CDCA8, a mitosis-related gene, as a prospective pan-cancer biomarker: implications for survival prognosis and oncogenic immunology.

BACKGROUND: Human cell division cycle-associated protein 8 (CDCA8), a critical regulator of mitosis, has been identified as a prospective prognostic biomarker in several cancer types, including breast, colon, and lung cancers. This study analyzed the diagnostic/prognostic potential and clinical implications of CDCA8 across diverse cancers. METHODS: Bioinformatics and molecular experiments. RESULTS: Analyzing TCGA data via TIMER2 and GEPIA2 databases revealed significant up-regulation of CDCA8 in 23 cancer types compared to normal tissues. Prognostically, elevated CDCA8 expression correlated with poorer overall survival in KIRC, LUAD, and SKCM, emphasizing its potential as a prognostic marker. UALCAN analysis demonstrated CDCA8 up-regulation based on clinical variables, such as cancer stage, race, and gender, in these cancers. Epigenetic exploration indicated reduced CDCA8 promoter methylation levels in Kidney Renal Clear Cell Carcinoma (KIRC), Lung Adenocarcinoma (LUAD), and Skin Cutaneous Melanoma (SKCM) tissues compared to normal controls. Promoter methylation and mutational analyses showcased a hypomethylation and low mutation rate for CDCA8 in these cancers. Correlation analysis revealed positive associations between CDCA8 expression and infiltrating immune cells, particularly CD8+ and CD4+ T cells. Protein-protein interaction (PPI) network analysis unveiled key interacting proteins, while gene enrichment analysis highlighted their involvement in crucial cellular processes and pathways. Additionally, exploration of CDCA8-associated drugs through DrugBank presented potential therapeutic options for KIRC, LUAD, and SKCM. In vitro validation using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) confirmed elevated CDCA8 expression in LUAD cell lines (A549 and H1299) compared to control cell lines (Beas-2B and NL-20). CONCLUSION: This study provides concise insights into CDCA8's multifaceted role in KIRC, LUAD, and SKCM, covering expression patterns, diagnostic and prognostic relevance, epigenetic regulation, mutational landscape, immune infiltration, and therapeutic implications.

Reporting Practices of Serum Protein Electrophoresis in Pakistan - a Multicenter Survey.

BACKGROUND: Serum Protein Electrophoresis (SPE) is crucial for the diagnosis and follow-up of monoclonal gammopathy (MG), as it helps to separate and identify these paraproteins. Currently, Pakistan lacks standardized guidelines for SPE reporting and analytical performance. This survey aims to analyze reporting variations from Consultant Chemical Pathologists in Pakistani laboratories. METHODS: This cross-sectional survey was conducted by the section of Chemical Pathology, Department of Pathology and Laboratory Medicine, at Aga Khan University Hospital, Karachi. A previously validated and published tool was used with some modifications to assess analytical techniques, reporting patterns, and interpretations provided with SPE by different laboratories. Frequency and percentages were calculated for each response and descriptive results were also evaluated. Differences between laboratories were also assessed qualitatively. RESULTS: Out of the eight laboratories contacted, seven participated in the survey, yielding a response rate of 87.5%. Immunofixation Electrophoresis (IFE) was used by all labs for serum immunotyping. All labs reported a new small abnormal band in patients with no known monoclonal gammopathy or with a known M-protein. Variations were found in terminologies used to label paraprotein, terminologies used to report normal and pathological SPE patterns, electrophoretic technique, methods for quantifying paraprotein in the gamma region on SPE and for albumin quantification. Similarly, the number of decimal places reported, reporting of multiple monoclonal proteins and small paraprotein in the beta region or monoclonal proteins less than 1 g/L, approach for screening, number of fractions reported in gamma region and reporting of interferences were also not standardized and var-iations were noticed. CONCLUSIONS: Our survey highlighted variations in practices of SPE reporting. These differences in laboratory practices could result in inconsistent test results, which could adversely affect patient care.

Rare serosal cystic gastric gastrointestinal stromal tumor with extensive intestinal metaplasia in an adherent gastric mucosa; a case report in a 65-year-old male.

Gastrointestinal (GI) intestinal stromal tumors account for 60% of mesenchymal GI tract tumors commonly located in the stomach and small intestine, predominantly solid tumors that rarely undergo cystic degeneration. A 65-year-old patient with increasing upper abdominal swelling and a computed tomography scan abdomen showed a large unilocular 17 × 16 × 15 cm lesion. A colossal cystic swelling in the lesser omentum, anterior to the stomach, was found upon exploration. Histopathological examination showed a spindle cell tumor turned out to be CD117 positive and S100 negative on immunostains. The tumor was moderate risk gastric gastrointestinal intestinal stromal tumor (GIST) based on the site; Stomach, Size >10 cm; Mitosis <5/5 mm2 according to risk assessment of GIST, 2006. GISTs are predominantly solid tumors and rarely undergo cystic transformation. The primary differential diagnoses of spindle cell neoplasm are GISTs, Leiomyoma, Leiomyosarcoma and Schwannoma. These spindle cell neoplasms are differentiated by applying a panel of Immunohistochemical stains, CD117, SMA and S100.

Clinical and Biochemical Characteristics of COVID-19 Patients During the Delta-Omicron Wave with Risk Assessment of Adverse Outcomes.

OBJECTIVE: To compare clinical and biochemical characteristics of hospitalised COVID-19 patients and risk assessment of disease outcomes Study Design: Descriptive study. Place and Duration of the Study: Department of Pathology, Dow International Medical College and Sindh Infectious Diseases Hospital and Research Centre, from January to March 2022 Methodology: SARS CoV-2 PCR-positive hospitalised patients were enrolled. Delta or omicron variants infected patients were followed till the last recorded event of hospitalisation. After a detailed history, clinical and biochemical profiles were recorded during the hospitalisation. Length of hospitalisation, ICU admission and in-hospital mortality were taken as outcomes and odd ratios were calculated. RESULTS: During the study period, omicron was the predominant SARS CoV-2 variant. Omicron-infected patient were older (67 vs. 62 years) and had a significantly shorter duration between appearance of symptoms and hospitalisation (5 vs. 8 days), when compared with the delta patients. Median values of LDH, ferritin and TLC were significantly higher in delta patients (p<0.05). Delta infected patients have a 3.9 times more risk of prolonged hospital stay. In patients with increased TLC, the risk of prolonged hospitalisation and ICU admission was found 16% and 23%, respectively. However, the aOR for ICU admission and in- Hospital mortality were not found significant for the delta and omicron-infected patients. CONCLUSION: The clinical course and biochemical profiles are diverse in delta and omicron patients. Hospitalised patients with omicron infection exhibit shorter stays. High values of TLC are found associated with an increased risk of longer hospital stay and ICU admissions. KEY WORDS: COVID-19, Delta variant, Omicron variant, Hospitalised patients, Outcomes, In-hospital mortality, Biochemical markers, Clinical severity.

Comparison Of Hepatitis C Antibody Assays And Evaluation Of Agreement Between Hepatitis C Antigen And PCR Results.

BACKGROUND: Hepatitis C is associated with a wide range of health repercussions. Pakistan is one of the highly prevalent countries of Hepatitis C Virus (HCV) infection. The availability of cost-effective, robust, and reliable screening and diagnostic tests for hepatitis C is important to address the disease burden. Standardization of screening and diagnostic assays in clinical laboratories is crucial for achieving big goals. Objectives of this study are to correlate the results of two different HCV antibody (HCV Ab) assays and to examine the correlation of HCV core antigen (HCV c Ag) results with HCV PCR for HCV infection diagnosis. METHODS: This descriptive cross-sectional study was carried out from November to December 2020 at Dow University of Health Sciences. Total number of 40 HCV Ab samples were analysed by both chemiluminescence (CMIA) and electrochemiluminescence (ECLIA) immunoassays. Tests for HCV RNA PCR and HCV c Ag were performed on all samples. Results of screening and diagnostic assays were correlated and agreements were examined. Statistical analysis for agreement was carried out by using R software version 3.6.3 through AC1 Gwetz Statistic. The study was approved by the institutional ethical review committee. RESULTS: An agreement of 0.73 and 0.95 was found between two different HCV Ab immunoassays and HCV c Ag and HCV PCR, respectively. CONCLUSIONS: We found a good correlation between CMIA and ECLIA for HCV Ab. An excellent correlation was found between HCV c Ag and HCV PCR. Based on our study findings, HCV c Ag is a candidate test for the diagnosis of active HCV infection.

Development of a virtual classroom for pre-analytical phase of laboratory medicine for undergraduate medical students using the Delphi technique.

BACKGROUND: Amongst the pre-analytical, analytical, and post-analytical phase of laboratory testing, pre-analytical phase is the most error-prone. Knowledge gaps in understanding of pre-analytical factors are identified in the clinical years amongst undergraduate students due to lack of formal teaching modules on the pre-analytical phase. This study was conducted to seek experts' consensus in Clinical Chemistry on learning objectives and contents using the Delphi technique with an aim to develop an asynchronous virtual classroom for teaching pre-analytical factors of laboratory testing. METHODS: A mixed method study was conducted at the Aga Khan University. A questionnaire comprising of 16 learning objectives and their associated triggers was developed on Google Docs for developing the case vignettes. A four-point Likert Scale, which included strongly agree, agree, disagree and strongly disagree, was utilized for the learning objectives. An open-ended question was included for experts to suggest new items for inclusion. A cut off of at least 75% agreement was set to establish consensus on each item. A total of 17 Chemical Pathology faculty from 13 institutions across Pakistan were invited to participate in the first round of Delphi. Similar method of response was used in round two to establish consensus on the newly identified items suggested by the faculty in round 1. Later, the agreed-upon objectives and triggers were used to develop interactive scenarios over Moodle to concurrently test and teach medical students in a nonchalant manner. RESULTS: A total of 17 responses were received in Round 1 of the Delphi process (response rate = 100%), while 12 responses were received in Round 2 (response rate = 71%). In round 1, all 16 learning objectives reached the required consensus (≥ 75%) with no additional learning objectives suggested by the experts. Out of 75 triggers in round 1, 61 (81.3%) reached the consensus to be included while 39 were additionally suggested. In 2nd round, 17 out of 39 newly suggested triggers met the desired consensus. 14 triggers did not reach the consensus after two rounds, and were therefore eliminated. The virtual classroom developed using the agreed-upon learning objectives and triggers consisted of 20 items with a total score of 31 marks. The questions included multiple choice questions, fill in the blanks, drag and drop sequences and read-and-answer comprehensions. Specific learning points were included after each item and graphs and pictures were included for a vibrant experience. CONCLUSION: We developed an effective and interactive virtual session with expert consensus on the pre-analytical phase of laboratory testing for undergraduate medical students which can be used for medical technologist, graduate students and fellows in Chemical Pathology.

Relationship Between Psychosocial Factors and Glucose Control in Adults With Type 2 Diabetes.

OBJECTIVES: Despite significant advances in medical therapy and unrestricted access to health care, >50% patients with type 2 diabetes (T2D) cannot maintain their blood glucose target levels. This cross-sectional study investigated the association between psychosocial behaviour and diabetes management in Newfoundland and Labrador, where the prevalence of T2D is the highest in Canada. METHODS: Data were collected from 165 adult T2D patients. Four sets of self-administered standardized questionnaires, a study-specific data form and electronic health records were utilized to obtain psychosocial information, patient characteristics and glycated hemoglobin (A1C) levels. RESULTS: The group of participants with emotional burnout due to diabetes-related stress showed poor glycemic control (89.4%) compared to the group with low stress (55.6%). The group with higher stress appraised T2D negatively (correlation coefficient r=0.719, and p<0.01), and had a tendency to use emotion-oriented coping (r=0.542, p<0.01) and had a poor perception of autonomous supportiveness (r=-0.300, p<0.01). A path model showed that stress, appraisal and coping explained 7.4% of the variance in A1C. Appraisal plays the role of mediator and explained 5.8% of the variance in A1C. CONCLUSIONS: A high prevalence of poor glycemic control was found in participants with a body mass index of ≥35. Participants with higher stress had a negative appraisal of T2D. The highly stressed group tended to use emotion-oriented coping and have a poor perception of autonomous supportiveness.

Novel psychoactive substances: overdose of 3-fluorophenmetrazine (3-FPM) and etizolam in a 33-year-old man.

Though illegal in the UK, in many countries novel psychoactive substances are quasi-legal synthetic compounds that are widely available online under the guise of research chemicals. These substances are relatively cheap and are often undetectable in standard drug screens. Nearly 200 such compounds are introduced yearly, and little is usually known about their metabolism or physiological effects. Consequently, managing patients in overdose situations on largely unknown substances usually involves supportive care, however anticipating and managing atypical side effects are challenging in the absence of knowledge of these compounds. In this report, we discuss our encounter with a 33-year-old unconscious man presenting with coingestion of a novel stimulant 3-fluorophenmetrazine with a rarely used benzodiazepine etizolam. This patient developed seizure-like activity and delayed widespread T-wave inversions, both of which ultimately resolved without sequelae.

Estimation of uncertainty measurement - A prerequisite of ISO1589 accreditation for clinical laboratories.

OBJECTIVE: To estimate relative expanded uncertainty measurement of routine clinical chemistry analytes for international organisation for standardisation 15189 accreditation. METHODS: This cross-sectional study was conducted at Dow International Medical College, Karachi, from September 2013 to May 2014. During the process of international organisation for standardisation 15189 accreditation, measurement uncertainty was estimated for 13 clinical chemistry analytes using top-down approach. Relative combined uncertainty of each analyte was calculated by combining uncertainties of imprecision, bias and calibrators. Results of estimated imprecision, bias and expanded uncertainties were observed for allowable imprecision, bias and total analytical error for the respective analyte.. RESULTS: Uncertainties of imprecision were found within acceptable limits for all analytes except total protein (2.4% vs. 1.3%). Uncertainties of bias of all analytes were found within allowable limits. Relative expended uncertainties of all analytes were found acceptable except total protein (4.7%vs 3.63%). CONCLUSIONS: The approach used to estimate the measurement uncertainty may be found simple and feasible by clinical laboratories interested in getting the relevant accreditation.

Application of Sigma Metrics Analysis for the Assessment and Modification of Quality Control Program in the Clinical Chemistry Laboratory of a Tertiary Care Hospital.

Sigma is a metric that quantifies the performance of a process as a rate of Defects-Per-Million opportunities. In clinical laboratories, sigma metric analysis is used to assess the performance of laboratory process system. Sigma metric is also used as a quality management strategy for a laboratory process to improve the quality by addressing the errors after identification. The aim of this study is to evaluate the errors in quality control of analytical phase of laboratory system by sigma metric. For this purpose sigma metric analysis was done for analytes using the internal and external quality control as quality indicators. Results of sigma metric analysis were used to identify the gaps and need for modification in the strategy of laboratory quality control procedure. Sigma metric was calculated for quality control program of ten clinical chemistry analytes including glucose, chloride, cholesterol, triglyceride, HDL, albumin, direct bilirubin, total bilirubin, protein and creatinine, at two control levels. To calculate the sigma metric imprecision and bias was calculated with internal and external quality control data, respectively. The minimum acceptable performance was considered as 3 sigma. Westgard sigma rules were applied to customize the quality control procedure. Sigma level was found acceptable (≥3) for glucose (L2), cholesterol, triglyceride, HDL, direct bilirubin and creatinine at both levels of control. For rest of the analytes sigma metric was found <3. The lowest value for sigma was found for chloride (1.1) at L2. The highest value of sigma was found for creatinine (10.1) at L3. HDL was found with the highest sigma values at both control levels (8.8 and 8.0 at L2 and L3, respectively). We conclude that analytes with the sigma value <3 are required strict monitoring and modification in quality control procedure. In this study application of sigma rules provided us the practical solution for improved and focused design of QC procedure.

Frequency of Thyroid Peroxidase Antibody and its Association with Miscarriages Among Pregnant Women.

OBJECTIVE: To measure the frequency of thyroid peroxidase antibody (TPO Ab) among clinically euthyroid pregnant women during first trimester and determine its association with pregnancy outcome as miscarriage or live birth by estimating the relative risk. STUDY DESIGN: Cohort study. PLACE AND DURATION OF STUDY: Section of Chemical Pathology, Department of Pathology and Laboratory Medicine and the Gynaecology and Obstetric outpatient clinics of the Aga Khan University Hospital, Karachi, from July to December 2012. METHODOLOGY: The study subjects comprised of apparently euthyroid pregnant women, who were tested for TPO Ab during first trimester of pregnancy and followed till pregnancy outcomes. Pregnancy outcome was noted and relative risk was determined. RESULTS: TPO Ab was found positive in 127 (13.5%) pregnant women from a cohort of 943 subjects. A2.03% increased risk of miscarriages was observed in TPO Ab positive subjects. CONCLUSION: There was a positive association of presence of TPO Ab with loss of pregnancy.

Agreement between serology and histology for detection of Helicobacter pylori infection.

OBJECTIVE: To determine the percentage agreement between serology and histology for detection of Helicobacter (H.) pylori infection. STUDY DESIGN: Cross-sectional analytical study. PLACE AND DURATION OF STUDY: Department of Pathology and Microbiology, The Aga Khan University and Hospital, Karachi, from January to December 2009. METHODOLOGY: Fifty subjects were selected by non-probability purposive sampling from laboratory data who had serological testing of H. pylori IgG antibody, prior to histological evaluation of endoscopic gastric or/and duodenal biopsies. Serological Quantification of H. pylori IgG was carried out with HpG screen ELISA kit (Genesis Diagnostics, UK), using an enzyme linked immunosorbent assay for detection of IgG antibodies against H. pylori. Manufacturer's recommended cutoff value was used and results were considered positive when greater than 7 U/ml. For histological diagnosis, an expert histopathologist characterized the presence of spiral bacteria in the mucosal layer or the surface of epithelial cells on microscopic examination, as a positive test. RESULTS: An agreement of 0.72 was found by Kappa statistics between serology and histopathology results and a good diagnostic accuracy (86%) of serological testing was observed for the diagnosis of H. pylori infection. CONCLUSION: A substantial agreement was found between serology and histopathology results to detect the H. pylori infection. Laboratory-based serologic testing using ELISA technology to detect IgG antibodies is inexpensive, noninvasive and convenient method to detect the H. pylori infection in primary care setting.

Raised 17-hydroxyprogesterone levels in congenital adrenal hyperplasia.

Congenital adrenal hyperplasia (CAH) refers to autosomal recessive diseases resulting from deficiency of enzymes involved in the production of cortisol by the adrenal glands. This study was designed to determine the frequency of suspected congenital adrenal hyperplasia patients by evaluating the laboratory data of blood 17-OHP. The study was conducted at Chemical Pathology Section of Department of Pathology at the Aga Khan University, Karachi. The basic demographic data of 2282 subjects was recorded, screened for blood 17-OHP levels from January 2007 to December 2010. A cutoff of ³ 4 ng/ml was considered as suggestive of CAH. The results showed 17-OHP levels ³ 4 ng/ml were found predominantly among infants (14.4%) and in females (18.2%).

Verification and Determination of the Thyroid Peroxidase Antibody Reference Interval: Insight Into the CLSI Guideline.

OBJECTIVE: To verify the upper reference limit and define the population-based reference interval (RI) for thyroid peroxidase antibody, according to Clinical and Laboratory Standards Institute (CLSI) guidelines. MATERIAL AND METHODS: We subjected serum samples from 146 subjects to thyroid-stimulating hormone (TSH) and TPO Ab analysis for verification and determination of RI. We verified an upper reference limit for thyroid peroxidase antibody (TPO Ab) and established an RI according to National Academy of Clinical Biochemistry (NACB) and CLSI guidelines. RESULTS: The manufacturer's suggested upper reference limit was not verified for use as a functional reference value. Rather, on defining the RI of TPO Ab by means of a full-scale study, we found a higher upper reference limit (13.8 vs 12.0). We calculated lower and upper reference limits of 3.3 and 13.8, with 90% confidence intervals (CIs) of 2.8 to 3.5 and 13.6 to 14.8, respectively. CONCLUSION: RIs provided by assay manufacturers are general data and may not always be appropriate for different locations and patient demographics. Verification of an RI is necessary before use in a given patient population.