Total CV Research Articles

Inter-laboratory variation for urine albumin among laboratories in a Swedish external quality assessment scheme 2005–2023

IntroductionIncreased albuminuria is associated with elevated mortality. Urine albumin (U-ALB) above 20 mg/L or albumin-to-creatinine ratio (U-ACR) of 3 g/mol are indicative of moderately increased albuminuria. Due to limited standardization among U-ALB methods, diagnosis of increased albuminuria might prove difficult. Materials and methodsData from Equalis’s external quality assessment scheme for low U-ALB levels during 2005–2023 were categorized according to manufacturer and divided into central laboratory (CLAB) and point-of-care testing (POCT) methods. Manufacturer median values were compared to total group mean consensus values and manufacturer CV% was compared at different U-ALB levels. ResultsCLAB was generally closer to consensus values and had lower CV% than POCT at U-ALB levels around 20 mg/L. For CLAB, Roche methods were approximately equal to consensus U-ALB, Abbott 4 % above, and Siemens 5 % below. For POCT, HemoCue was 1 % below, Siemens 7 % above, and Abbott 8 % below. For U-Creatinine, all manufacturers generally had a good agreement differing on average by 1–4 % from consensus. ConclusionsAlthough U-ALB methods generally meet The National Kidney Disease Education Program (NKDEP) recommendations of method bias less than 13 % and imprecision less than 30 %, differences among manufacturers have increased over the last years, with 2023 showing the largest differences between methods. This highlights the need for guidelines for albuminuria and ACR to take method differences into consideration, but also for implementation of suitable urine reference materials.

Extent of Equivalence of Results for Urine Albumin among 3 Candidate Mass Spectrometry Reference Measurement Procedures.

Urine albumin (UA) is an important biomarker of chronic kidney disease. Current in vitro diagnostic medical devices (IVD-MDs) for measuring UA are not standardized, and median results among IVD-MDs differ by approximately 45%. Since fixed decision values are used to interpret UA, higher-order reference measurement procedures (RMPs) are needed for metrological traceability. Three candidate liquid chromatography-tandem mass spectrometry RMPs have been developed for UA. Eight single-donation human urine samples were measured by 3 candidate RMPs. Results were compared using t-test and variance component analysis. The mean results for each urine sample from each RMP laboratory were not statistically different from the overall mean value by t-test. The median total CV including contributions from bias and imprecision among the 3 RMP laboratories was 6.23% using variance component analysis for each sample. The allowable bias to the RMP for an end-user IVD-MD was ≦9.0% or ≦3.0% based on the desirable or optimal total allowable error of 30% or 24%, respectively. A maximum allowable standard uncertainty for an RMP result was determined to be 4.3% or 3.3% for desirable or optimal performance, respectively. The standard uncertainties for all of the RMP laboratories meet the desirable and optimal standard uncertainty specifications. The candidate RMPs for UA in these 3 laboratories have suitable agreement of results and uncertainties for use as higher-order RMPs in the metrological traceability of end-user IVD-MDs for measuring UA.

Analytical performance evaluation of the GreenCare A1c and Cera-Stat HbA1c point-of-care testing assays.

The escalating prevalence of diabetes underscores the need for precise diagnostic tools to facilitate effective management. Hemoglobin A1c (HbA1c) is a crucial biomarker for long-term glycemic control in diabetic patients. Point-of-care testing (POCT) for HbA1c offers rapid, accessible alternatives to conventional laboratory methods, but uncertainties persist regarding the accuracy and reliability of POCT assays. This study evaluates the analytical performance of two boronate-affinity based HbA1c POCT assays, the GreenCare A1c and Cera-Stat HbA1c. Various analytical parameters including precision, linearity, comparison, and accuracy are assessed following guidelines from Clinical and Laboratory Standards Institute (CLSI), with results applied to certification criteria from the National Glycohemoglobin Standardization Program (NGSP) and International Federation of Clinical Chemistry and Laboratory Medicine (IFCC). Furthermore, 52 and 13 frozen EDTA whole blood samples were respectively used for additional evaluation of accuracy and interference due to Hb variants for the GreenCare A1c assay. Both GreenCare and Cera-Stat demonstrated good precision (repeatability CV% 1.5-1.9 and total imprecision CV% 1.6-2.2), linearity (R2 = 0.9996 & 0.9990), and correlation (r = 0.982 & 0.978) with an established HbA1c analyzer, the Bio-Rad D100. The GreenCare also exhibited good accuracy with frozen EDTA samples with known HbA1c values. Both assays met the certification criteria from NGSP and IFCC, classifying them as "standard" according to IFCC model for quality targets for HbA1c. This evaluation affirms the reliability of GreenCare and Cera-Stat POCT assays for HbA1c measurements, which can potentially reduce unnecessary referrals and enhance the overall quality of diabetes diagnosis and treatment.

Risk of mortality associated to chronic kidney disease in patients with complete left bundle branch block

Chronic kidney disease (CKD) is associated with cardiac conduction defects and is a strong risk factor for heart failure. Complete left bundle branch block (cLBBB), a cardiac conduction abnormality, may have an unfavorable effect on ventricular mechanical synchrony and lead to the progression of heart failure. Once heart failure develops, it seems to act together with underlying CKD in a vicious circle. Therefore, this study aimed to explore the influence of CKD in patients with cLBBB by assessing the estimated glomerular filtration rate (eGFR). We examined a hospital-based sample of 416 adult patients with cLBBB from 2010 to 2013. The eGFR was calculated using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation. Cox proportional hazard models were used to estimate the hazard ratio for all-cause mortality and cardiovascular mortality. A total of 416 adult patients with a mean age of 71 ± 13 years were enrolled. The median follow-up period was 3.6 years. After adjusting for clinical, electrocardiographic parameters, and medication use, cox regression analysis showed that total mortality was significantly associated with older age (Hazard Ratio (HR) = 1.03, 95% CI = 1.01–1.05, p = 0.002), presence of congestive heart failure (HR = 2.39, 95% CI = 1.63–3.49, p < 0.001), advanced CKD (HR = 2.48, 95% CI = 1.71–3.59, p < 0.001), higher HR (HR = 1.02, 95% CI = 1.01–1.03, p < 0.001) and without use of ACEI/ARB (HR = 0.59, 95% CI = 0.41–0.85, p = 0.005) were independent predictors of the total mortality. Multivariate Cox hazard regression analysis demonstrated that, in comparison to patients lacking cLBBB, the coexistence of CKD (eGFR < 60 mL/min/1.73 m2) among those with LBBB significantly heightened the risks of both total mortality (HR ratio of 5.01 vs. 2.40) and CV death (HR ratio of 61.78 vs. 14.41) even following adjustment for clinical covariates and ECG parameters. In summary, within patients exhibiting cLBBB, the presence of CKD serves as a significant risk factor for all-cause mortality.

Cardiovascular-kidney-metabolic overlap in heart failure with preserved ejection fraction: Cardiac structure and function, clinical outcomes, and response to sacubitril/valsartan in PARAGON-HF.

Cardiovascular-kidney-metabolic (CKM) multimorbidity is prevalent among individuals with heart failure (HF), but whether cardiac structure and function, clinical outcomes, and treatment response to sacubitril/valsartan vary in relation to CKM status is unknown. In this PARAGON-HF post-hoc analysis, we evaluated the impact of CKM multimorbidity (atherosclerotic cardiovascular [CV] disease, chronic kidney disease, and type 2 diabetes) on cardiac structure and function, clinical outcomes, and treatment effects of sacubitril/valsartan versus valsartan. The primary outcome was a composite of total HF hospitalizations and CV death. Secondary outcomes included the individual components of the primary outcome and a composite kidney outcome (sustained estimated glomerular filtration rate reduction of ≥50%, end-stage kidney disease, or kidney-related death). At baseline, 35.2% had one CKM condition, 33.3% had two, 15.9% had three, and only 15.6% had HF alone. CKM multimorbidity was associated with higher septal and posterior wall thickness, lower global longitudinal strain, higher E/e', and worse right ventricular function. Total HF hospitalizations or CV death increased with greater CKM multimorbidity, with the highest relative risk observed with three CKM conditions (rate ratio 3.06, 95% confidence interval 2.33-4.03), compared with HF alone. Treatment effects of sacubitril/valsartan were consistent irrespective of the number of CKM conditions for the primary endpoint (pinteraction = 0.75), CV death (pinteraction = 0.82), total HF hospitalizations (pinteraction = 0.67), and the composite kidney endpoint (pinteraction = 0.99). Cardiovascular-kidney-metabolic multimorbidity was common in PARAGON-HF and associated with adverse changes in cardiac structure and function and with a stepwise increase in risk of clinical outcomes. Treatment effects of sacubitril/valsartan were consistent irrespective of CKM burden. ClinicalTrials.gov NCT01920711.

End-user verification results of two serum separator tubes for clinical chemistry analytes according to CLSI GP34-A and CLSI GP41-A6

Tube manufacturers use different composition of gels and blood clot activator formulations in serum tube production. Our aim was to investigate the within-tube (repeatability) and between-tube variation, concordance between comparison results of BD and VacuSEL tubes. Blood samples were collected from control subjects (n = 20) and patients (n = 30) in accordance with the CLSI GP41-A6 and CLSI GP34-A guidelines. Twenty-three clinical chemistry parameters were analysed via Roche Cobas C702 Chemistry Analyzer on T0 (0 hour) and T24 (24 hour). Mean differences % were compared with Wilcoxon matched pair test. Clinical significance was evaluated based on desirable bias according to total allowable error (TEa). VacuSEL tubes demonstrated acceptable performance for the results of 20 parameters with regards to desirable bias % limits. Lactate dehydrogenase (LD) [mean difference % (%95 confidence intervals (CI) values of BD and VacuSEL tubes at T0 [6.41% (4.80–8.01%)]; sodium (Na) and total protein (TP) at T24 [-0.27% (-0.46 to −0.07%) and −1.39% (-1.87 to −0.91), respectively] were over the desirable bias limits (LD: 4.3%, Na: 0.23% and TP: 1.36%, respectively) but not exceeding total biological variation CV % [Na: 0.5 (0.0–1.0) % and TP: 2.6 (2.3–2.7) %). %95 confidence intervals (CI) of T0 LD values overlap with within-subject biological variation % (CI) limits (LD: 5.2 (4.9–5.4) %). The differences between two tubes were not medically significant and necessarily conclusive. VacuSEL serum tubes presented comparable performance with BD serum tubes.

GENDER DIFFERENCES IN ARTERIAL STIFFNESS IN YOUNG PATIENTS WITH PRIMARY HYPERTENSION

Objective: Influence of sex hormones on regulation of arterial stiffness (AS) include different mechanisms where changes in extracellular matrix, smooth and endothelial cells, and influence on oxidative stress and chronic inflammation are highlighted. In women after puberty AS decreases what confirm selective and fluctuating effect of sex hormones, primarly estrogen. In older men, hypogonadism is associated with incresed AS, while testosterone replacement reduce pulse wave velocity (PWV). Our aim was to analyze difference in central hemodynamic parameters between men and women with stage 1 hypertension. Design and method: This study included 154 patients with the primary hypertnsion (69.5% women, mean age 38.5). In all of them we collected different data and antroplogical paramaters using medical records. Parameters of AS were measured with the office tonometric system (SphygmoCor) and 24h ambulatory blood pressure monitoring system using Mobil-o-Graph and Arteriograph, but also estimated PWV was calculated using validated equation (ePWV). Results: There were no statistically significant gender differences in PWV values measured in the office oCor, measured during 24 hours (using Arteriograph and Mobil-O-Graph), and ePWV. There were no statistically significant gender differences in the values of 24 hours systolic and diastolic central aortic pressure (CAP) (Arteriograph or Mobil-O-Graph). Women had significant higher office CAP Men, on average, had statistically significantly higher amplified pulse pressure (APP) values than women (144.20 vs. 128.94, t = 3.275, p &lt; 0.01), while women had statistically significantly higher office augumentation index (AiX) values (t = -6.118, p &lt; 0.001), as well 24 hours (Mobil- O-Graph (t = -5.940, p &lt; 0.001;Arteriograph (t = -5.580, p &lt; 0.001). Conclusions: Possible explanation of no statistically significant differences in PWV values between men and women is average age of our group. A possible gender difference in AiX can be consequence of the increased reflection of pulse waves, which is increased in women. Longitudinal study should be conducted with monitoring of patients at longer time intervals in order to determine the further hormonal influence on structure of the arteries and for the assesment of total CV risk.

Body mass index and cardiorenal outcomes in the EMPEROR-Preserved trial: Principal findings and meta-analysis with the DELIVER trial.

Both low and high body mass index (BMI) are associated with poor heart failure outcomes. Whether BMI modifies benefits of sodium-glucose cotransporter 2 inhibitors (SGLT2i) in heart failure with preserved ejection fraction (HFpEF) requires further investigation. Using EMPEROR-Preserved data, the effects of empagliflozin versus placebo on the risks for the primary outcome (hospitalization for heart failure [HHF] or cardiovascular [CV] death), change in estimated glomerular filtration rate (eGFR) slopes, change in Kansas City Cardiomyopathy Questionnaire clinical summary score (KCCQ-CSS), and secondary outcomes across baseline BMI categories (<25 kg/m2, 25 to <30 kg/m2, 30 to <35 kg/m2, 35 to <40 kg/m2 and ≥40 kg/m2) were examined, and a meta-analysis conducted with DELIVER. Forty-five percent had a BMI of ≥30 kg/m2. For the primary outcome, there was a consistent treatment effect of empagliflozin versus placebo across the BMI categories with no formal interaction (p trend = 0.19) by BMI categories. There was also no difference in the effects on secondary outcomes including total HHF (p trend = 0.19), CV death (p trend = 0.20), or eGFR slope with slower declines with empagliflozin regardless of BMI (range 1.12-1.71 ml/min/1.73 m2 relative to placebo, p trend = 0.85 for interaction), though there was no overall impact on the composite renal endpoint. The difference in weight change between empagliflozin and placebo was -0.59, -1.48, -1.54, -0.87, and - 2.67 kg in the lowest to highest BMI categories (p trend = 0.016 for interaction). A meta-analysis of data from EMPEROR-Preserved and DELIVER showed a consistent effect of SGLT2i versus placebo across BMI categories for the outcome of HHF or CV death. There was a trend toward greater absolute KCCQ-CSS benefit at 32 weeks with empagliflozin at higher BMIs (p = 0.08). Empagliflozin treatment resulted in broadly consistent cardiac effects across the range of BMI in patients with HFpEF. SGLT2i treatment yields benefit in patients with HFpEF regardless of baseline BMI.

Analytical validation of the DropXpert S6 system for diagnosis of chronic myelocytic leukemia.

Digital PCR is a powerful method for absolute nucleic acid quantification and is widely used in the absolute quantification of viral copy numbers, tumor marker detection, and prenatal diagnosis. However, for most of the existing droplet-based dPCR systems, the droplet generation, PCR reaction, and droplet detection are performed separately using different instruments. Making digital PCR both easy to use and practical by integrating the qPCR workflow into a superior all-in-one walkaway solution is one of the core ideas. A new innovative and integrated digital droplet PCR platform was developed that utilizes cutting-edge microfluidics to integrate dPCR workflows onto a single consumable chip. This makes previously complex workflows fast and simple; the whole process of droplet generation, PCR amplification, and droplet detection is completed on one chip, which meets the clinical requirement of "sample in, result out". It provides high multiplexing capabilities and strong sensitivity while all measurements were within the 95% confidence interval. This study is the first validation of the DropXpert S6 system and focuses primarily on verifying its reliability, repeatability, and consistency. In addition, the accuracy, detection limit, linearity, and precision of the system were evaluated after sample collection. Among them, the accuracy assessment by calculating the absolute bias of each target gene yielded a range from -0.1 to 0.08, all within ±0.5 logarithmic orders of magnitude; the LOB for the assay was set at 0, and the LoD value calculated using probit curves is MR4.7 (0.002%); the linearity evaluation showed that the R2 value of the BCR-ABL was 0.9996, and the R2 value of the ABL metrics calculated using the ERM standard was 0.9999; and the precision evaluation showed that all samples had a CV of less than 4% for intra-day, inter-day, and inter-instrument variation. The CV of inter-batch variation was less than 7%. The total CV was less than 5%. The results of the study demonstrate that dd-PCR can be applied to molecular detection and the clinical evaluation of CML patients and provide more precise personal treatment guidance, and its reproducibility predicts the future development of a wide range of clinical applications.

Abstract 13724: Multiparameter Right Ventricular Dysfunction Predicts Worse Prognosis in Heart Failure With Preserved Ejection Fraction - Lessons From the PARAGON-HF Trial

Introduction: Impaired tricuspid annular plane systolic excursion (TAPSE), fractional area change (FAC) and right ventricular free wall longitudinal strain (RVFWLS) have been individually associated with a worse prognosis in patients with heart failure with preserved ejection fraction (HFpEF). However, these parameters measure different aspects of RV function and understanding their cumulative effect may provide insights into the overall impact of RV dysfunction (RVD) in HFpEF patients. Goal: Assess the association between the number of echocardiographic RVD parameters and clinical outcomes. Methods: Patients from the echocardiographic sub-study of the PARAGON-HF trial with adequate image quality (N=809, 53% women, mean age 74±8 years) were categorized based on the number of measures of RVD, which included TAPSE &lt;1.7 cm, FAC &lt;35% or RVFWLS &lt;20% (absolute value). Associations with total HF hospitalizations (HFH) and CV death, total HFH alone, first HFH or CV death, all-cause death and CV death were assessed by Groups, after accounting for confounders. Results: A total of 348 patients (43%) in Group 0 (no abnormal parameter), 355 (44%) in Group 1 (one abnormal parameter) and 106 (13%) in Group 2 (two or more abnormal parameters) were identified. After multivariable adjustment ( Table ), Group 2 was significantly associated with an increased risk of total HFH and CV death (RR 2.13, 95%CI 1.13-4.01), first HFH or CV death (RR 2.07, 95%CI 1.15-3.73), all-cause death (RR 2.61, 95%CI 1.19-5.74) and CV death (RR 3.08, 95%CI 1.14-8.31). The treatment effect of sacubitril/valsartan for total HFH and CV death was not modified by RVD Group (p-value for interaction=0.94). Conclusions: The presence of two or more RVD parameters was associated with a significantly worse prognosis, while an isolated RVD measure was not. These findings suggest that a more comprehensive approach to assessment of RV function may have better prognostic value in HFpEF.

Sex and age differences of major cardiovascular events in patients after percutaneous coronary intervention.

Women usually have higher risk after receiving percutaneous coronary interventions (PCIs) than men with coronary artery disease (CAD). The aim of this study was to investigate the association of sex differences with future outcomes in CAD patients undergoing PCI, to assess the role of age, and to extend observed endpoints to stroke and congestive heart failure. Six thousand six hundred forty-seven patients with CAD who received successful PCIs. The associations between clinic outcomes and sex were analyzed. The primary outcome was major cardiovascular events (MACE), including cardiac death, nonfatal myocardial infraction, and nonfatal stroke. The secondary outcome was MACE and hospitalization for heart failure (total CV events). During a mean of 52.7 months of follow-up, 4833 men and 1614 women received PCI. Univariate and multivariate analyses showed that women were independently associated with an increased risk of cardiac death (HR, 1.78; 95% CI, 1.32-2.41), hospitalization for heart failure (HR, 1.53; 95% CI, 1.23-1.89), MACE (HR, 1.34; 95% CI, 1.10-1.63), and total CV events (HR, 1.39; 95% CI, 1.20-1.62). In the subgroup analysis, women aged under 60 years had higher cardiovascular risks than men of the same age category. Women with CAD after successful PCI had poorer cardiovascular outcomes than men. Additionally, younger women (aged <60 years) were especially associated with a higher risk of developing future adverse cardiovascular outcomes.

A-102 Development and Validation of Turbidimetric Retinol Binding Protein (RBP) Assay

Abstract Background Retinol-binding protein (RBP) is a non-glycosylated protein synthesized by the liver. The role of RBP is to transport retinol (vitamin A) from the liver to the target tissues. RBP is also a sensitive marker of undernutrition. Serum/plasma RBP levels decrease with Vitamin A deficiency, hepatocellular insufficiency, and severe inflammation. Serum/plasma RBP levels increase with renal failure, type-2 diabetes, and steatosis. Methods The immunoassay was developed and validated on the turbidimetric instrument Cobas c501 from Roche Diagnostics. The antibodies against RBP in the assay react with RBP in the sample, and the turbidity formed by formation of antibody-antigen complexes is measured at 340 nm and 700 nm. The analytical sensitivity was tested in a study including 4 samples, analysed in 3 replicates over 3 days using a protocol based on the CLSI guideline EP17. The limit of quantification (LoQ) is defined as the lowest concentration of an analyte that can be reliably detected and at which the total error meets the requirements for accuracy. Interference with triglycerides, haemoglobin and bilirubin was tested using a protocol based on the CLSI guideline EP07. Precision of the assay was tested in a 22-day precision study using a protocol based on the CLSI guideline EP05. 3 controls were measured 2 times a day in 2 replicates each time. Results The assay has a calibration range from 7.5 to 120 mg/L. Samples with higher concentrations are automatically diluted and reanalysed. No antigen excess was observed up to 600 mg/L. Precision achieved a total CV of &amp;lt;4 % for samples with concentrations in the range 34–63 mg/L. Analytical sensitivity of RBP was measured as 7.81 mg/L with a total error (TE) of ≤20%. No clinically relevant interference towards triglycerides (13 g/L), haemoglobin (6.9 g/L) and bilirubin (0.3 g/L) was observed. Conclusion The Gentian RBP assay is a turbidimetric assay based on the formation of antibody-antigen complexes and can be used for assessment of undernutrition, vitamin A deficiency, hepatocellular insufficiency, severe inflammation, renal failure, type-2 diabetes, and steatosis. The assay can be installed on high-throughput platforms from different manufacturers and provides accurate, cost efficient and fast results.

Interlaboratory variation for NT-proBNP among Swedish laboratories in an external quality program 2011-2021.

NT-proBNP is frequently used for ruling out heart failure. Different cut-offs are used depending on the clinical context, e.g. an acute or chronic condition. Medical decision limits have been suggested at 125 and 300ng/L or 400ng/L in international guidelines. However, there is limited standardization between NT-proBNP methods and using the same blood sample might cause different treatment of patients. Data from the external quality assessment program for NT-proBNP from Equalis, Sweden, were extracted for the period 2011-2021, and categorized according to manufacturer. Manufacturer median NT-proBNP values were compared to total median values. CV% was calculated for each manufacturer and in comparison to different levels of NT-proBNP. Roche was the most common method, and its median results were closest to the median consensus results. When looking at the total CV at NT-proBNP levels in the range of 0-500ng/L, the total CV varied from 4 to 27%. During 2019-2021, Siemens (Immulite, Centaur, Atellica) yielded results 16-20% above the consensus median depending on sample level. Similarly, Abbott was 5-7% above, while Roche and Siemens Stratus were 1% respectively 6-10% below the consensus median. The introduction of new manufacturers and methods in 2017 have caused the agreement between manufacturers to decline. This highlights the need for a common calibrator and reference materials, particularly since medical decision limits in guidelines, e.g. European Society of Cardiology 2021, which are mostly based on Roche methods, do not take these method differences into account.

Ultra-processed foods and mortality: analysis from the Prospective Urban and Rural Epidemiology study

BackgroundHigher intake of ultra-processed foods (UPFs) has been associated with increased risk of CVD and mortality in observational studies from Western countries but data from non-Western countries are limited. ObjectivesWe aimed to assess the association between consumption of UPFs and risk of mortality and major CVD in a cohort from multiple world regions. DesignThis analysis includes 138,076 participants without a history of CVD between the ages of 35 and 70 y living on 5 continents, with a median follow-up of 10.2 y. We used country-specific validated food-frequency questionnaires to determine individuals’ food intake. We classified foods and beverages based on the NOVA classification into UPFs. The primary outcome was total mortality (CV and non-CV mortality) and secondary outcomes were incident major cardiovascular events. We calculated hazard ratios using multivariable Cox frailty models and evaluated the association of UPFs with total mortality, CV mortality, non-CV mortality, and major CVD events. ResultsIn this study, 9227 deaths and 7934 major cardiovascular events were recorded during the follow-up period. We found a diet high in UPFs (≥2 servings/d compared with 0 intake) was associated with higher risk of mortality (HR: 1.28; 95% CI: 1.15, 1.42; P-trend < 0.001), CV mortality (HR: 1.17; 95% CI: 0.98, 1.41; P-trend = 0.04), and non-CV mortality (HR: 1.32; 95% CI 1.17, 1.50; P-trend < 0.001). We did not find a significant association between UPF intake and risk of major CVD. ConclusionsA diet with a high intake of UPFs was associated with a higher risk of mortality in a diverse multinational study. Globally, limiting the consumption of UPFs should be encouraged.

Total serum vitamin B12 (cobalamin) LC-MS/MS assay as an arbiter of clinically discordant immunoassay results.

Measurement of the serum levels of vitamin B12 (VB12) is key for evaluating VB12 deficiency-dependent anemia. Immunoassay, the major method for determining VB12, tends to give false-normal results because of the presence of anti-intrinsic factor (IF-Ab) or other factors such as heterophilic antibodies etal. This study aimed to develop a liquid chromatography tandem mass spectrometry (LC-MS/MS) method that is helpful for distinguish false normal VB12 results measured by the immunoassay. Different forms of VB12 were derivatized into CN-B12, which was collected through solid-phase extraction and analyzed via LC-MS/MS. 236 serum samples were measured both by LC-MS/MS and immunoassay, results were compared, and the IF-Ab effect was evaluated. The LC-MS/MS assay afforded a linear slope from 20 to 4,000pmol/L for CN-B12. OH-VB12, methyl-VB12, and CoA-VB12 showed recovery within 89.3-109.5%. The intra-assay CV of VB12 was 2.6-4.1%, whereas the total CV was 9.3-9.8%. Passing-Bablok regression between LC-MS/MS and immunoassay results showed that the slope was 1.085 and the intercept was-15.691. The Bland-Altman plot showed that the mean difference and difference% were-34.6 pmol/L and 0.3%, respectively. Inter-rater agreement analysis showed that the linear weighted kappa value was 0.885, implying good agreement between the two methods. However, two samples were falsely elevated and one sample was falsely normal in the immunoassay compared with LC-MS/MS. The LC-MS/MS method helped in the distinction of false-normal VB12 results shown by the immunoassay. The VB12 LC-MS/MS method can be used as an arbiter of clinically discordant immunoassay results.

A novel point-of-care device accurately measures thyrotropin in whole blood, capillary blood and serum.

Point-of-care (POC) measurement of thyrotropin (TSH) may facilitate prompt diagnosis of thyroid dysfunction. We evaluated the analytical performance of a new POC TSH assay (Wondfo). TSH measurements were made from 730 consecutive, unselected subjects in an outpatient setting, using Wondfo in whole blood, capillary blood and serum or automated reference equipment (serum only). TSH measurements were user-independent. Total intra-and inter-assay variation (CV%) was 12.1 and 16.2%, respectively. Total CV% was 10.6-22.6% and 14.5-21.6% in serum and whole blood, respectively. Linearity was very good. Recovery rate was 97-127%. Prolongation of incubation time increased TSH results of 12% (13%) and 33% (35%) after 2 and 5 additional minutes in serum (blood), respectively. When measured simultaneously in two Wondfo devices, the slope of the regression line was 1.03 (serum) and 1.02 (blood), with Spearman's correlation of 0.99 for both. TSH measurements between Wondfo and reference correlated strongly (r=0.93-0.96), though TSH measurements were lower with Wondfo (slopes of plots of measurements made using the two devices were 0.94 [serum vs. serum]; 0.83 [whole blood vs. serum] and 0.64 [capillary blood vs. serum]). Depending on sample material, TSH in capillary blood was lower vs. whole blood (slope: 0.82) and for whole blood vs. serum (Wondfo and reference method; slope: 0.69 and 0.83). Total haemolysis, but not elevated bilirubin or lipemia, disrupted TSH measurement. The Wondfo system was straightforward to use without need for specialist technicians and demonstrated analytic performance suitable for clinical use for the diagnosis of thyroid dysfunction.

PROGNOSTIC OF PATIENTS WITH REFRACTORY HYPERTENSION IN A COHORT OF RESISTANT HYPERTENSIVES

Objective: To determine prognostic value of true refractory hypertension diagnosis in a large cohort of resistant hypertensives Design and method: Prospective cohort study enrolled 835 patients of a large historical cohort of RHTN. After at least 6 months of follow-up, 147 (17.6%) patients remained refractory to treatment in ABPM despite the use of 5 or more drugs, including a thiazide-like diuretic and spironolactone. All of them were submitted to a standard protocol including laboratory tests, ABPM, echocardiography, and pulse wave velocity (PWV). Primary outcomes were total mortality, cardiovascular mortality, and fatal and non-fatal cardiovascular events. Secondary outcomes were major cardiovascular events separately (coronary artery disease, cerebrovascular disease, chronic kidney disease with substitutive therapy, peripheral arterial disease). Person-time rates for interest outcomes were calculated to find correlations of Kaplan-Meier survival curves using Cox regression multivariate models, after adjustments for several risk factors. Follow-up remained until December 2021 Results: The median follow-up was 162 months [IQR 115–196]. When compared to resistant hypertensives, patients with RfHTN were younger and more obese. Regarding subclinical lesions, refractories presented less aortic stiffness (13 vs 21%, p = 0.02) and stage 3 chronic kidney diseases (26 vs 34%, p = 0.03) with albuminuria and LVMI similar to those of resistant hypertension despite uncontrolled ambulatory blood pressure. There were 232 all causes deaths (21.1 vs 29.2%), of which 122 from CV causes (12.9% vs 15%), and 253 total CV events (44.1% vs 49%). The diagnosis of RfHTN was associated with a significantly lower risk of all-cause mortality when adjusted by sex and age. It also presented a tendency to be a protective factor of cardiovascular death, chronic kidney disease with substitutive therapy and peripheral arterial disease, although without statistical significance. Risk ratio ranged from 0.62 to 0,94 for every analysis/adjustment, except incidence of stroke (IC95%:1.22(0.70–2.17) after every adjustment in RfHTN. Conclusions: Despite uncontrolled ABPM, refractory hypertensive patients presented lower overall and cardiovascular morbimortality at long-term when compared to non-refractory ones, except stroke, which seems to be at least partially influenced by spironolactone use.

Sodium-glucose cotransporter-2 inhibitors in heart failure: an updated meta-analysis.

AimsWe aimed to examine efficacy and safety outcomes of sodium‐glucose cotransporter‐2 inhibitor (SGLT2i) for the treatment of heart failure (HF), especially in patients with heart failure with preserved ejection fraction (HFpEF).Methods and resultsPubMed, Web of Science, and Cochrane Library were searched to identify randomized controlled trials comparing SGLT2i vs. placebo in HF patients. A total of 10 studies with 23 852 HF patients were eventually included. Compared with placebo, SGLT2i is associated with a lower incidence of composite of first hospitalization for heart failure (HHF) or cardiovascular death (CV death) [hazard ratio (HR) = 0.76 95% confidence interval (CI) = 0.71–0.81], which is consistent regardless of the diabetes status, type of gliflozines used, and follow‐up duration. SGLT2i can reduce the risk of total HHF or CV death (HR = 0.74, 95%CI = 0.68–0.81), first HHF (HR = 0.69, 95%CI = 0.64–0.75), CV death (HR = 0.88, 95%CI = 0.80–0.96), any death (HR = 0.90, 95%CI = 0.83–0.97), and any serious events (HR = 0.90, 95%CI = 0.87–0.93) in HF patients, at the cost of increased risk of urinary tract infections (risk ratio = 1.17, 95%CI = 1.03–1.33). In HFpEF patients, SGLT2i is associated with a significant reduction of composite of first HHF or CV death (HR = 0.81, 95%CI = 0.73–0.91), first HHF (HR = 0.71, 95%CI = 0.62–0.82), and total HHF or CV death (HR = 0.61, 95%CI = 0.43–0.86).ConclusionsSodium‐glucose cotransporter‐2 inhibitor contributed to better efficacy outcomes in overall HF patients and showed an inspiring breakthrough in the treatment of HFpEF.

Statin use in older people primary prevention on cardiovascular disease: an updated systematic review and meta-analysis.

Evidence on statin use for primary prevention of cardiovascular disease (CVD) in older people needs to be extended and updated, aiming to provide further guidance for clinical practice. PubMed, EMBASE, Cochrane Library and Web of Science were searched for eligible observational studies comparing statin use vs. no-statin use for primary prevention of CVD in older people (age 65 years). The primary outcomes were all-cause mortality, CVD mortality, coronary heart disease (CHD)/myocardial infraction (MI), stroke and total CV events. Risk estimates of each relevant outcome were synthesized as a hazard ratio (HR) with 95% confidence interval (95% CI) using in the random-effects model. Twelve eligible observational studies (n = 1,627,434) were enrolled. The pooled results suggested that statin use was associated with a significantly decreased risk of all-cause mortality (HR: 0.54, 95% CI: 0.46-0.63), CVD mortality (HR: 0.51, 95% CI: 0.39-0.65), CHD/MI (HR: 0.83, 95% CI: 0.69-1.00), stroke (HR: 0.79, 95% CI: 0.68-0.92) and total CV events (HR: 0.75, 95% CI: 0.66-0.85). The association in all-cause mortality still remained obvious at higher ages ( 70 years old, HR: 0.56, 95% CI: 0.44-0.71; 75 years old, HR: 0.70, 95% CI: 0.60-0.80; 85 years old, HR: 0.85, 95% CI: 0.74-0.97), 20% (HR: 0.47, 95% CI: 0.35-0.62) and 20% diabetic populations (HR: 0.50, 95% CI: 0.40-0.64), and 50% (HR: 0.68, 95% CI: 0.59-0.79) and 50% hypertensive populations (HR: 0.38, 95% CI: 0.16-0.88). Statin use was related to a 46%, 49%, 17%, 21% and 25% risk reduction on all-cause mortality, CVD mortality, CHD/MI, stroke and total CV events in older patients, respectively. The significant association was also addressed in older patients and 75 years old individuals for CVD primary prevention.

Development and Provisional Validation of a Multiplex LC-MRM-MS Test for Timely Kidney Injury Detection in Urine.

Kidney injury is a complication frequently encountered in hospitalized patients. Early detection of kidney injury prior to loss of renal function is an unmet clinical need that should be targeted by a protein-based biomarker panel. In this study, we aim to quantitate urinary kidney injury biomarkers at the picomolar to nanomolar level by liquid chromatography coupled to tandem mass spectrometry in multiple reaction monitoring mode (LC-MRM-MS). Proteins were immunocaptured from urinary samples, denatured, reduced, alkylated, and digested into peptides before LC-MRM-MS analysis. Stable-isotope-labeled peptides functioned as internal standards, and biomarker concentrations were attained by an external calibration strategy. The method was evaluated for selectivity, carryover, matrix effects, linearity, and imprecision. The LC-MRM-MS method enabled the quantitation of KIM-1, NGAL, TIMP2, IGFBP7, CXCL9, nephrin, and SLC22A2 and the detection of TGF-β1, cubilin, and uromodulin. Two to three peptides were included per protein, and three transitions were monitored per peptide for analytical selectivity. The analytical carryover was <1%, and minimal urine matrix effects were observed by combining immunocapture and targeted LC-MRM-MS analysis. The average total CV of all quantifier peptides was 26%. The linear measurement range was determined per measurand and found to be 0.05–30 nmol/L. The targeted MS-based method enables the multiplex quantitation of low-abundance urinary kidney injury biomarkers for future clinical evaluation.