples sent for routine clinical tests that met our inclusion/exclusion criteria were selected (n=221). Symptomatic patients with blood-related disorder, whose PB did not show any abnormal cells were included. All patients who had blast, lymphocytosis with atypical lymphocytes, lymphadenopathy, B-symptoms, splenomegaly or found to have plasmacytoma/amyloidosis/CRAB features were excluded. Three tube 8 colour 19 antibody panel (Table1) was designed. Immaturity markers and lineage specific markers are combined in a single tube to detect leukemias. CD5, CD10, CD19, CD20, kappa and lambda were put together to identify B-CLPD. A lyse-wash-stain protocol was followed and the samples were analysed in BD FACSCantoII flowcytometer with autofluroscence and compensation correction done for each sample. The findings of our screening assay were validated with morphological and clinical correlation and GraphPad Prism version 8.0.2 was used for statistical analysis. RESULTS Our antibody panel targeted only cell surface antigens, which reduced the cost of the test and permitted the analysis of the sample within 1 hour. During the study period, out of the 396 discard samples obtained, 221 patients met our inclusion criteria (Figure1). We analysed these samples with our screening assay and detected 51 patients with blood malignancies that included 19 acute leukemias, 11 lymphomas and 21 PCN and 167 samples with no evidence of leukemia, lymphoma or PCN which were subsequently diagnosed with anemias, platelet disorders, drug- or disease-induced leukocytosis/leukopenias and organ dysfunction not involving blood or bone marrow. The number of samples studied and the computed statistical parameters are given in Table2. Representative bivariate dot plots are given in Figure2. Out of the 221 test results, 214 had clinical, morphological and screening test concordance. One case of PCN could not be identified by morphological examination, where screening assay could identify clonal plasma cell population. Another one case was clinically diagnosed as PCN, but missed on screening panel because we did not see a definite cluster of CD38 + cells showing light chain kappa/lambda restriction. The remaining 5 samples were dilute in nature. In 3 out of 5 dilute samples, screening panel could identify the abnormality which was concordant with clinical diagnosis (based on clinical, biochemical, imaging, molecular and cytogenetics) where morphology failed. The other 2 cases were identified morphologically on bone marrow imprint. This flow cytometry screening assay was found to have overall 94.4% sensitivity, 100% specificity, 100% positive predictive value and 98.2% negative predictive value, delineating the efficacy of this minimal antibody screening panel in detecting blood malignancies. CONCLUSION Compared to the existing screening panels, this novel screening test has comprehensive set of antibodies (19 only) to detect leukemia, lymphoma and PCN simultaneously within 1 hour and at a lower cost. In patients screened negative, further blood malignancy-related testing may be avoided, thereby reducing the total cost incurred to the patient. The usefulness of this screening test is limited with dilute sample. However, the present study shows that this screening panel can be used in small resource-constrained and remote settings with a trained flow cytometry technologist alone.

Development and Pilot Testing of a Comprehensive Mobile Application to Assist Cell Count Determination During Peripheral Smear and Bone Marrow Examination.

In the modern era of complete blood count analysis, manual differential count is performed whenever 'flags' are generated by an automated hematology analyzer. Traditionally, tally counters with five or eight keys are used for manual differential count. A few mobile applications are available to perform this task; however, the application features and cell representation are limited. The primary objective of our study was to develop an indigenous, comprehensive mobile application to assist with manual blood cell differential count. The secondary objective was to measure the usability of a newly developed application among undergraduate medical students. A new mobile application was developed using a Java development kit, Version 11.0.13 (Oracle Corporation, Austin, USA) in Android Studio Dolphin (2021.3.1) (Google, California, USA). The application content was validated by three pathologists with more than five years of experience. The app's usability was tested among 60 participants using a validated mHealth App Usability Questionnaire (MAUQ). The questionnaire had 18 items covering three domains: ease of use, interface & satisfaction, and usefulness. The newly developed application supports peripheral smear WBC differential count, platelet count, reticulocyte count, malaria parasite quantification, and bone marrow differential count. During usability testing, the app was easy to use in 95% (57/60) of participants, time-efficient in 91.7% (55/60), and helpful for healthcare practice learning in 96.7% (58/60). The total mean score was 6.11, indicating high usability. A comprehensive mobile application to assist manual differential count with adequate cell representation was developed. The mobile application was easy to use, time-efficient, and valuable among the study participants.

Case Series: Mixed Infectious Keratitis by Pythium insidiosum and Fungal Species.

This case series is the first to illustrate mixed infection from Pythium sp. and fungal species in corneal ulcer. This case series aimed to alert all toward the possibility of both Pythium sp. and fungal species infection in case of nonresponding corneal ulcer treated with either antifungals or antipythium drugs alone. Increased suspicion of mixed infection in case of nonresponding fungal/ Pythium keratitis may facilitate early and prompt management. Six patients presented with signs of either fungal or Pythium keratitis. They underwent ophthalmological examinations, smear examinations, cultures, and polymerase chain reaction (PCR). Therapeutic penetrating keratoplasty was performed in cases where symptoms worsened after treatment with either antifungal or antipythium drugs. The half corneal button (HCB) was shared for histopathological and microbiological examinations. In the first case, smear examination from corneal scraping (CS) revealed Pythium -like filaments, which were confirmed with PCR; however, Aspergillus nidulans grew in culture. In the second case, iodine-potassium iodide (IKI) staining was positive for Pythium ; however, PCR was positive for both Pythium and fungus, which was further confirmed by DNA sequencing. In the third case, IKI staining and HCB were positive for Pythium ; however, PCR was positive for fungus, which was identified as Candida saitoana with DNA sequencing. In the fourth case, Pythium grew in the CS culture; however, Candida sp. grew in the HCB culture. In the fifth case, Cladosporium sp. grew in culture from CS; however, Pythium insidiosum grew from the anterior chamber exudate after therapeutic penetrating keratoplasty. In the sixth case, smear examination revealed septate fungal filaments, and Cladosporium sp. grew in culture; however, HCB on histopathological examination showed features of Pythium keratitis. In unresponsive cases of Pythium or fungal keratitis, diagnostic modalities such as IKI and PCR should be implemented as a routine practice, in addition to smears and cultures.

A RESPON KEMOTERAPI FLUDARABINE PADA HAIRY CELL LEUKEMIA

HCL is part of a rare B-cell lymphoproliferative disorder and is a subtype of Chronic Lymphocytic Leukemia. It is characterized by progressive pancytopenia, splenomegaly, and bone marrow hypercellularity. Patients often come because of abnormal complete blood count results or complications such as infection. This case report explains a case of a 41-year-old woman who had recurrent shortness of breath and cough as symptoms, where the patient had a history of acute myeloblastic leukemia (AML) with non-routine fludarabine chemotherapy. After receiving treatment and underwent further investigations in the form of peripheral blood smear and bone marrow aspiration examination, it was found that the patient had hairy cell leukemia. Fludarabine was given to the patient and it is reported to give effect partial remission (based on last bone marrow aspiration) along with pancytopenia as a clinical manifestation from myelosuppression as a side effect for long-term used fludarabine.&#x0D; HCL is part of a rare B-cell lymphoproliferative disorder and is a subtype of Chronic Lymphocytic Leukemia. It is characterized by progressive pancytopenia, splenomegaly, and bone marrow hypercellularity. Patients often come because of abnormal complete blood count results or complications such as infection. This case report explains a case of a 41-year-old woman who had recurrent shortness of breath and cough as symptoms, where the patient had a history of acute myeloblastic leukemia (AML) with non-routine fludarabine chemotherapy. After receiving treatment and underwent further investigations in the form of peripheral blood smear and bone marrow aspiration examination, it was found that the patient had hairy cell leukemia. Fludarabine was given to the patient and it is reported to give effect partial remission (based on last bone marrow aspiration) along with pancytopenia as a clinical manifestation from myelosuppression as a side effect for long-term used fludarabine.

Quality of blood smear examinations for malaria parasites in Chenzhou City after malaria elimination

To analyze the quality of blood smear examinations for malaria parasites in Chenzhou City, so as to provide insights into sustainable consolidation of malaria elimination achievements. All positive blood smears from fever patients were irregularly sampled from each county (district) of Chenzhou City from 2018 to 2022 and reexamined, and no less than 3% negative blood smears were reexamined. The preparation, dyeing, cleanliness and microscopic examination results of blood smear were reexamined, and the quality of blood smear reexaminations was assessed using a descriptive statistical method. A total of 13 625 fever patients received blood smear examinations for malaria parasites in Chenzhou City from 2018 to 2022, of which 21 were positive and 13 604 were negative; 687 blood samples were reviewed, and the percentage of negative blood smear reexaminations was 4.90% (666/13 604), with a 63.51% rate of qualified negative blood smears preparation, a 67.87% rate of qualified dyeing and a 76.13% rate of qualified cleanliness, and no missing diagnosis found. There were 21 positive blood smears reexamined, and the proportions of qualified blood smears preparation, dyeing and cleanliness were all 85.71%, with 2 smears mistaking Plasmodium species (9.52%). The percentage of qualified negative blood smears preparation was 51.41% in 2022, which reduced by 31.61% in relative to that (75.17%) in 2019 (χ2 = 9.033, P < 0.05), and the percentage of qualified negative blood smears dyeing was 60.19% in 2022, which reduced by 28.82% in relative to that (84.56%) in 2019 (χ2 = 19.498, P < 0.05), while the percentage of qualified negative blood smears cleanliness was 62.96% in 2022, which reduced by 28.93% in relative to that (88.59%) in 2019 (χ2 = 23.826, P < 0.001). In addition, there were no significant differences in the proportion of qualified negative blood smears preparation (χ2 = 0.260, P > 0.05) or dyeing (χ2 = 1.094, P > 0.05) among the three years, while a significant difference was detected in the percentage of qualified negative blood smears cleanliness (χ2 = 12.175, P < 0.05). No missing diagnosis was seen in blood smear examinations for malaria parasites among fever patients in Chenzhou City after malaria elimination; however, there were reductions in proportions of qualified blood smears preparation, dyeing and cleanliness. Quality control of blood smear examinations is recommended to be reinforced in key regions of Chenzhou City.