Clinical Chemistry Analyzer Research Articles

First report of Cytauxzoon sp. infection in Germany: organism description and molecular confirmation in a domestic cat.

Cytauxzoonosis is described as an emerging tick-borne disease of domestic and wild felids caused by protozoans of the genus Cytauxzoon. While in the Americas the condition is described as a fatal disease, in Europe, reports on the clinical expression of the infection are scarce. This study describes the first case of Cytauxzoon sp. infection in Germany, in a domestic cat. A 6-year-old male domestic cat living in Saarlouis (Saarland) was presented with anorexia, lethargy and weight loss. The cat had an outdoor lifestyle and had not travelled abroad. Serum clinical chemistry analysis revealed azotaemia with markedly increased symmetric dimethylarginine, hypercreatinemia, hyperphosphatemia and hypoalbuminemia. Moreover, a mild non-regenerative anaemia was present. Approximately 1 year prior to these findings, the domestic cat was diagnosed with a feline immunodeficiency virus (FIV) infection. These results pointed toward a decreased glomerular filtration rate, presumably as a result of kidney dysfunction. Round to oval signet ring–shaped intraerythrocytic organisms, morphologically suggestive for a piroplasm, were revealed during blood smear evaluation with a degree of parasitaemia of 33.0%. PCR analyses and sequencing of a region of the 18S rRNA gene confirmed the presence of a Cytauxzoon sp. infection, with 99–100% nucleotide sequence identity with previously published Cytauxzoon sp. isolates. As this is the first molecularly confirmed Cytauxzoon sp. infection in a domestic cat in Germany, these findings suggest that cytauxzoonosis should be considered as a differential diagnosis in cases of anaemia in outdoor domestic cats, particularly in areas where wild felid populations are present.

Obesity and cardiovascular risk among Sri Lankan adolescents: Association of adipokines with anthropometric indices of obesity and lipid profile.

Obesity and being overweight among adolescents pose a significant problem and are known to cause several physical and biochemical disorders during adulthood. This study was designed to identify the biomarkers of obesity and describe associations with selected metabolic disorders of obesity among Sri Lankan adolescents. The present study compared the characteristics of obese (n=121) and normal weight (n=263) adolescents, including sociodemographic, anthropometric, and selected biochemical parameters (e.g., lipid profile, serum leptin, adiponectin, and high-sensitivity C-reactive protein [hs-CRP]). An enzyme-linked immunosorbent assay technique and fully automated clinical chemistry analyzer were used to analyze the biochemical parameters among adolescents ages 10 to 16. The mean age of the sample was 13.1 y [standard deviation (SD): 1.9 y], and the male-to-female ratio 1:1. The mean weight of obese children was 55.70 kg (SD: 14.82 kg), which was significantly higher than that of children of normal weight [41.63 kg (SD: 7.88 kg)]. Total cholesterol, triacylglycerol, and low-density lipoprotein cholesterol levels were significantly higher (P=0.000) among obese adolescents compared with those of normal weight. High-density lipoprotein cholesterol was significantly lower among obese adolescents. Serum leptin and hs-CRP were higher among obese adolescents, but adiponectin was lower. In the multivariate analysis, owing to confounding effects among the tested adipokines, serum leptin was the only predictor of an abnormal lipid profile. Serum leptin, adiponectin, and hs-CRP were found to be reliable biomarkers of predicting adiposity related metabolic disorders in adolescents. Obese adolescents showed disorders in the lipid metabolism with abnormal lipid profiles compared with children of normal weight.

Analysis of serum glutamic pyruvic transaminase and serum glutamic oxaloacetic transaminase levels in tuberculosis patients who are undergoing oat treatment in Kendari City General Hospital, Kota Kendari, Indonesia.

In the treatment of tuberculosis (TB), the patient is generally directly given Anti Tuberculosis Drugs (Obat Anti Tuberculosis/OAT) without examining Serum Glutamic Pyruvic Transaminase (SGPT) and Serum Glutamic Oxaloacetic Transaminase (SGOT) to see whether or not there is liver damage before treatment. Because of the side effects by OAT, it is important to know how the condition of the liver function of TB patients who consume OAT in Kendari City General Hospital (Rumah Sakit Umum Daerah/RSUD) Kota Kendari by looking at SGOT and SGPT levels in order to provide maximum treatment to TB patients. The method of this research was descriptive analysis with cross sectional approach. The sample is all patients had previously been diagnosed with TB by a doctor based on medical records by in RSUD Kota Kendari after 1-2 months OAT treatment without any sign of liver injury before. Examination of SGOT and SGPT levels was performed on all samples in the Laboratory of RSUD Kota Kendari using clinical chemistry analyzer. The results of this study showed that 20% patients with TB had elevated levels of SGPT and SGOT with average SGOT is 51 U/L and SGPT is 42.5. The Inference of this study that there is an increase in SGOT and SGPT levels in patients who consume OAT 1-2 months in RSUD Kota Kendari. Elevated levels of SGPT and SGOT due to Anti Tuberculosis (OAT) drugs did not occur in all patients and only increased in 3 people, but qualitatively the effect of administering anti-tuberculosis drugs significantly increased levels of SGPT and SGPT. Thus, monitoring of liver physiology remains recommended especially for elderly patients.

Acute and Sub-chronic Toxicity of Indole Alkaloids Extract from Leaves of Alstonia scholaris (L.) R. Br. in Beagle Dogs

Alstonia scholaris (L.) R. Br., an evergreen tropical plant rich in indole alkaloids with significant physiological activity, is traditionally used to treat respiratory diseases in China. This study was conducted to establish the toxicity profile of the alkaloid extract (TA) of A. scholaris leaves in non-rodents. After oral administration of a single dose (4 g/kg.bw), a number of transient symptoms, such as unsteady gait, drooling, emesis, and reddening of peri-oral mucosa, were observed, but no treatment-related mortality. A sub-chronic toxicity study with a range of doses of TA (20, 60 and 120 mg/kg.bw) was conducted for a 13-week treatment period, followed by 4-week recovery observation. Except for emesis and drooling in majority of animals in 120 mg/kg.bw treatment group, no clinical changes were observed in TA-treated animals. Data from electrocardiography, bone marrow, urine, fecal, hematology and clinical chemistry analyses were comparable between TA-treated and control animals. No significant differences in the relative organ weights and histopathological characteristics were evident between the TA-treated and control groups. Accordingly, the non-observed-adverse-effect-level (NOAEL) of TA was established as 120 mg/kg.bw. Our results add further knowledge to the safety database for indole alkaloid extracts from A. scholaris with potential utility as novel drug candidates.Graphic

Prevalence and Determinants of Hyperuricemia in Type 2 Diabetes Mellitus Patients Attending Jimma Medical Center, Southwestern Ethiopia, 2019.

ObjectiveTo assess the prevalence and determinants of hyperuricemia among type 2 diabetic patients on follow-up at Jimma Medical Center from March 1 to June 30, 2019.MethodsThis institution-based cross-sectional study was used to assess the level of serum uric acid in type 2 diabetic patients attending their follow-up at Jimma Medical Center chronic illness clinic. A systematic sampling technique was used to include 287 type 2 diabetic patients in the study and an interviewer-based questionnaire was used to collect important data from each participant. Five milliliters of blood were collected from each participant under sterile conditions and serum was separated by centrifugation at 3000 rpm for 10 minutes. Serum was stored at −20°C and analyzed for serum uric acid using an ABX Pentra 400 clinical chemistry analyzer. Bivariate and multivariable binary logistic regression were used to assess significant associations between hyperuricemia and independent factors. A p-value of <0.05 was considered significant in the final model.ResultsThe mean±SD age of the study participants was 51.79±14.36 years. The prevalence of hyperuricemia was found to be 22% (n=66/287) in the study population. Hyperuricemia was common in those aged ≥60 years and males. Obesity (AOR=7.84, 95% CI=2.005–30.666), duration of diabetes mellitus (DM) ≥10 years (AOR=3.963, 95% CI=1.902–8.259), family history of CVD (AOR=2.929, 95% CI=1.124–7.630), alcohol drinking (AOR=5.83, 95% CI=2.341–14.545) and increased DBP (AOR=4.198, 95% CI=1.772–9.949) were determinant variables for hyperuricemia in type 2 DM.ConclusionHyperuricemia was relatively common among type 2 diabetic patients. The prevalence of hyperuricemia was common among patients with obesity, a long duration of DM and increased diastolic blood pressure, and alcohol drinkers. There is a need to raise awareness of lifestyle modification, healthy behavior and early diagnosis of hyperuricemia in type 2 diabetic patients.

Chemiluminescence Biosensor for Non-invasive Crew Health Monitoring at the International Space Station

Long-duration manned space missions require that astronauts be equipped with analytical devices suitable for performing on-site clinical-chemistry analysis. In the frame of the ???IN SITU Bioanalysis??? project, an analytical device for measuring the steroid hormone cortisol (an indicator of chronic stress) in saliva has been developed. The device performed a Lateral Flow Immunoassay (LFIA) for cortisol employing disposable plastic cartridges produced by FDM (Fused Deposition Modeling) 3D printing technology, which encapsulated a sealed fluidic element containing the LFIA strip and the reagents required for the analysis. Detection was performed by chemiluminescence (CL) using a dedicated CL reader equipped with a cooled charge-coupled device (CCD) camera. The European Space Agency (ESA) astronaut Paolo Nespoli successfully used the ???IN SITU Bioanalysis??? analytical device during the mission ???VITA???, a 6-months space mission onboard the International Space Station (ISS) in the framework of a dated bilateral agreement between the Italian Space Agency (ASI???Agenzia Spaziale Italiana) and NASA. The experiments proved the feasibility of CL-LFIA ultrasensitive (i.e., at the sub-ng/mL level) quantitative bioassays in space. In perspective, the ???IN SITU Bioanalysis??? analytical device could be adapted to the measurement of other biomarkers of diagnostic and prognostic relevance, even in a multiplexed fashion (i.e., detection of multiple biomarkers in a single analysis) to develop panels tests for diagnosis and follow-up of pathologies or monitoring health status of astronauts.

SAT-396 A Non-Surgical Animal Model of Hypoparathyroidism for Testing PTH Analogues

BACKGROUND:In vivo animal models for testing the pharmacokinetics and bioactivity of PTH and its analogues require parathyroidectomy by surgery (1, 2). As the parathyroid glands of rodents are very small the surgery often includes thyroidectomy, making this animal model time-limited, single use, complex, and expensive. We have developed a non-surgical rodent model of hypoparathyroidism using the Type II calcimimetic compound, Cinacalcet-HCl, to suppress PTH and thereby serum calcium levels.Methods:Normal male Wistar rats were gavaged with 30 mg/kg Cinacalcet-HCl (or vehicle only). To test the effect of PTH 1–34, animals were dosed immediately after Cinacalcet-HCl gavage with either a single subcutaneous injection of PTH at 20 nmol/kg or given as same dose repeated every hour for 6 hrs or vehicle only. Serum samples were analysed for ionised calcium (iCa) using an EasyLyte, fully automated electrolyte analyser (Medica Corporation) and phosphate using a Phosphorus Detection Assay Kit (Pars Azmun, IRAN) and an Hitachi 917 Clinical Chemistry Analyser.Results:Rats gavaged with 30 mg/kg Cinacalcet-HCl produced a significant reduction in iCa levels between 2-24hrs returning to baseline at 48-72hrs post dose with the nadir at 8 hours (ANOVA P < 0.0001). This equated to a 25% reduction in iCa at 8 hrs: mean±SD, iCa 1.19 ± 0.09 mmol/L at predose and 0.891 ± 0.04 mmol/L at 8 hours (t-test P < 0.0001). For phosphate there was an initial lowering within the first 2 hrs in all test groups but then a rise such that phosphate was at higher levels than control from 8–24 hrs (ANOVA, ns), returning to baseline at 48 hrs. PTH at 20 nmol/kg given as a single sc dose abrogated the Cinacalcet-HCl induced fall in iCa for up to 2 hrs (AUC±SD (mmol/L).hr, 0.076 ±0.047 versus 0.168±0.0874, t-test P=0.0289).Conclusions:We have shown that the administration of Cinacalcet-HCl provides a robust and reproducible lowering of calcium which is line with current published data (3). These studies demonstrate that the use of Cinacalcet-HCl in normal rats produces a hypocalcemic state that can be abrogated by the addition of PTH. This non-surgical animal model of hypoparathyroidism will be of value in testing the pharmacodynamics of PTH analogues.

Evaluation of the Liver Enzyme (AST, ALT &amp; ALP) Levels of Adult HIV Patients on HAART in UPTH

This study evaluated the liver enzyme levels of HIV-infected adult patients on highly active antiretroviral treatment (HAART) for not less than 1 year. The study was designed as a randomised cross-sectional study that evaluated the differences in the aspartate aminotransferase [AST], alanine transaminase [ALT] and alkaline phosphatase [ALP] of 129 (46 males and 83 females) HIV-infected adult patients. Before the study, ethical clearance (Ref: UPH/CEREMAD/REC/19) was obtained from the University of Port Harcourt Ethics Committee. Venous blood samples were obtained and the liver enzyme levels were analysed using Clinical Chemistry Analyser (VS10) manufactured by Vitro Scient. Values were further categorised into; normal or abnormal. SPSS version 21 (IBM® Armonk, USA) and Graph Pad Prism (Version 8.0.2) was used to analyse the data. T-test compared the sex differences in mean, while Chi-square analysis tested the sex differences in the categorised data. From the result, the mean AST (27.65±17.93 IU/L) and ALP (135.13±10.87 IU/L) values for males were higher than females AST (27.25±18.93 IU/L) and ALP (132.65±9.96 IU/L) values, while ALT was higher in female (34.66±22.29 IU/L) than males (33.75±18.14 IU/L); however, the differences were not significant (p&gt;0.05). Generally, abnormal AST, ALT and ALP levels were 31%, 34% and 82% respectively; with no sex-associated differences (p&gt;0.05). 45.7% of the patients (males: 25 [54.3%] and females: 34 [41.0%]) had cholestatic abnormality, while 8.5% (5; 10.9% males and 6; 7.2% females) had hepatocellular abnormality, 18.6% (6; 13.0% of males and 18; 21.7% of females) and mixed abnormality (AST/ALT/ALP). In conclusion, cholestatic abnormalities were observed in more than 80% of the patients as opposed to hepatocellular abnormalities, which were less than 35%. More females were associated with mixed abnormality when compared to males with independent (ALT/AST) abnormality. This study, therefore, suggests the need for a randomised case-control study to highlight the extent of deviation from normal values.

Cover, Content, and Editorial Note from Borneo J Med Lab Tech Vol. 2 No. 2 April 2020

Assalamu�alaikum Wr. Wb.Alhamdulillahirabbil �alamin. After a long wait for almost 12 years from the first stand, finally the scientific journal of the Department of Analyst Health (Medical Laboratory Technology) Universitas Muhammadiyah Palangkaraya can be published. BJMLT published every 6 months (2 issues/year) every October and April. The aim of this journal is to publish high-quality articles dedicated to all aspects of the latest outstanding developments in the field but not limited to Blood transfusion, Clinical Chemistry, Hematology, Histopathology, Immunology, Microbiology, Parasitology, Toxicology, Food and Beverage Analysis, Molecular Biology, Medical Biology, and related fields that are of interest to medical laboratory technologies, such as epidemiology, health promotion, professional ethics, and laboratory management, It was first published online in October 2018 usingOpen Journal System (OJS) 3.1 platform with e-ISSN 2622-6111 by url http://journal.umpalangkaraya.ac.id/index.php/bjmlt.This edition contains 6 articles consisting of Hematology, Microbiology, Clinical Chemistry and epidemiology topics. Editorial boards are fully aware that there are still room for improvement in this edition, hence with all humility willing to accept constructive suggestions and feedback for improvements to the publication for the next editions.The publication of BJMLTcertainly participate in disseminating the results of research and review of science and technology development conducted by lecturers and researchers from UM Palangkaraya and other universities.The editorial board would like to thank the University, all editors and reviewers, and contributors of the scientific articles who have provided the repetoire in this issue. We hope that all parties, especially the contributors of the articles, could re-participate for the success of the publication in the next edition on october 2020.Wassalamu�alaikum Wr. Wb.Palangka Raya, April 2020Editor-in-Chief

Community based reference interval of selected clinical chemistry parameters among apparently healthy Adolescents in Mekelle City, Tigrai, Northern Ethiopia.

Locally established clinical laboratory reference intervals (RIs) are required to interpret laboratory test results for screening, diagnosis and prognosis. The objective of this study was establishing reference interval of clinical chemistry parameters among apparently healthy adolescents aged between 12 and 17 years in Mekelle, Tigrai, northern part of Ethiopia. Community based cross sectional study was employed from December 2018 to March 2019 in Mekelle city among 172 males and 172 females based on Multi stage sampling technique. Blood samples were tested for Fasting blood sugar (FBS), alanine aminino transferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP), Creatinine, urea, total protein, albumin (ALB), direct and indirect bilirubin (BIL.D and BIL.T) using 25 Bio system clinical chemistry analyzer. Results were analyzed using SPSS version 23 software and based on the Clinical Laboratory Standard Institute (CLSI)/ International Federation of Clinical Chemistry (IFCC) C 28-A3 Guideline which defines the reference interval as the 95% central range of 2.5th and 97.5th percentiles. Mann Whitney U test, descriptive statistics and box and whisker were statistical tools used for analysis. This study observed statistically significant differences between males and females in ALP, ALT, AST, Urea and Creatinine Reference intervals. The established reference intervals for males and females, respectively, were: ALP (U/L) 79.48-492.12 versus 63.56-253.34, ALT (U/L) 4.54-23.69 versus 5.1-20.03, AST 15.7-39.1 versus 13.3-28.5, Urea (mg/dL) 9.33-24.99 versus 7.43-23.11, and Creatinine (mg/dL) 0.393-0.957 versus 0.301-0.846. The combined RIs for Total Protein (g/dL) was 6.08-7.85, ALB (g/dL) 4.42-5.46, FBS(mg/dL) 65-110, BIL.D (mg/dL) 0.033-0.532, and BIL.T (mg/dL) 0.106-0.812. The result showed marked difference among sex and with the company derived values for selected clinical chemistry parameters. Thus, use of age and sex specific locally established reference intervals for clinical chemistry parameters is recommended.

The effects of shock wave lithotripsy and retrograde intrarenal surgery on renal function

Introduction The aim of this study was to compare the early effects of shock wave lithotripsy (SWL) and retrograde intrarenal surgery (RIRS) on renal function using the cystatin C levels. Material and methods Serum samples were taken from each of the patients preoperatively, on the first postoperative day, and on the 30th postoperative day in order to evaluate the renal damage. The cystatin C level was determined using a particle-enhanced turbid metric immunoassay with a clinical chemistry analyzer. Results In the comparison between the preoperative and postoperative cystatin C levels on day 1, there was an increase in the SWL group (p = .001); however, the decrease in the RIRS group was statistically significant (p = .007). There were statistically significant differences in the cystatin C levels on the first postoperative day in both groups (p = .001). In the SWL group, there was a statistically significant increase between the preoperative and the 30th postoperative day cystatin C levels (p = .006), but no differences were found between these levels in the RIRS group or between the two groups (p = .255). Conclusions RIRS may be the preferred procedure for patients who need more renal function protection when treating renal stones <2 cm

Gender-Dependent Association of Vitamin D Deficiency with Obesity and Hypercholesterolemia (LDLC) in Adults.

Obesity, dyslipidemia and vitamin D deficiency are growing health problems in the Arabian Gulf region. Their association with each other is yet to be clarified. Three-hundred and fourteen Bahraini adults, 164 males and 150 females comparable in median age (34.5 vs. 31.0 yrs), body mass index (BMI), and ethnicity were recruited. The plasma level of 25-hydroxyvitamin D3 (25OHD3) was measured by chemiluminescent immunoassay and lipid profile parameters were measured by an automated clinical chemistry analyzer. Based on BMI, study subjects were grouped into underweight, normal, overweight, moderate obesity, and severe obesity subjects. The results revealed an extremely high prevalence of vitamin D deficiency (79.9%) and insufficiency (18.8%). The predictors of low 25OHD3 levels were female gender, small age, conservative dressing, least exposure to sunlight, and less fish intake. In all subjects, the lowest 25OHD3 level was seen in underweight and severe obesity groups. Furthermore, the 25OHD3 level was significantly higher in males as compared to females and it was positively correlated with the age. However, detailed analysis showed that overweight males unlike females had the highest 25OHD3 levels which were significantly higher than in the severely obese males. While the lipid profile parameters were positively correlated with BMI, the total and LDL cholesterols were negatively correlated with the levels of 25OHD3 in males. Vitamin D deficiency was associated with both severely obese and underweight subjects, in the former it was likely to be institutional while in the latter it was likely to be nutritional. Furthermore, hypercholesterolemia (LDL-C) was associated with 25OHD3 sub-normality. Further analysis revealed that the significant associations were gender-dependent.

SEX DIFFERENCES IN THE REFERENCE INTERVALS OF HEALTH PARAMETERS IN SEMICAPTIVE ASIAN ELEPHANTS (ELEPHAS MAXIMUS) FROM MYANMAR.

The reference intervals of health parameters are valuable tools for veterinarians and conservationists to monitor the health status and viability of endangered species. Natural variation in the health of the long-lived Asian elephant (Elephas maximus) is poorly understood, particularly in relation to differences between males and females. Longitudinal health data were collected from clinical examination, hematology, and serum chemistry analyses over 3 yr from 227 healthy individually marked Asian elephants varying in age and sex. The study population was semicaptive and used in Myanmar's timber industry, but maintained natural feeding and breeding behavior. Body condition score (BCS) and blood pressure were investigated in clinical examinations. Hematological parameters included hematocrit, hemoglobin, total white blood cell count, and differential blood cell counts. Serum chemistry parameters included blood urea nitrogen, creatinine, total protein, albumin, globulins, aspartate aminotransferase, alkaline phosphatase, triglycerides, creatine kinase, glucose, calcium, potassium, sodium, and chloride. To the knowledge of the authors, this is the first description of BCS in an elephant population outside of zoos, and of blood pressure in this species using a novel adaptation of the Intelli Wrap Cuff pressure monitor. Several differences between the sexes were observed, with females generally having higher BCS and triglycerides, and males displaying higher alkaline phosphatase and glucose levels. This study provides important clinical tools that can be used to assess the health status and improve management in this endangered species.

A Dual Workflow to Improve the Proteomic Coverage in Plasma Using Data-Independent Acquisition-MS.

Plasma is one of the most important and common matrices for clinical chemistry and proteomic analyses. Data-independent acquisition (DIA) mass spectrometry has enabled the simultaneous quantitative analysis of hundreds of proteins in plasma samples in support population and disease studies. Depletion of the highest abundant proteins is a common tool to increase plasma proteome coverage, but this strategy can result in the nonspecific depletion of protein subsets with which proteins targeted for depletion interact, adversely affecting their analysis. Our work using an antibody-based depletion column revealed significant complementarity not only in the identification of the proteins derived from depleted and undepleted plasma, but importantly also in the extent to which different proteins can be reproducibly quantified in each fraction. We systematically defined four major quantitative parameters of increasing stringency in both the depleted plasma fraction and in undepleted plasma for 757 observed plasma proteins: Linearity cutoff r2 > 0.8; lower limit of quantification (LLOQ); measurement range; limit of detection (LOD). We applied the results of our study to build a web-based tool, PlasmaPilot, that can serve as a protocol decision tree to determine whether the analysis of a specific protein warrants IgY14 mediated depletion.

Understanding acute metabolic decompensation in propionic and methylmalonic acidemias: a deep metabolic phenotyping approach

BackgroundPathophysiology of life-threatening acute metabolic decompensations (AMD) in propionic acidemia (PA) and isolated methylmalonic acidemia (MMA) is insufficiently understood. Here, we study the metabolomes of PA and MMA patients over time, to improve insight in which biochemical processes are at play during AMD.MethodsLongitudinal data from clinical chemistry analyses and metabolic assays over the life-course of 11 PA and 13 MMA patients were studied retrospectively. Direct-infusion high-resolution mass spectrometry was performed on 234 and 154 remnant dried blood spot and plasma samples of PA and MMA patients, respectively. In addition, a systematic literature search was performed on reported biomarkers. All results were integrated in an assessment of biochemical processes at play during AMD.ResultsWe confirmed many of the metabolite alterations reported in literature, including increases of plasma valine and isoleucine during AMD in PA patients. We revealed that plasma leucine and phenylalanine, and urinary pyruvic acid were increased during AMD in PA patients. 3-hydroxyisovaleric acid correlated positively with plasma ammonia. We found that known diagnostic biomarkers were not significantly further increased, while intermediates of the branched-chain amino acid (BCAA) degradation pathway were significantly increased during AMD.ConclusionsWe revealed that during AMD in PA and MMA, BCAA and BCAA intermediates accumulate, while known diagnostic biomarkers remain essentially unaltered. This implies that these acidic BCAA intermediates are responsible for metabolic acidosis. Based on this, we suggest to measure plasma 3-hydroxyisovaleric acid and urinary ketones or 3-hydroxybutyric acid for the biochemical follow-up of a patient’s metabolic stability.

Pre-/analytical factors affecting whole blood and plasma glucose concentrations in loggerhead sea turtles (Caretta caretta).

Blood glucose is vital for many physiological pathways and can be quantified by clinical chemistry analyzers and in-house point-of-care (POC) devices. Pre-analytical and analytical factors can influence blood glucose measurements. This project aimed to investigate pre-analytical factors on whole blood and plasma glucose measurements in loggerhead sea turtles (Caretta caretta) by evaluating the effects of storage (refrigeration) up to 48h after sampling and of packed cell volume (PCV) on whole blood glucose analysis by POC glucometer (time series n = 13); and by evaluating the effects of storage (room temperature and refrigeration) on plasma glucose concentrations using a dry slide chemistry analyzer (DCA) at various conditions: immediate processing and delayed plasma separation from erythrocytes at 24h and 48h (time series n = 14). The POC glucometer had overall strong agreement with the DCA (CCC = 0.76, r = 0.84, Cb = 0.90), but consistently overestimated glucose concentrations (mean difference: +0.4 mmol/L). The POC glucometer results decreased significantly over time, resulting in a substantial decline within the first 2h (0.41±0.47 mmol/L; 8±9%) that could potentially alter clinical decisions, thereby highlighting the need for immediate analysis using this method. The effects of PCV on glucose could not be assessed, as the statistical significance was associated with one outlier. Storage method significantly affected plasma glucose measurements using DCA, with room temperature samples resulting in rapid decreases of 3.57±0.89 mmol/L (77±9%) over the first 48h, while refrigerated samples provided consistent plasma glucose results over the same time period (decrease of 0.26±0.23 mmol/L; 6±5%). The results from this study provide new insights into optimal blood sample handling and processing for glucose analysis in sea turtles, show the suitability of the POC glucometer as a rapid diagnostic test, and confirm the reliability of plasma glucose measurements using refrigeration. These findings emphasize the need to consider pre-/analytical factors when interpreting blood glucose results from loggerhead sea turtles.

Correlations between Direct and Calculated Low-Density Lipoprotein Cholesterol Measurements in Children and Adolescents.

BackgroundLow‐density lipoprotein cholesterol (LDL‐C), as a modifiable risk factor for atherosclerotic cardiovascular disease, should be assessed and monitored. This study compared directly measured and Friedewald‐estimated LDL‐C values in children and adolescents.MethodsBlood samples were collected from 464 children and adolescents. Calculated LDL‐C (CLDL‐C) levels were estimated using the Friedewald formula for any triglyceride value below 4.6 mmol/L. Direct LDL‐C (DLDL‐C) levels were measured on an ARCHITECT c8000 Abbott Clinical Chemistry Analyzer. The differences in LDL‐C were then calculated.ResultsThe correlation coefficients (R) between DLDL‐C and CLDL‐C were 0.978 (P = .148) and R = 0.970 (P = .052) for children and adolescents, respectively. Children with LDL‐C values above 4.92 mmol/L had a correlation value of 0.971 (P = .419). The correlation and agreement between DLDL‐C and CLDL‐C in adolescents were moderate for LDL‐C below 2.85 mmol/L (R = 0.806; 84.1%) and improved above 2.85 mmol/L (R = 0.978; 91.5%). In children, good correlations between DLDL‐C and CLDL‐C were observed for normal (<0.85 mmol/L), borderline (0.85‐1.12 mmol/L), and abnormal (≥1.13 mmol/L) triglyceride levels (R = 0.9782, 0.990, and 0.951, respectively). The rates of agreement were better for normal (80.5%) and borderline (82.9%) but not abnormal (68.2%) triglyceride levels.ConclusionWe observed good agreement between DLDL‐C and CLDL‐C in both children and adolescents. The Friedewald formula provided an adequate estimate of LDL‐C for most fasting specimens. LDL‐C difference percentage can also be used as a quality indicator to check laboratory analyzer performance in healthy subjects.

Review of interference indices in body fluid specimens submitted for clinical chemistry analyses

ObjectivesThe aims of this study were to retrospectively investigate interference indices in a wide range of body fluid specimens and compare these indices to those found in serum/plasma. Design and MethodsThis retrospective study evaluated interference indices for hemolysis, icterus, and lipemia in 2752 body fluid specimens submitted for clinical chemistry testing. ResultsThe distribution of interference indices for body fluid samples was generally similar to that of serum/plasma interference indices. Hemolysis of specimens submitted for lactate dehydrogenase (LD) represented the most common interference for body fluid chemistries. Body fluids collected from postsurgical drain sites had a higher proportion of tests exceeding both icterus and lipemic limits compared to serum/plasma specimens. ConclusionsOverall, degrees of hemolysis, icterus, and lipemia observed in body fluid specimens were in large part similar to serum/plasma specimens, with a few notable differences. Body fluids exhibited a higher proportion of samples with severe icterus or lipemia. Severely lipemic body fluid samples were significantly less likely to also be hemolyzed relative to severely lipemic serum/plasma specimens. LD was the test most commonly affected by interference across all body fluid types. False elevations in pleural fluid LD induced by hemolysis can lead to mis-classification of transudative effusions as exudative using Light’s criteria. The possible impact of interferences on clinical chemistry testing in body fluids is an important post-analytical consideration.

Validation of a novel and accurate ApoE4 assay for automated chemistry analyzers

The allele ε4 of the apolipoprotein E gene (APOE ε4) is the major genetic risk factor for non-dominantly inherited Alzheimer’s Disease (AD). Current techniques for APOE ε4 carriers identification show good accuracy but have several disadvantages that limit its implementation in a clinical laboratory. These include the need for sample preprocessing, poor automation, low throughput, requirement of additional equipment, and high cost. We followed ISO 13485 guidelines to validate the e4Risk test, a new latex-enhanced immunoturbidimetric blood assay for apolipoprotein E4 (ApoE4) determination in human plasma samples. The test showed high performance in terms of lot to lot variability, precision, interferences, reagents stability, prozone, and detectability. Furthermore, diagnostic accuracy is almost equal (99%) to the gold standard, APOE ε4 genotyping by polymerase chain reaction (PCR). Furthermore, we demonstrated that the e4Risk test can be adapted to any clinical chemistry analyzer, including the high throughput analyzers present in most hospitals and clinical laboratories. The e4Risk test versatility, low cost, and easiness provides an excellent solution for APOE ε4 carriers identification using the same blood sample drawn for biochemical diagnostic work-up of AD patients, which can have important advantages for patient stratification in clinical trials, preventative strategies for AD, and clinical assessment of risk for brain amyloidosis.

A sample-to-answer quantitative platform for point-of-care testing of biochemical markers in whole blood

Point-of-care quantitative detection of biochemical markers has broad applications in resource-limited settings. However, current challenges including difficulty in sample preparation and limitation in result quantitation make point-of-care detection platforms failed to achieve fully sample-to-answer detection or meet the clinical quantitative demands. Here, we developed a novel point-of-care platform to realize fully sample-to-answer biochemical marker quantitation in whole blood. Our platform integrated a plasma separation module and a detection module. The plasma separation module contained a plasma separation strip could rapidly separate the plasma from whole blood samples by lateral flow. The detection module generated a colorimetric signal which could be quantified by the smartphone ambient light sensor (ALS) in an equipment-free manner. As a proof-of-concept, two hepatobiliary disease-associated markers, total bilirubin (TBil) and direct bilirubin (DBil), were detected from whole blood to quantitative result. The quantitative performance for clinical samples was highly consistent with the well-established clinical chemistry analyzer. Our platform exhibited remarkable advantages of significant portability (< 40 g), low cost (< $5), rapidity (< 5 min), instrument-free, high sensitivity (< 1 μM), and accuracy (89.5 % for TBil, 94.7 % for DBil), which showed up a great potential for biochemical markers detection in resource-limited settings.