Electrochemical biosensing of circulating MicroRNAs in endometrial Cancer.

A 90-day oral toxicity study of heat-treated Bifidobacterium longum CECT 7347 (ES1 HT) Postbiotic in rats.

Biochemistry is fundamental to medical education; however, traditional teaching may limit clinical integration and long-term retention. Although clinical simulation has emerged as a strategy to promote active and contextualized learning, its effectiveness in biochemistry teaching remains limited. This study evaluated the impact of clinical simulation on cognitive knowledge, technical skills, and communication among first-semester medical students compared with traditional practical instruction. A mixed-methods, quasi-experimental design was used with 404 students from the Central University of Ecuador, allocated to a simulation group (n = 205) or control group (n = 199). Both groups received identical theoretical instruction. Outcomes were assessed at baseline and post-intervention in four domains: theoretical knowledge, clinical interpretation, technical skills, and communication. Student satisfaction and perceptions were evaluated using surveys and qualitative analysis. Both groups improved after sessions; however, the simulation group demonstrated significantly greater gains across all domains (theoretical knowledge, clinical interpretation, technical skills, and communication). Students in the simulation-based group also achieved higher success rates in blood-sampling procedures, completed tasks in less time, and reported more positive emotions, reduced anxiety, increased confidence, and greater satisfaction. Simulation-based education appears to be an effective and feasible approach to enhanceknowledge, technical, communication skills, and student motivation in medical biochemistry teaching.

To help assess patient safety risks associated with ongoing differences in total calcium assays following bias shifts, we performed spike-recovery experiments within our clinical chemistry external quality assessment (EQA) schemes to evaluate bias and recovery against a reference measurement procedure (RMP). Aqueous calcium chloride dihydrate was spiked (0.30/0.60 mmol/L; 1.20/2.40 mg/dL) into pooled human serum. Bias was assessed against RMP-assigned targets and recovery against added spikes. Minimum analytical performance specifications (APS) for standard measurement uncertainty (1.36%) defined acceptable bias/recovery. Commercially available analytical principles across analytical platforms did not achieve adequate bias against RMP-assigned targets and recovery against added spikes for total calcium in 3 EQA specimens. Roche Cobas (5-nitro-5'-methyl-1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid [NM-BAPTA]), Roche Cobas Pro (NM-BAPTA), and Beckman AU (Arsenazo III) methods exhibited better bias and long-term stability. The Abbott Alinity (Arsenazo III) method exhibited notable negative bias (-2.33% to -3.10%), apparently coinciding with the introduction of Abbott Calcium2. Negative bias was present to a lesser extent in the Abbott Architect (Arsenazo III). The Siemens Atellica (cresolphthalein complexone) assay exhibited notable proportional over recovery (110.2% to 112.0%). There was some positive bias in the Siemens Atellica (Arsenazo III) and QuidelOrtho (dry slide/sensor) methods. Differences across analytical principles/platforms and difficulties achieving minimum APS for total calcium were shown within our EQA schemes, notably negative bias for Abbott Alinity and Architect (Arsenazo III), over-recovery for Siemens Atellica (cresolphthalein complexone) and positive bias for Siemens Atellica (Arsenazo III). Understanding the impact on clinical decision-making and patient outcomes is imperative for managing any patient safety risks.

In preclinical toxicity and efficacy studies, clinical pathology tests play a critical role in detecting pathological alterations, assessing drug effectiveness, and monitoring animal conditions. However, their application is frequently restricted by the limited sample volumes that can be collected from laboratory animals such as mice and young animals. To overcoming this limitation, dilution of samples may be utilized. In this study, we investigated the impact of pre-dilution by using a Toshiba analyzer and saline as the diluent, comparing analytical results from diluted and undiluted specimens in both mice and rats. Seventeen biochemical analytes were assessed in serum samples diluted 2.5-, 5-, 7.5-, and 10-fold, and these results were compared with those of undiluted samples. We evaluated mean values, bias, two standard deviations (2SD), and the standard deviation index (SDI). To ensure data obtained from diluted samples are valid and dependable, we applied allowable total error (TEa), defined as the maximum acceptable error for a test result to remain reliable and clinically relevant. The mean values from 2.5-fold diluted samples exhibited no significant difference compared to those from undiluted samples in mice and rats, except Cl, Na, ALB, and TBIL. Furthermore, findings from bias, 2SD, SDI, and TEa analyses indicated that a 2.5-fold sample dilution is suitable for clinical chemistry analysis of multiple analytes, except for Cl, Na, and TBIL in both species. Collectively, these results may serve as a reference for the suitability of diluted serum samples in clinical chemistry analysis. Further studies utilizing various analytical instruments, diluents, or animal species are recommended.

Bovine hemoplasmas, including Mycoplasma wenyonii (M. wenyonii) and Candidatus Mycoplasma haemobos (C. M. haemobos), are vector-borne pathogens that can cause significant clinical diseases in cattle. This case report from Türkiye describes the clinical findings, hematological analysis, and serum biochemistry results of a cow presenting with hemoglobinuria caused by C. M. haemobos. Blood samples from the infected animal were analyzed using hematological, biochemical, microscopic, and molecular methods. Multiplex polymerase chain reaction assay confirmed the infection, and microscopic analysis revealed a parasitemia rate of 6.99%. Clinical signs included hemoglobinuria, icterus, and anemia. The treatment included fluid therapy, antibiotic (oxytetracycline), and antianemic drugs. The cattle died 1 month post-treatment, reportedly due to a lower respiratory tract infection. This case report describes the diagnostic methods, clinical findings, and laboratory results. A widescale molecular survey of C. M. haemobos in cattle is warranted, with investigation of its pathogenicity and pathogenies.

Circulating microbial metabolites and the gut-prostate axis in prostate cancer: Implications for laboratory biomarkers and therapeutic response.

Medical laboratory technologists face challenges related to inconsistent formal education, insufficient post-qualification training opportunities, and limited oversight by a national accreditation body. The purpose of this study was to obtain consensus from field experts on the exit competencies essential for medical laboratory graduates for developing a competency-based curriculum for an MLT training program. Using the CDC/APHL competency framework developed by the Centers for Disease Control and Prevention (USA) and the Association of Public Health Laboratories, experts from related disciplines selected relevant competencies, followed by a Delphi survey to establish consensus. Responses were collected via a Likert scale, and consensus was defined as ≥ 80% agreement. Data was analyzed using SPSS. Ethical approval was obtained from the AKUH Ethical Review Committee. A total of 11 workshops were conducted in a flipped format to finalize 106 competencies from the source guideline and included in the Delphi questionnaire. Ninety-five competencies achieved ≥ 80% agreement in the 1st round, and three competencies reached agreement in the 2nd round. The competencies spanned core system-level and operational domains such as quality management systems, ethics, leadership, communication, safety, surveillance, and informatics, and were applied across laboratory disciplines, including microbiology, clinical chemistry, hematology and blood banking, bioinformatics, and research. This Delphi study identified 98 competencies for MLSs across 17 domains, providing the first structured, contextually grounded competency framework for a graduate program in MLS education in Pakistan.

Live biotherapeutic products are emerging as supports for host physiology and gastrointestinal health. However, despite their broad use and apparent safety, the in vivo toxicity of new probiotic strains must be rigorously evaluated before human application. Bacillus safensis NMCC-189, a spore-forming bacterium with promising probiotic potential, was assessed through a 14-day short-term repeated-dose tolerability study to confirm initial safety followed by sub-acute (28-day) oral toxicity study conducted in accordance with Organisation for Economic Co-operation and Development (OECD) test guideline 407. Male and female Balb/c mice were orally administered low (1 × 108CFU/mL) and high doses (1 × 1010CFU/mL) of B. safensis NMCC-189 corresponding to approximately 1 × 109CFU/kg/day 1 × 1011CFU/kg/day. Dose selection was based on established probiotic safety ranges and toxicological principles. Clinical signs, survival, body weight, food intake, haematology, serum biochemistry, organ weights, and histopathology were examined, alongside daily in-life monitoring, randomisation, and blinding. No morbidity, mortality, or treatment related clinical abnormalities were observed in either study phase. Body weight gain, food consumption, and behavioural patterns remained within normal physiological limits. Haematological and biochemical parameters showed no toxicologically relevant changes, and minor fluctuations remained within reference ranges. Gross and microscopic examination of major organs revealed no pathological changes. The no observed adverse effect level (NOAEL) was 1 × 1011CFU/kg/day. These findings suggest that B. safensis is well tolerated following oral administration and support its safety as a probiotic candidate. Overall, the combined study design provided a structured framework for the preclinical toxicological assessment of live biotherapeutic products.

Artificial intelligence (AI) is increasingly recognized as transformative in laboratory medicine, yet adoption lags behind radiology and cardiology. Limited data exist on AI-related attitudes and perceived barriers in Turkish clinical laboratories. This study evaluated professional awareness, attitudes, and perceived barriers to AI adoption among Turkish clinical laboratory professionals. A cross-sectional survey was conducted among Turkish clinical biochemistry staff between May and June 2025. The 30-item questionnaire covered demographics, AI knowledge, training, current use, future perspectives, and ethical or legal concerns. In total, 170 eligible participants, including laboratory directors, physicians, residents, and technologists, completed the survey. Among the participants, 43% self-rated their AI knowledge as "basic," with no participant aged ≥60 years reporting good or expert-level knowledge. The greatest knowledge deficits were in image processing (49.4% reported "no knowledge") and predictive analytics (35.9%). More than half (57.6%) had never received AI-related training, and current use of AI tools in routine practice was minimal. Despite these gaps, attitudes toward AI implementation were overwhelmingly positive, with 75.3% believing increased AI use would improve job satisfaction and 59.4% expecting laboratory transition to AI-supported systems within 5 years. Major barriers included insufficient digital infrastructure (47%), absence of trained personnel (83.5%), lack of standard operating procedures (70%), and inadequate legal/ethical regulations (52.9%). AI implementation in Turkish clinical laboratories remains at an early stage but is met with strong professional interest. Overcoming structural barriers through standardized education, infrastructure investment, and clear legal frameworks is essential for sustainable AI implementation in clinical laboratories.

Pathological conditions may result in clinical chemical values exceeding the linearity range of laboratory methods. In clinical practice, not all tests require exact numerical values for patient management, as useful information can still be provided when results are reported to a defined cutoff. This study describes an approach to reduce unnecessary dilutions by identifying tests that can be reported above a defined threshold. Eighty-six clinical chemistry assays on Cobas 8000 (middleware: Infinity) were reviewed at the Laboratory Medicine of the University Hospital of Padua. Fifteen analytes requiring manual dilution beyond predefined automated limits were selected. A multi-step workflow was implemented, including literature review, clinician and staff engagement, definition of upper reporting limits, middleware customisation, and retrospective and prospective performance evaluation. Key outcomes included turnaround time (TAT), auto-release rate, manual dilution frequency, estimated costs, and clinician requests for exact values. Literature review and clinician feedback supported the definition of upper reporting limits beyond which exact values were not routinely required, except for lactate dehydrogenase (LDH), where oncologic follow-up necessitated precise reporting. Implementation of the new rules eliminated most manual dilutions, substantially reducing costs and improving efficiency. P90 TAT decreased for samples above cutoff values in most analytes between 2023 and 2025. LDH was the only analyte for which manual dilution was not completely eliminated in 2025, with a residual rate of 3.38 per 1000 tests (vs. 7.32 per 1000 in 2023). Clinician feedback confirmed the appropriateness of the new reportingrules. This study shows that revising validation rules to report results above predefined thresholds provides clinically meaningful information while reducing unnecessary manual procedures. This model supports appropriate patient management and improves efficiency by enhancing process automation, consistency, and decision-making.

Monoclonal proteins (M-proteins) serve as critical biomarkers for plasma cell dyscrasias, yet their unique physicochemical properties paradoxically compromise the accuracy of routine laboratory diagnostics. Beyond causing well-documented anomalies in electrophoresis, M-protein interference pervasively affects coagulation, clinical chemistry, and immunoassay platforms, creating analytical artifacts that can fundamentally mislead clinical decision-making. The diagnostic landscape is further complicated by the widespread adoption of therapeutic monoclonal antibodies (t-mAbs), which introduce exogenous "drug spikes" mimicking disease markers and obscuring therapeutic response assessment. This narrative review elucidates the underlying mechanisms of these interferences, ranging from physical precipitation and chemical binding to immunological hook effects, and synthesizes evidence from recent clinical cohorts and technical evaluations. We critically assess the efficacy of mitigation strategies, highlighting the evolution from traditional techniques like polyethylene glycol precipitation to next-generation solutions involving mass spectrometry and artificial intelligence-driven algorithms. By establishing a comprehensive framework for the recognition and resolution of these artifacts, this article aims to equip clinicians and laboratorians with the tools necessary to ensure that laboratory diagnostics remain a pillar of patient safety.

Accurate molecular annotation is essential for deciphering biochemical processes in spatial biology. Here, we present a scalable and broadly applicable molecular annotation tool for tandem mass spectrometry imaging (MS2I). Our workflow includes parallel image acquisition (PIA) for parallel MS2I and an open-access computational framework for spatial similarity networking (SSN) that enables molecular annotation of MS2I data with isomeric specificity. The PIA enables simultaneous untargeted MSI and targeted MS2I ensuring structure-specific imaging of hundreds of molecules in a single experiment. The SSN increases annotation confidence through graph-based spatial correlation of product ion distributions, opening up new avenues for data investigation and annotation from both MSI and MS2I data. By integrating PIA and SSN into a single workflow, we visualize and annotate 134 phospholipid isomers and isobars in mouse brain tissue. Furthermore, we demonstrate the biological utility of the platform by mapping cholesterol metabolism in human multiple sclerosis brain tissue, revealing lesion-associated cholesterol oxidation pathways. Finally, we propose annotation confidence levels for structural annotation in MSI. Overall, PIA and SSN together provide large-scale, structure-specific MSI, expanding the scope for spatial metabolomics, lipidomics, and chemical pathology through molecular annotation beyond current capabilities.

The ability to cryopreserve and rewarm whole organs without damage has the potential to transform transplantation by enabling long-term banking and global organ sharing. Successful transplantation of rat kidneys cryopreserved and rewarmed using nanowarming, a technique in which silica coated iron oxide nanoparticles (sIONP) are perfused throughout the organ's vasculature along with cryoprotective agents (CPAs), which are then both washed out of the organ after rewarming prior to transplant has recently been reported. To support development towards clinical organ scale translation, the potential toxicity of sIONP in male Sprague-Dawley rats was evaluated, administering doses six to seven orders of magnitude higher than the estimated systemic exposure corresponding to the highest reported residual iron levels in organs following nanowarming and cryopreservation (3.36 ng Fe/kg). These doses are effectively within the range of a theoretical worst-case scenario, where a fully loaded kidney is transplanted (10.5-14mg Fe/kg) without washout. Toxicity was assessed at 24h for (5, 12, and 20mg Fe/kg) and over 28 days for 12mg Fe/kg dose. Furthermore, plasma pharmacokinetics, hematology, clinical chemistry, biodistribution and histopathology were evaluated. No overt toxicity was observed at the lower doses, while some adverse effects emerged at the highest dose of 20mg Fe/kg. These findings suggest minimal toxicity associated with sIONP injections of a dose equivalent to a fully loaded organ, supporting the translational potential of sIONP for organ nanowarming.

The present study evaluated the acute oral toxicity profile of microencapsulated semi-purified fibrinolytic proteases derived from Bacillus tequilensis HSFI-5 (Indonesian Patent Application No. S00202313485) using a modified OECD Guideline 423–aligned fixed-dose parallel design. BALB/c mice (n = 50; equal sex distribution) received single oral doses of 50, 300, 2000, or 5000 mg/kg body weight and were monitored for 14 days for clinical signs, body weight progression, hematological indices, clinical biochemistry parameters, organ weights, and gross pathological changes. No mortality or treatment-related clinical abnormalities were observed at any dose level. Body weight gain remained progressive and comparable to controls. Hematological and biochemical parameters showed no consistent dose-dependent deviations indicative of systemic inflammatory, metabolic, hepatic, or renal dysfunction. Absolute and relative organ weights, including liver and kidneys, remained within physiological ranges, and necropsy revealed no macroscopic lesions. Based on OECD decision criteria, the median lethal dose (LD50) was estimated to exceed 5000 mg/kg body weight, indicating that the formulation is not classified for acute oral toxicity under the Globally Harmonized System. Collectively, these findings indicate no evidence of acute treatment-related toxicological effects under the conditions of this study. Interpretation is limited to single-dose exposure and should be confirmed by further investigations incorporating detailed histopathological and extended toxicity assessments.

This article aims to define a minimum panel of essential laboratory tests to be used in emergency and disaster settings through expert consensus. A structured survey was distributed to the 24 members of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) Committee on Preparation of Laboratories for Emergencies (C-PLE). Participants were asked to rate a predefined list of diagnostic tests using a four-level priority scale (essential, critical, supportive, not recommended), with scores ranging from 1 to 4. Responses were collected over a two-week period, and mean scores with standard deviations were calculated to classify each test into priority categories. Additional test suggestions were also solicited and evaluated. A total of 20/24 members (83.3 %) completed the survey. Tests classified as essential (mean score 1.00-1.49) included complete blood count, electrolytes (sodium, potassium, chloride), blood glucose, lactate, blood gases (including ionized calcium), urinalysis, and ABO/RhD blood typing. Critical tests (mean score 1.50-2.49) comprised urea and/or creatinine, coagulation parameters such as prothrombin time/international normalized ratio (PT/INR) and activated partial thromboplastin time (APTT), aminotransferases (especially alanine aminotransferase; ALT), and cardiac troponins. Supportive tests (mean score 2.50-3.49) included C-reactive protein, D-dimer, rapid diagnostic tests for infectious diseases, and procalcitonin. Additional tests, such as urine pregnancy testing, bilirubin, and pancreatic enzymes, were proposed for potential inclusion based on clinical relevance. These findings establish a prioritized, consensus-based diagnostic framework tailored to resource-limited emergency contexts, emphasizing rapid, high-impact testing to support clinical decision-making and patient outcomes.

Accurate and precise pipetting is critical for reliable laboratory results, especially when handling small volumes or sensitive analytical techniques. Despite the common use of piston-operated pipettes, operator-dependent variability remains a major source of error. This study aimed to evaluate pipetting performance among laboratory personnel with varying levels of experience. The study included 108 participants: 26 laboratory school students (first to third year), 10 PhD students, 10 clinical chemists, and 62 laboratory technicians. Technicians were stratified into 3 groups based on the extent of sensitive manual pipetting in their work: minimal (Group A), moderate (Group B), and high (Group C). Pipetting performance was evaluated using gravimetric replicates at 10 µL and 100 µL volumes, and a dilution task involving serial dilutions of 1 M copper sulfate. Accuracy and precision metrics were compared across groups. The median coefficient of variation (CV%) was lower for 100 µL replicates (0.3% to 0.8%) than for 10 µL replicates (1.4% to 5.9%). Laboratory students showed the greatest variability and longest task completion times, although performance improved with advancing school year. Ten percent of participants exceeded ISO-defined accuracy limits for 10 µL pipetting, and 22% for 100 µL. Imprecision limits were surpassed by 36% and 20% of participants for 10 µL and 100 µL, respectively. Group C technicians achieved optimal dilution performance (R2 = 1.00), while students showed the highest deviation. Pipetting performance was significantly influenced by experience level and task sensitivity. Even experienced personnel occasionally failed to meet ISO standards, underscoring the need for ongoing training and performance assessment.

Game-based learning (GBL) has emerged as a promising approach to enhance student engagement and learning outcomes in various educational domains. This study aimed to assess the effect of applying GBL to biochemistry teaching on medical students' learning outcomes and performance, and to explore the students' perceptions and satisfaction. This study sought to determine whether embedding GBL in a second-year medical biochemistry module improves students' perceptions, engagement, and academic performance compared with traditional interactive teaching alone. A mixed research design was employed. A retrospective cohort design to assess and compare the results of achievement for the cohort of 2023/2024, which was instructed using GBL together with traditional interactive learning (exposed group), and the cohort of 2022/2023, which adopts traditional interactive pedagogy with the same learning outcomes, instructor, and question difficulty level) (the unexposed group) using the validated Measurement of Actual Knowledge and Engagement (MAKE) tool. Using GBL was positively received by most students. Simplification of biochemistry content (69.8%), led to students expressing interest in demonstrating their understanding (75.5%), engaging with biochemistry concepts (76.4%), and being stimulated to focus on the subject (83.5%). The results point toward the potential of GBL to improve learning outcomes with notable increases in skills and knowledge (70.2%), active participation (69.6%), and the application of concepts (65.5%). Integrating structured GBL activities into biochemistry teaching fostered higher student motivation and engagement and was associated with improved perceived learning and stable examination scores. Further controlled studies are warranted to elucidate causal links and long-term knowledge retention.

Circulating cell-free DNA (cfDNA) is valuable for molecular testing, but typically requires specialized collection tubes or immediate processing. We investigated whether residual plasma from heparin separators, routinely used in clinical chemistry, could serve as an accessible and underused source for cfDNA. We analyzed matched plasma samples from healthy volunteers in two experiments: an immediate-processing comparison across EDTA, Streck, and heparin separator tubes (n=5), and a clinical-handling simulation comparing EDTA and heparin separator tubes under delayed processing at room temperature or 4°C (n=6). We also analyzed matched plasma samples from viral PCR-positive patients in a hospital cohort (n=38). Whole-genome sequencing and enriched methylation sequencing were performed to assess concordance across metagenomics, copy number, methylation, and fragmentomic features. Under immediate processing, heparin separator plasma showed high concordance with EDTA and Streck plasma for methylation patterns (Spearman's ρ=0.65-0.70) and fragmentation features. In the Hospital Cohort, heparin separator plasma showed strong concordance with matched EDTA plasma for viral detection (Spearman's ρ=0.95), copy number alteration profiling (Spearman's ρ=0.72-0.96), and methylation patterns (Spearman's ρ=0.50-0.83). These findings support the feasibility of using refrigerated, promptly processed residual plasma from routine clinical chemistry as a supplementary source for cfDNA biobanking and molecular analyses.

Introduction: Serum electrolyte measurement, particularly sodium, potassium, and chloride, is among the most commonly ordered investigations in routine and emergency care. Rapid and accurate reporting is essential for timely clinical decisions. Most modern electrolyte analysers utilise Ion-selective Electrode (ISE) technology, available in two forms: direct ISE (dISE), which measures electrolytes in undiluted serum, and indirect ISE (iISE), which measures them after pre-dilution. These methodological differences may influence reported values. Aim: To compare the serum sodium, potassium, and chloride levels measured using dISE versus iISE methods. Materials and Methods: The cross-sectional study was conducted in the Clinical Biochemistry Laboratory, Government Medical College and Hospital, Chandigarh, India, from March 2024 to and June 2024. A total of 150 serum samples were obtained from the outpatients, inpatients, and Intensive Care Unit (ICU) patients. Paired serum samples available for simultaneous direct and iISE estimation were included. Deidentified residual samples were analysed and assigned a unique code. dISE was performed using the ABL800 FLEX (Radiometer) on undiluted serum, while iISE was performed using the Beckman Coulter AU5800 on prediluted serum; both followed the manufacturer’s calibration and two-level internal Quality Control (QC) protocols. Testing order was randomised, and paired measurements were completed within 30-60 minutes to avoid analyte instability. Statistical analysis included Bland–Altman plots to assess agreement and Spearman’s correlation to evaluate the association between the two methods. Analysis was stratified by clinically relevant electrolyte ranges, with p<0.05 considered statistically significant. Results: dISE yielded significantly higher sodium and chloride levels than iISE. Subgroup analysis using clinically relevant thresholds demonstrated significant correlations for serum sodium ≤140 mEq/L, potassium ≥4.5 mEq/L, and chloride levels across both concentration groups (>103 and <103 mEq/L), with correlation coefficients of r=0.546, 0.870, 0.602, and 0.703, respectively (p<0.0001). Conclusion: A significant difference was observed between dISE and iISE methods for serum electrolyte measurement, particularly within specific clinical ranges. These findings highlight the need for methodological consistency in electrolyte analysis, especially in emergency and critical care settings, to prevent misinterpretation and inappropriate clinical decisions arising from analytical variability.