Clinical Pathology Department Research Articles

Inoculation of pleural fluid in blood culture bottles could be a routine practice in pleural infection

Background Worldwide, pleural infections are becoming more common. Overuse of empirical antibiotics has been linked to longer hospital stays and higher death rates, causing the emergence of antibiotic-resistant pathogens. So, proper care requires the identification of the pathogenic bacteria using pleural fluid culture. In addition to normal culture, pleural fluid inoculation in blood culture bottles enhances the proportion of patients with detectable infections in less time. Aim To assess whether inoculating pleural fluid into a blood culture bottle will increase the yield of isolated pathogens compared to routine culture techniques in evaluating pleural infection. Also, to compare the time of pathogen detection when comparing the detection by blood culture bottle versus the routine microbiological culture technique. Recognize the distribution of isolated bacteria from our center and their susceptibility profile to different antibiotics and predict the most effective empirical antibiotics. Patients and methods Cross-sectional analytical prospective study conducted at the Department of Pulmonology and Clinical Pathology in Cairo University Hospitals. It included 70 patients with infected pleural effusion. Under ultrasound supervision, pleural fluid was extracted and transported to the laboratory for routine microbiological culture. At the same time, an automated blood culture bottle was inoculated with 5–10 ml to be incubated. All the detected pathogens were identified and tested for antimicrobial susceptibility according to CLSI. Results The mean age of our patients was 47.1 ± 15.6 years. Empyema was documented in 16 (22.9%) of our patients, whereas 54 (77.1%) had exudative pleural effusion. Positive aerobic infection in blood culture bottle was seen in 37 patients, compared to 21 patients in direct pleural culture. A statistically significant difference was found between the culture positive rate in aerobic blood culture bottles and direct pleural culture (P=0.001). Conclusion Blood culture bottle inoculation of infected pleural fluid increases the sensitivity of microbial yield in a shorter time than standard culture.

Assessment of the tissue factor pathway inhibitor (TFPI-rs7586970 variant) polymorphism as a risk factor in myocardial infarction patients

Context The tissue factor pathway inhibitor (TFPI), a Kunitz-type serine protease inhibitor, inhibits the extrinsic pathway of pathways, and might significantly contribute to the pathogenesis of coronary artery diseases. Aim To assess the TFPI-rs7586970 genetic variant as a risk factor in patients with myocardial infarction (MI). Settings and design A descriptive case–control study in the Premises of the Department of Clinical Pathology and Cardiology in Suez Canal University Hospitals included 156 individuals; 78 MI patients and 78 individuals as a control group. Methods and material An interview questionnaire, clinical and laboratory assessment, and molecular genotyping of the TFPI-rs7586970 genetic variant were done using real-time PCR Eva-Green-based high-resolution melting curve (HRM) analysis. Informed consent was taken from all the participants and the study protocol was approved by the Ethics Committee, Faculty of Medicine, Suez Canal University. Statistical analysis used For statistical analysis, t-test, Mann-Whitney U test, chi-squared-test (χ2), one-way ANOVA (analysis of variance), Kruskal-Wallis test, and binary logistic regression model were used. Results The MI group recorded 26 allele copies of the TFPI-rs7586970 genetic variant whereas the control group recorded six copies. The binary logistic regression model for MI occurrence showed that the TFPI-rs7586970 genetic variant polymorphism is a significant predictor for the development of MI. Conclusions TFPI-rs7586970 genetic variant polymorphism is a significant risk factor for the development of MI.

Pathophysiology, Histopathology, and Differential Diagnostics of Basal Cell Carcinoma and Cutaneous Squamous Cell Carcinoma-An Update from the Pathologist's Point of View.

Basal cell carcinoma (BCC) and cutaneous squamous cell carcinoma (cSCC) are the most frequently occurring non-melanocytic skin cancers. The objective of our study is to present the pathophysiology of BCC and cSCC and its direct relationship with the histopathological diagnostics and the differential diagnostics of these types of cancer, based on the morphological characteristics, immunohistochemical profile, and genetic alterations. The qualitative study was based on emphasizing the morphological characteristics and immunohistochemistry profiles of BCC and cSCC and the differential diagnostics based on the tissue samples from the Clinical Pathology Department of Mures Clinical County Hospital between 2020 and 2022. We analyzed the histopathological appearances and immunohistochemical profiles of BCC and cSCC in comparison with those of Bowen disease, keratoacanthoma, hyperkeratotic squamous papilloma, metatypical carcinoma, pilomatricoma, trichoblastoma, Merkel cell carcinoma, pleomorphic dermal sarcoma (PDS), and melanoma. Our study showed the importance of the correct histopathological diagnosis, which has a direct impact on the appropriate treatment and outcome for each patient. The study highlighted the histopathological and morphological characteristics of NMSCs and the precursor lesions in HE and the immunohistochemical profile for lesions that may make the differential diagnosis difficult to establish.

Comparison of Special Stains for Analysing the Morphology of Sperm: A Cross-sectional Study

Introduction: In India, male infertility constitutes 40% of infertility cases in males, which has become a serious problem in developing countries. Numerous studies and advanced techniques have been identified in assessing sperm morphology. The detection of sperm morphology is a simple and widely accepted method for determining sperm viability. It assists in selecting appropriate treatments for male infertility in assisted reproductive techniques. Infertility issues can arise in healthy males due to altered sperm morphology. Aim: To analyse the morphology of sperm using basic histopathology stains such as Haematoxylin and Eosin (H&E), Giemsa (G), Eosin-Nigrosin (EN), and Papanicolaou (Pap) stains. Materials and Methods: A cross-sectional study was conducted from April 2022 to June 2022 at the Department of Clinical Pathology, Sree Balaji Medical College and Hospital, Chennai, Tamil Nadu, India. Thirty semen samples were collected from healthy males and stained with H&E, Giemsa, Eosin-Nigrosin, and Papanicolaou stains. The results were observed and tabulated based on sperm shape, size, and motility using Kruger’s strict morphology method. A comparison was made to identify the fastest and most cost-effective method among the four stains. Descriptive statistics were used for the comparison. Results: Among the 30 semen samples, H&E and Papanicolaou stain methods were found to be rapid and cost-effective for analysing sperm morphology compared to the other stains (p-value <0.001). The findings included cases of Normozoospermia (18), Oligozoospermia (9), Necrozoospermia (1), and Teratozoospermia (2). In the Papanicolaou stained samples, the mean sperm head length (μm) was 8.92a±0.5, head width (μm) was 4.48a±0.33, head perimeter (μm) was 27.69a±1.85, head area (μm²) was 33.96a±3.74, tail length (μm) was 44.31a±2.02, sperm length (μm) was 53.24a±2.18, with no cytoplasmic droplets greater than half the size of the head and two vacuoles observed. Papanicolaou stain was the least expensive stain (9.50rs) compared to the other stains used for assessing sperm morphology. Conclusion: Papanicolaou stain is a simple and cost-effective method for analysing sperm morphology compared to other special stains. However, EN Stain is used to detect the morphology of live sperm. Sperm morphology assessment is necessary to monitor the causes of male infertility and determine appropriate treatment modalities.

Diagnostic Utility of Bone Marrow Aspiration, Trephine Biopsy, and Flow Cytometry in the Evaluation of Various Haematological and Non Haematological Disorders: A Cross-sectional Study from Northern India

Introduction: The spectrum of haematological and non haematological disorders is vast in various age groups. Complete blood counts and other routine laboratory tests are not always sufficient to diagnose these diseases. Bone marrow examination plays an important role in diagnosing their underlying causes. Aim: To analyse the spectrum of various haematological and non haematological disorders reported in Bone Marrow Aspiration (BMA) and compare them with Bone Marrow Trephine Biopsy (BMB) and Flow Cytometry (FCM) findings where applicable. Materials and Methods: A one-year cross-sectional study was conducted in the Department of Clinical Pathology, PGIMS, Rohtak, Haryana, India from April 2022 to March 2023. A total of 518 consecutive BMA samples were morphologically analysed. Comparative evaluations were performed among BMA, BMB, and FCM where applicable. Diagnostic accuracy was calculated, and the findings of discordant cases were tabulated. Definitive diagnosis of lymphoma/leukaemia and Paroxysmal Nocturnal Haemoglobinuria (PNH) cases were made using FCM. Results: The patients’ ages ranged from three months to 86 years, with a mean age of 38.4 years. The male to female ratio was 1.3:1, with a slight male predominance. The highest number of cases were of anaemia {183 (35.3%) and 164 (31.7%), respectively} and leukaemia {128 (24.7%) and 134 (25.9%), respectively}, followed by normal marrow studies {39 (7.5%) and 32 (6.2%), respectively} and megakaryocytic thrombocytopenia {24 (4.6% in each} in both BMA and biopsy. Among anaemia cases {183 and 164 cases in BMA and BMB}, the majority were of the megaloblastic type {62 (33.8%) and 54 (32.9%), respectively}, followed by hypoplastic/aplastic type {40 (21.8%) and 50 (30.5%), respectively}. In cases of leukaemia (128 and 134 cases in BMA and BMB), acute leukaemia cases (76 (59.4%) and 82 (61.2%), respectively} outnumbered chronic leukaemia cases {52 (40.6%) and 52 (38.8%), respectively} in both BMA and BMB. The concordance and discordance rate between BMA and BMB were 87.6% (419 cases) and 12.4% (59 cases), respectively. Diagnosis was exclusively made by BMB in cases of myelofibrosis, granulomatous disease, and Hodgkin’s lymphoma. Conclusion: BMA cytology is a relatively safe and mildly invasive technique for evaluating various haematological and non haematological disorders with better preservation of cellular morphology. However, in cases with dry/blood taps and focal marrow involvement, BMB should be performed, as it shows well-preserved marrow architecture with all its cellular and stromal components. FCM is a definitive diagnostic modality for further categorisation of acute leukaemia and Chronic Lymphoproliferative Disorders (CLPD).

SOCIALIZATION OF CORRECT HANDWASHING STEPS TO CREATE A HEALTHY STUDENTS AT DENANYAR, JOMBANG, INDONESIA

Elementary school students in Indonesia generally do not know the proper and correct steps for washing hands according to the World Health Organization (WHO), which is indicated by the high rate of spread of hand-borne diseases, including diarrhea. Diarrhea is an endemic disease in Jombang Regency with the incidence rate increasing in 2019 compared to 2018. Teaching Staff of the Department of Clinical Pathology, Faculty of Medicine, UNAIR, carry out community service activities with the aim of providing education on how to wash hands properly and correctly for Madrasah Ibtidaiyah students (MI) Mambaul Maarif Denanyar Jombang. The method of socializing the correct hand washing steps is carried out in two stages, namely education and evaluation. The education carried out is an explanation of the correct steps for washing hands according to WHO, what germs are found on human hands, what diseases can be transmitted through hands, and when we should wash our hands. Evaluation is carried out by culturing germs from the surface of the hands at two times, namely before and after washing hands. The data obtained is the difference in bacterial culture results in specimens before and after washing hands. The growth of bacteria before hand washing showed more colonies compared to after hand washing.

Epidemiological and histological characteristics of cutaneous squamous cell carcinoma and its precursor lesions – A single-center study

Abstract Objective: Cutaneous squamous cell carcinoma (cSCC) is a skin malignancy that is one of the non-melanocytic skin cancers (NMSCs). The objective of our study was to highlight the epidemiological and histological characteristics of cSCC diagnosed in a clinical county hospital. Methods A retrospective cross-sectional study was performed of histopathologically diagnosed cases of cSCC from the clinical Pathology Department of the Mures Clinical County Hospital, Târgu Mureș, Romania. We included 96 cases that were diagnosed between January 1, 2017, and December 31, 2020. Results: Of the 96 cases included in the study, 82 were identified as cSCC, 5 as Bowen Disease, and 9 as keratoacanthoma. The majority of the cases were diagnosed in 2018 (n = 30; 31.25%) and 2019 (n=36; 37.50%). The median age of the patients was 63.0 years. Slightly over half of the patients were male (n=50; 52.08) and 49 patients (51.04%) grew up in urban areas. Forty-six cases (56.10%) were well differentiated; 25 (30.49%) moderately differentiated, and 11 (13.41%) poorly differentiated. Almost all of the lesions (93; 96.88%) were removed within the safety excision margins. Conclusion: Most of the patients were diagnosed with cSCC in 2018 and 2019 and were over 70 years old. The majority were males who grew up in urban areas. Even though most of the lesions were well differentiated and completely excised surgically, the differential diagnoses between cSCC and other skin malignancies were made based on the morphological aspects of the lesions, followed by an immunohistochemical profile when necessary.

Correlation between Serum Levels of Vaspin, BMI, and Waist Circumference on Obese and Non-Obese Subjects

Obesity is the accumulation of excessive fat resulting from an imbalance in energy intake compared to energy used for a long period. Vaspin is secreted in adipose tissue and may exhibit compensatory responses related to obesity, insulin resistance, and type 2 diabetes mellitus. This study aimed to determine vaspin levels in obese and non-obese subjects. An observational research design with a cross-sectional approach was used in this study. A total of 80 research subjects were included in this study. The research was conducted at the Clinical Pathology Laboratory at the Department of Clinical Pathology, Hasanuddin University Hospital in January 2022. Kolmogorov-Smirnov test was used to determine the normality of the data. Chi-Square, unpaired T-test, Mann-Whitney test, and Spearman test were used as statistical tests. This study showed a significant difference in anthropometric parameters between the obese and non-obese groups (p < 0.001). There was a significant difference in the mean levels of vaspin between the obese and non-obese groups (707.68±774.47 ng/mL vs. 510.60±460.08 ng/mL, p=0.004). There was a positive correlation between vaspin levels and waist circumference (r = 0.245, p=0.004) and between vaspin levels and BMI (r = 0.322, p=0.028). The serum levels of vaspin tended to be higher in obese subjects compared to non-obese subjects. The positive correlation between vaspin levels and BMI and waist circumference indicated that a higher BMI and/or waist circumference led to a higher vaspin level.

Comparison of Various Blood Vacuum Tubes Brand for Clinical Chemistry, Hormonal, Hematology, and Hemostasis Parameters

Link of Video Abstract: https://youtu.be/O7-fCHyvGo4 Background: Laboratory tests are integral to the medical decision-making process. Blood vacuum tubes need to be validated to increase the analytics quality. This study aimed to compare various vacuum tube brands that use serum clot activator and serum separator tube (SST) for clinical chemistry and hormonal parameters, K3EDTA anticoagulant for hematological parameters, and citrate for hemostasis parameters. Methods: The comparative study was carried out cross-sectional in the Department of Clinical Pathology, Dr. Cipto Mangunkusumo National Public Hospital, on June 20, 2020. There were 20 subjects for each type of tube. The control tube was BD Vacutainer®, and the test tubes were Samplix®, VACUETTE®, Vacusera®, and YCHD. Results: Underfilling was the most common problem in preanalytical observation. This problem for citrate tubes was most frequently found in Vacusera® tubes. The possible cause of underfilling is the expiration date, which was close to the date of the experiment. There was a significant difference in PT results (p <0.0001) in the Vacusera® citrate tube. All tube brands had no significant differences in all clinical chemistry, hormonal, and hematological parameters (P>0.05). After 24 hours at room temperature, the results of hematological parameters showed statistically significant differences in MCV and hematocrit parameters, while other parameters were still within the CV limit of the instrument. Conclusion: There were no significant differences in the results of clinical chemistry, hormonal, and hematological parameters for all tube brands. A hematology parameters test should be done immediately as the delay may affect the MCV and hematocrit parameters.

Fluorescence In Situ Hybridization (FISH) for Detection of c-MYC Amplifications in AML Patients: A Potential Marker for Risk Stratification

Abstract Background c-MYC is a transcription factor (located on 8q24) which regulates genes involved in proliferation, cell growth, differentiation, and apoptosis. The c-MYC amplification is related to a rapid proliferation of the affected cell clone in patients with myeloid neoplasms. Gene amplification (amp) is one of the basic mechanisms connected with over-expression of oncogenes. c-MYC is routinely analyzed in mature lymphoid neoplasms especially in Burkitt’s Lymphoma while not assessed for acute myeloid leukemia (AML).Despite the fact the c- MYC oncogene is commonly over-expressed in AML it has remained an elusive target. Routine assessment for c- MYC in AML may pave the way for targeting the c-MYC in treatment of AML patients. Several experiments are ongoing investigating both direct and indirect c-MYC targeting therapies and further establishing cMYC as an oncogene of central importance to AML pathogenesis and treatment. Aim of the Work In this study we aimed to define the prognostic value of c-MYC amplification in adult AML patients, and to relate it to other prognostic factors in AML. Patients and Methods The study was a randomized control study, conducted at the Clinical Pathology Department, Ain-Shams University Hospitals during the period from January 2019 till June 2020. It was carried on 50 adult AML patients (30 newly diagnosed and 20 on Therapy) and 10 healthy age-matched volunteers as control. All subjects underwent full history taking, thorough clinical examination, laboratory investigations including; CBC, Examination of Leishman’s stained peripheral blood films, Bone marrow aspiration and examination of Leishman’s stained bone marrow smears, Cytochemical studies using Myeloperoxidase stain, Immunophenotyping of bone marrow or peripheral blood samples and Fluorescence in situ hybridization analysis (FISH). Results we had 9 patients with c-MYC amplification (6 in the newly diagnosed group and 3 in the on therapy group). So the percentage of c-MYC amplification in our patients was 9/50 (18%). To further elucidate the effect of c-MYC amplification on our patients’ clinical, laboratory and prognostic factors, an analytical comparison was done between negative and positive patients for cMYC amplification as regard-studied parameters in each group of patients. Both group of patients showed a statistical significant association between c-MYC amplification and outcome where the majority of patients with positive c-MYC amplification achieved incomplete remission. In addition, c-MYC amplification showed a statically significant association with Trisomy 8 in both patients’ groups and with 17p deletion and hepatosplenomegaly in on therapy group. Univariate logistic regression analysis for the association between c-MYC amplification and outcome showed statistically significant association between c-MYC amplification and incomplete remission in newly diagnosed AML patients with P-value = 0.013 and odds ratio (95% CI) of 14.0. Conclusion Our study has shown that c-MYC amplification is associated with adverse prognosis, poor treatment outcome and it might be involved in the process of AML progression. Targeting cMYC may be of value in the development of new-targeted AML therapeutics.

Factor V Leiden (FVL), Prothrombin G20210A and MTHFR C677T Mutations among Patients of Budd-Chiari Syndrome

Abstract Background Many studies ask whether screening Factor V Leiden (FVL), Prothrombin G20210A and Methyl Tetra Hydro Folate Reductase (MTHFR) C677T Mutations should be routinely performed in patients with Budd–Chiari syndrome (BCS). However, the prevalence of these mutations in such patients is substantially variable. This is the first study in clinical pathology department in Ain Shams University to evaluate the importance of this mutational screening in patients with Budd-Chiari syndrome to detect occult genetic susceptibility to thrombophilia. Aim of the Work This study aims to detect Factor V Leiden (FVL), Prothrombin G20210A and MTHFR C677T Mutations among Patients with Budd-Chiari Syndrome. Patients and Methods This study was a cross sectional study conducted on 35 newly of Budd Chiari syndrome patients who were attending tropical department at Ain Shams University Hospitals during the period from April 2020 to January 2021. Informed consent was obtained from all patients in the study. Results results of our study revealed that JAK2 mutation it was detected in 17.1% of patients (6 patients), While PGM was detected in 2.9% of our patients. Furthermore Factor V Leiden Mutation was detected in 65.7%of patients and MTHFR mutation was detected in 48.6% of patients. The present study found no significant statistical difference between patients with Factor V Leiden Mutation & patients with normal Factor V Leiden regarding CBC parameters, levels of ANA, ACL IgM, ACL IgG, PC Deficiency, PS Deficiency and AT III Deficiency. In addition, our study revealed that there was no effect of MTHFR Mutation & JAK2 Mutation on levels of ANA, ACL IgM, ACL IgG, PC Deficiency, PS Deficiency and AT III Deficiency. Also there was no effect for MTHFR Mutation & JAK2 Mutation on outcome of patients, survival and response to treatment. Conclusion BCS is associated usually with genetic thrombophilia. Most commonly Factor V Leiden Mutation in 65.7%, followed by MTHFR mutation in 48.6% of our patients. Although, only Factor V Leiden Mutation has an effect on patients Outcome, Survival and Response to treatment through its effect on CBC parameters, levels of ANA, ACL IgM, ACL IgG, PC Deficiency, PS Deficiency and AT III Deficiency. Otherwise, JAK2 Mutation have significant effect on hemopoiesis.

Quantitative Stool Culture of Candida in Autism Spectrum Disorder Patients

Abstract Background Autism spectrum disorder (ASD) is a category of neurodevelopmental disorders characterized by social and communication impairment and restricted or repetitive behaviors. Aim of the Work to estimate the quantity of Candida in autism spectrum disorder (ASD) patients compared to normal children and to find the association between Candida colony count and severity of ASD. Patients and Methods The study was carried out on 40 children, diagnosed as autism and 40 normal children as control group recruited from Pediatrics and adolescent Psychiatric Clinic at Ain Shams University Pediatrics Hospitals and Main Microbiology lab in Clinical Pathology Department of Ain Shams University. The study was conducted on a wide age group ranging from 3 to 14 years, (mean age of 6.30±2.40 years), Stool sample was collected from each patient in a sterile container, cultured on Sabouraud Dextrose Agar (SDA) and colony count was determined. Identification of isolated Candida species was done using chromogenic media. Results The study revealed statistically significant difference in Candida isolation rate among patient and control group with p-value (p = 0.006). Candida species were isolated from 23 patients (57.5%) and from 10 child in control group (25%), But detected that stool count was nonsignificant in severe autistic group compared to mild to moderate autistic group with p-value more than 0.05 NS. There were 14 patients (60.9%) with C. Albicans; 2 patients (8.7%) C. Glabrata; 6 patients (26.1%) C.Krusie and one patient (4.3%) C. Utilis, among candida species. Conclusion ASD is more common among male than female; ratio about 4:1. The high rate of intestinal colonization by Candida species in ASD children may represent a part of syndrome related to immune system disorders that may play a role in the pathogenesis of ASD. C. albicans was the predominant isolate than other non-Candida albicans species in ASD. The degree of candida colonization represented as quantitative colony count is not related to the severity of CARS among ASD patients.

Analysis of Factors That Cause the Target of Laboratory Critical Value Report Has Not Been Reached: Study at the Clinical Pathology Department at Haji General Hospital Surabaya

Background: A laboratory critical value is a laboratory test result represents a pathophysiologic state at such variance with normal which threaten patient’s life if a medical action is not taken immediately and for which an effective action is possible. The reporting of laboratory critical value at the Clinical Pathology Department at Haji General Hospital Surabaya for the January-July 2020 period had not reached 100%. Objectives: This study analyzed the factors cause the target of laboratory critical value report has not been reached at the Clinical Pathology Department at Haji General Hospital Surabaya based on psychological and organizational factors. Methods: This study was a descriptive observational study with cross-sectional approach. The samples were 23 people in total sampling. The data were collected by questionnaire that had been tested for validity and reliability. Results: The results showed that 73.9% of respondents had high motivation, 91.3% of respondents had good perceptions, 82.6% of respondents had type B personality, 69.6% of respondents had good learning, 78.3% respondents had low work stress, 52.2% of respondents stated that leadership was quite good, 95.7% of respondents said moderate rewards, 60.9% of respondents stated that the work design was according to the profession, 87.0% of the organizational structure was effective. Conclusion: Factors that caused target of laboratory critical value report has not been reached at the Clinical Pathology Department at Haji General Hospital Surabaya for the January-July 2020 period, based on the 80/20 Pareto Principle are motivation, learning, leadership, and job design. Health analysts should increase self-motivation, make experiences as learning, leaders as co-workers, work well willingly and the Clinical Pathology Department to reward the best employees, conduct evaluations, provide understanding and explanation in decision making, regulate the rotation of the division of tasks.

EDTA related Pseudothrombocytopenia (PTCP) Incidence and Resolution of Clinically Threatened Artifact

Background: EDTA related pseudothrombocytopenia (PTCP) is immune mediated, time and temperature dependent spurious low platelet counts commonly observed in laboratory practices, as an in vitro phenomenon. Microscopic evaluation of every sample is not possible due to immense workload hence PTCP is overlooked mostly due to immense work load .Vortex mixer is practically feasible tool often used in laboratories to obtain the accurate platelet counts of the suspected samples by using same EDTA anticoagulated samples. Aim: To evaluate the frequency of EDTA related pseudothrombocytopenia in OPD of tertiary care centre and analyze the vortexing for resolution of PTCP. Methodology: This was case control study conducted in clinical pathology department of JPMC in collaboration with ICU from November 2022 to January 2023. Total 3000 samples of CBC were analyzed. 150 patients with platelet count less than 150µl , with abnormal histograms and platelet clumps on smear were enrolled as cases of PTCP. 150 healthy individuals with match age and gender recruited as controls. Sample was subjected to vortex for 2 min at 3000rpm Samples were reanalyze ,clumps dissolution recheck by smear. Results : Prevalence of isolated thrombocytopenia was 33%, with 15% cases of EDTA related PTCP, platelet clumps seen in all PTCP cases on smear, 87% clumps were resolved completely with significantly increased platelet counts post vortexing with p value <0.05. Conclusion: Vortexing has proved as a promising, rapid and a cost effective method to deal with PTCP, save patient from needless investigations and invasive procedures. Keywords: Thrombocytopenia, Pseudothrombocytopenia, Ethylene diamine tetra acetic acid, Vortex, Platelet histograms,

Abstract 5442: SlideQC: An AI-based tool for automated quality control of whole-slide digital pathology images

Abstract Introduction: Artifacts are often introduced during tissue collection and processing, slide preparation, and/or when generating whole slide images (WSI). The presence of artifacts has a negative impact on the digital pathology workflow as artifacts may hinder diagnostic reporting and can lead to false positive and false negative results when using image analysis algorithms or computer-aided diagnosis systems. Manual quality control of WSI is a time-consuming procedure and therefore automated quality control tools, which report and exclude artifacts, are highly desirable to streamline digital pathology workflows. To automate the quality control step, we developed SlideQC, an AI-based quality control tool that automatically detects, reports, and outlines artifacts such as air bubbles, dust/debris, folds, out-of-focus,and pen marks, in both research and clinical workflows. Methods: SlideQC was trained with a DenseNet-based network using 1984 annotations for artifacts including air bubbles, dust/debris, folds, out-of-focus, and pen markers, across 254 Haematoxylin and Eosin (H&E) stained WSI from more than 9 tissue types. A set of 2048 annotations from synthetically generated out-of-focus images was added to supplement the training data. The performance of the SlideQC was evaluated on an external test cohort of 49 WSI H&E images sourced from the open-source database ‘HistoQCRepo’, across 375 annotations (tissue and artifact), and compared with the performance of HistoQC, an open-source quality control tool for digital pathology slides. Results: On the external test cohort, SlideQC showed high sensitivity, specificity, and F1-score with average values of 0.93, 0.99, and 0.93, across the five artifact types. In the same cohort, HistoQC attained an average sensitivity, specificity, and F1-score of 0.65, 0.79, and 0.54, respectively. Conclusions: SlideQC achieved high sensitivity, specificity, and F1-score on an external test cohort. SlideQC can add efficiency gains to a workflow by performing quality control on 100% of slides rather than the currently manually performed on only a subset of the slides in clinical pathology departments. SlideQC can allowthe triaging and alerting of slides containing a high level of artifact within a digital pathology workflow. The tool can also be used to exclude the artifact region from downstream analysis by subsequent image analysis algorithms. Citation Format: Daniela Rodrigues, Stefan Reinhard, Therese Waldburger, Daniel Martin, Suzana Couto, Inti Zlobec, Peter Caie, Erik Burlingame. SlideQC: An AI-based tool for automated quality control of whole-slide digital pathology images. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5442.

A New Artificial Intelligence Approach Using Extreme Learning Machine as the Potentially Effective Model to Predict and Analyze the Diagnosis of Anemia

The procedure to diagnose anemia is time-consuming and resource-intensive due to the existence of a multitude of symptoms that can be felt physically or seen visually. Anemia also has several forms, which can be distinguished based on several characteristics. It is possible to diagnose anemia through a quick, affordable, and easily accessible laboratory test known as the complete blood count (CBC), but the method cannot directly identify different kinds of anemia. Therefore, further tests are required to establish a gold standard for the type of anemia in a patient. These tests are uncommon in settings that offer healthcare on a smaller scale because they require expensive equipment. Moreover, it is also difficult to discern between beta thalassemia trait (BTT), iron deficiency anemia (IDA), hemoglobin E (HbE), and combination anemias despite the presence of multiple red blood cell (RBC) formulas and indices with differing optimal cutoff values. This is due to the existence of several varieties of anemia in individuals, making it difficult to distinguish between BTT, IDA, HbE, and combinations. Therefore, a more precise and automated prediction model is proposed to distinguish these four types to accelerate the identification process for doctors. Historical data were retrieved from the Laboratory of the Department of Clinical Pathology and Laboratory Medicine, Faculty of Medicine, Public Health, and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia for this purpose. Furthermore, the model was developed using the algorithm for the extreme learning machine (ELM). This was followed by the measurement of the performance using the confusion matrix and 190 data representing the four classes, and the results showed 99.21% accuracy, 98.44% sensitivity, 99.30% precision, and an F1 score of 98.84%.

The study of long noncoding RNA SNHG5 and PANDAR genes expression in newly diagnosed egyptian adult acute myeloid leukemia patients

BACKGROUND: Due to their impact on crucial steps in hematopoiesis, long noncoding RNAs (lncRNAs) deregulation potentially accelerates the growth and development of blood cancers like acute myeloid leukemia (AML). The study aimed to look into different expression patterns, prognostic value, and clinical importance of lncRNA small nucleolar RNA host gene 5 (SNHG5) and promoter of cyclin-dependent kinase inhibitor 1A antisense DNA damage-activated RNA (PANDAR) genes in Egyptian adult patients with AML. SUBJECTS AND METHODS: The case–control study was conducted between 2019 and 2022 at the Clinical Pathology Department at the National Cancer Institute, Cairo University, Egypt. The study involved 80 recently diagnosed patients with AML and 20 healthy controls. Real-time quantitative reverse transcription polymerase chain reaction was used to assess the levels of expression of SNHG5 and PANDAR genes. RESULTS: In comparison to healthy controls, there was a significantly higher SNHG5 gene expression (P = 0.026) and PANDAR expression (P < 0.001) in patients’ bone marrow samples. The study of the correlations revealed a significant positive association between SNHG5 and PANDAR genes in AML patients. The overall survival (OS) was significantly better in the low SNHG5 gene expression group than in the high SNHG5 gene expression group. No significant difference was detected regarding the disease-free survival (DFS) between patients with low expression and high expression of the SNHG5 gene. No significant variation between high PANDAR gene and low PANDAR gene expression regarding OS and DFS. CONCLUSION: SNHG5 and PANDAR may have a pathogenic role in AML, and their overexpression might be considered a marker for diagnosis in AML patients in Egypt. SNHG5 expression can be used as a predictor for OS, while PANDAR expression cannot be used as a predictor for OS or DFS in patients.

A case of nodular enteritis due to Chaunocephalus ferox in a white stork (Ciconia ciconia) in Bulgaria

The present report describes a case of chaunocephalosis due to Chaunocephalus ferox in a white stork (Ciconia ciconia) in Bulgaria. On July 2017, a malnourished and debilitated adult white stork was found in a field near Stara Zagora city. The bird was caught and transported to the Wildlife Rescue Center of Green Balkans organisation, with history of weakness, severe watery diarrhoea, loss of appetite, and cachexia. The stork died several hours after admission. The corpse was referred to the Department of General and Clinical Pathology, Faculty of Veterinary Medicine, Trakia University where post-mortem examination was performed. The necropsy revealed the presence of multifocal to coalescing mural intestinal nodular circumscribed lesions affecting all portions of the intestinal tract, most prominent and numerous in the jejuno-ileal segment. When sectioned, the nodules contained either single, or 2 to 3 small flattened tadpole-like trematodes which were identified as Ch. ferox. Histologically, a total traumatic destruction with ulceration and necroses of intestinal tissue layers were found at the site of fluke penetration. The trematodes also formed a marked granulomatous inflammatory reaction with inflammatory cellular infiltrate, consisting mainly of heterophilic and pseudoeosinophilic leukocytes, lymphocytes and histiocytes. In the area of infection, the intestinal villi and mucosa were totally eroded, and the surrounding mucosal crypts were distended by necrotic detritus.

Clinical utility of haematological inflammatory biomarkers in predicting 30-day mortality in hospitalised adult patients with COVID-19.

Coronavirus disease 2019 (COVID-19) is a multisystem disease affecting respiratory, cardiovascular, gastrointestinal, neurological, immunological and haematological systems. The most important indices that have been studied are platelet (PLT) indices in addition to the PLT count and red blood cell distribution width (RDW). This retrospective study included 95 patients with COVID-19 and was conducted at the Hospital Isolation, Scientific and Medical Research Centre and Clinical Pathology Department at Zagazig University Hospitals, Egypt over 6 months from March to August 2021. All patients on admission had a full blood count, which included white blood cell (WBC) count, haemoglobin, RDW, PLT count and its indices in addition to PLT-to-WBC ratio (PWR) and PLT-to-lymphocyte ratio (PLR), which were calculated for all the study patients. There were significant linear correlations for higher levels of the PLR, PWR and RDW and mortality rate (p=0.03, p < 0.001 and p < 0.001 respectively). Moreover, on multivariable analysis the RDW, PLT count and PWR levels were independent prognostic predictors for mortality with a hazard ratio [HR] of 1.25 (95% confidence interval [CI] 1.09-1.44, p=0.002), 1.00 (95% CI 0.99-1.00, p=0.03) and 2.3 (95% CI 1.21-4.48, p=0.01) respectively. The RDW and PLT indices are accessible predictors that can be valuable prognostic factors for survival assessment and risk stratification of COVID-19.

Eosinopenia As a Marker of Sepsis in Intensive Care Unit Patients

Background: Sepsis is one of the most common causes of mortality and morbidity in the intensive care unit (ICU). Objective: This study was done to evaluate eosinopenia as a marker of sepsis in intensive care unit patients. Materials and Methods: This cross sectional study was carried out in the Department of Clinical Pathology, Bangabandhu Sheikh Mujib Medical University, Dhaka . AEC was done by automated cell counter and rechecked manually microscopically. Observations and Results: In this study 74 suspected case of sepsis were enrolled from intensive care unit, BSMMU, Dhaka. To evaluate the usefulness of AEC for predicting significant sepsis the area under the ROC curve was analyzed. The area under ROC curve was 0.765 in current study. At the cut off valve of AEC &lt; 40 cell/cu mm, the sensitivity and specificity of AEC for diagnosis of sepsis was 72.5% and 61.8% respectively. Conclusion: This present data revealed that decreased absolute eosinophil count was significantly associated with sepsis. So eosinopenia may be a reliable marker for early diagnosis of sepsis. Sir Salimullah Med Coll J 2022; 30: 175-179