Planar Functionalized Surfaces for Direct Immunoaffinity Desorption/Ionization Mass Spectrometry.

BackgroundExcess levels of serum acylcarnitines, which are intermediate products in metabolism, have been observed in metabolic diseases such as type 2 diabetes mellitus. However, it is not known whether acylcarnitines may prospectively predict risk of cardiovascular death or acute myocardial infarction in patients with stable angina pectoris.Methods and ResultsThis study included 4164 patients (median age, 62 years; 72% men). Baseline serum acetyl‐, octanoyl‐, palmitoyl‐, propionyl‐, and (iso)valerylcarnitine were measured using liquid chromatography/tandem mass spectrometry. Hazard ratios (HRs) and 95% CIs for quartile 4 versus quartile 1 are reported. The multivariable model included age, sex, body mass index, fasting status, current smoking, diabetes mellitus, apolipoprotein A1, apolipoprotein B, creatinine, left ventricular ejection fraction, extent of coronary artery disease, study center, and intervention with folic acid or vitamin B6. During median 10.2 years of follow‐up, 10.0% of the patients died of cardiovascular disease and 12.8% suffered a fatal or nonfatal acute myocardial infarction. Higher levels of the even‐chained acetyl‐, octanoyl‐, and palmitoyl‐carnitines were significantly associated with elevated risk of cardiovascular death, also after multivariable adjustments (HR [95% CI]: 1.52 [1.12, 2.06]; P=0.007; 1.73 [1.23, 2.44]; P=0.002; and 1.61 [1.18, 2.21]; P=0.003, respectively), whereas their associations with acute myocardial infarction were less consistent.ConclusionsAmong patients with suspected stable angina pectoris, elevated serum even‐chained acylcarnitines were associated with increased risk of cardiovascular death and, to a lesser degree with acute myocardial infarction, independent of traditional risk factors.Clinical Trial RegistrationURL: http://www.clinicaltrials.gov. Unique identifier: NCT00354081.

Background Our previous research has identified the haptoglobin (Hp) phenotype as a potential biomarker that could be used to personalize glycemic control in type 2 diabetes mellitus (T2DM) to prevent coronary artery disease (CAD), the most common cardiovascular disease (CVD). We observed that attaining HbA1c ≥8.0% compared with 7.0–7.9% was consistently associated with incident CAD risk among participants in the Action to Control Cardiovascular Risk in Diabetes (ACCORD) study with the Hp2-2 type(1). No similar association was observed among participants without the Hp2-2 type. However, most ACCORD participants were middle-aged and older males who were at high risk of CVD. Therefore, the optimal glycemic target for CAD prevention for the Hp types remains unclear among women, younger adults, and those at lower risk of CVD. Objective To explore the relationship between achieving specific clinically relevant HbA1c targets and risk of incident CAD in separate Hp types in a diverse cohort of individuals living with T2DM (the Action for Health in Diabetes (Look AHEAD) study(2)). Methods Hp type was measured in 4,539 blood samples from the Look AHEAD study using a validated assay. Cox regression models with time-varying covariables were used to quantify the association between time-varying achieved HbA1c (<6.5%, 6.5-6.9% and ≥8.0% compared to 7.0-7.9%), updated at years 1-4 and biennially thereafter, and incident CAD in the Hp2-2 (n=1,590) and non-Hp2-2 (n=2,949) types separately. Further pre-specified subgroup analyses by age, sex, and history of CVD were performed in each Hp type group separately. Results During 16 years of follow-up, 744 incident CAD cases were documented. Compared with HbA1c 7.0-7.9%, having HbA1c <6.5% was associated with a 29% lower CAD risk among participants with the non-Hp2-2 type (adjusted HR 0.71, 95% CI 0.55-0.90). In subgroup analyses, this association was present in participants with the non-Hp2-2 type who were male (0.58, 0.42-0.80), who did not have a history of CVD (0.67, 0.49-0.93), and who were aged ≥65 years at baseline (0.64, 0.43-0.95). HbA1c ≥8.0% was associated with CAD risk among participants with the Hp2-2 type who had a history of CVD (1.81, 1.02-3.21). No associations were observed between the other HbA1c targets and CAD risk when Hp2-2 participants were combined or partitioned into subgroups. Conclusion Achieving HbA1c <6.5% compared with 7.0–7.9% reduced the risk of CAD events in Look AHEAD participants with the non-Hp2-2 type, but not in participants with the Hp2-2 type. HbA1c ≥8.0% was associated with higher CAD risk among participants who had the Hp2-2 type and a history of CVD.

Myocardial Infarction in Heart Failure With Preserved Ejection Fraction: Pooled Analysis of 3 Clinical Trials

Background: The aim of this paper is to assess the impact of sodium-glucose cotransporters-2 (SGLT2) inhibitors on all-cause and cardiovascular (CV) death in high-risk patients and compare the efficacy of empagliflozin and dapagliflozin. Methods: PubMed was queried from inception to the last week of September 2023 for randomized controlled trials that compared SGLT2 inhibitors’ empagliflozin or dapagliflozin with placebo and included patients with heart failure (HF), type 2 diabetes mellitus (T2DM), or chronic kidney disease (CKD). The outcome of interest was CV death or all-cause death. Hazard ratios (HR) with 95% confidence intervals (CI) were pooled using a random effect model, and forest plots were created to analyze the results visually. A chi-square test was performed to assess subgroup differences between empagliflozin and dapagliflozin. Results: Eight trials (N=55,818) were included in our analysis, namely EMPA-REG, EMPEROR-Reduced, EMPEROR-Preserved, EMPA-KIDNEY, DECLARE-TIMI, DAPA-HF, DELIVER and DAPA-CKD. Pooled analysis demonstrated that compared to placebo, SGLT2 inhibition reduced the risk of CV death (SGLT2i arm = 1405 events, 29,089 total patients; placebo arm = 1515 events, 26,729 total patients; HR: 0.85; 95%CI: 0.79-0.93, p<0.001) and all-cause death (SGLT2i arm = 2,491 events, 29,062 total patients; placebo arm= 2,625 events, 26,729 total patients; HR: 0.86; 95% CI 0.79-0.95, p=0.002) in high-risk patients identified as having either T2DM, HF, or CKD. No differences were observed in the effect of empagliflozin and dapagliflozin on CV death (HRempagliflozin: 0.81; 95% CI 0.68-0.97, HRdapagliflozin: 0.88; 0.82-0.95, p=0.39) and all-cause death (HRempagliflozin: 0.86; 95% CI 0.73-1.02, HRdapagliflozin: 0.87; 0.78-0.97, p=0.94). Conclusions: SGLT2 inhibitors reduce the risk of all-cause and CV death in high-risk patients. Notably, there were no discernible differences in the benefits of empagliflozin and dapagliflozin on these outcomes.

Background:To investigate the efficacy and safety of sodium-glucose cotransporter 2 (SGLT-2) inhibitors in patients with heart failure (HF) and type 2 diabetes mellitus (T2DM).Methods:A manual search was conducted in 3 prestigious English databases, Cochrane Library, PubMed, and Web of Science, for studies published within the last decade, from July 2014 to July 2024. The extracted literature was synthesized to analyze the efficacy outcomes, survival prognostic indicators, and safety profiles of SGLT-2 inhibitors in patients with HF and T2DM. The Cochrane bias risk assessment scale was used as a tool to evaluate the quality of the literature, and Review Manager 5.4 software was used to create the bias risk chart. Data analysis and merging were completed with the help of Review Manager 5.4 and Stata 15.0 statistical software.Results:Twelve studies encompassing 9509 patients were included in the meta-analysis. The results revealed that compared to the control group, the SGLT-2 inhibitor-treated group demonstrated significantly greater reductions in left ventricular end-diastolic volume index [mean difference (MD) = -7.25, 95% confidence intervals [95% CI] (-9.83, -4.67)], brain natriuretic peptide levels [MD = -36.96, 95% CI (-63.51, -10.41)], and N-terminal pro-brain natriuretic peptide [MD = -519.27, 95% CI (-660.77, -377.78)]. Furthermore, the SGLT-2 inhibitor-treated group exhibited significantly higher increases in Kansas City Cardiomyopathy Questionnaire scores [MD = 3.32, 95% CI (3.30, 3.34)], indicating improved quality of life. Additionally, the incidence of adverse events was significantly lower in the SGLT-2 inhibitor-treated group compared to the control group [OR = 0.78, 95% CI (0.69, 0.88)]. The pooled results of the meta-analysis indicated that SGLT-2 inhibitor therapy reduced the risk of cardiovascular death or HF hospitalization by 23%, the risk of cardiovascular death by 19%, and the risk of all-cause mortality by 9%.Conclusion:SGLT-2 inhibitor therapy significantly reduced the risks of all-cause mortality, cardiovascular death, and hospitalization for HF in patients with HF and T2DM. Additionally, SGLT-2 inhibitors significantly improve cardiac function, decrease the incidence of adverse events, and enhance the quality of life in these patients.

Recognizing Pregnancy-Associated Cardiovascular Risk Factors

Plasma levels of Apolipoprotein A1 and Lecithin:Cholesterol Acyltransferase in type 2 diabetes mellitus: Correlations with haptoglobin phenotypes

Chronic kidney disease from screening, detection, and awareness, to prevention.

Haptoglobin is an acute-phase protein with three major phenotypes: Hp 1-1, Hp 2-1 and Hp 2-2. Studies have shown that functional differences between these phenotypes have important consequences in a number of pathological disorders (e.g. cardiovascular disease, autoimmune disorders, infectious disease), making phenotype determination of potential use in the clinical field. Previous methods for haptoglobin phenotyping have involved electrophoresis (starch, acetate and polyacrylamide gels) with phenotype visualization by peroxidase-sensitive stains that were often carcinogenic. Less hazardous immunoblotting and isoelectric focusing procedures are also available but the methods are lengthy, expensive and often complex. Here, we describe a new method for haptoglobin phenotyping using commercially available agarose gels and a non-carcinogenic stain (3,3',5,5'-tetramethylbenzidine). Our method showed 100% agreement with starch gel electrophoresis and reliably distinguished between commercial haptoglobin phenotype standards and serum samples from patients (n = 125). We obtained a sensitivity of 0.4 g/L for all phenotypes and the gels were stable for up to 2 months. This safer and easier to use method may permit increased access to knowledge of haptoglobin phenotype, allowing improved patient management and better tailoring of treatment in a variety of clinical conditions.

Intact-mass measurements are becoming increasingly popular in mass spectrometry (MS) based protein characterization, as they allow the entire complement of proteoforms to be evaluated within a relatively short time. However, applications of this approach are currently limited to systems exhibiting relatively modest degrees of structural diversity, as the high extent of heterogeneity frequently prevents straightforward MS measurements. Incorporation of limited charge reduction into electrospray ionization (ESI) MS is an elegant way to obtain meaningful information on most heterogeneous systems, yielding not only the average mass of the protein but also the mass range populated by the entire complement of proteoforms. Application of this approach to characterization of two different phenotypes of haptoglobin (1-1 and 2-1) provides evidence of a significant difference in their extent of glycosylation (with the glycan load of phenotype 2-1 being notably lighter) despite a significant overlap of their ionic signals. More detailed characterization of their glycosylation patterns is enabled by the recently introduced technique of cross-path reactive chromatography (XP-RC) with online MS detection, which combines chromatographic separation with in-line reduction of disulfide bonds to generate metastable haptoglobin subunits. Application of XP-RC to both haptoglobin phenotypes confirms that no modifications are present within their light chains and provides a wealth of information on glycosylation patterns of the heavy chains. N-Glycosylation patterns of both haptoglobin phenotypes were found to be consistent with bi- and triantennary structures of complex type that exhibit significant level of fucosylation and sialylation. However, multivariate analysis of haptoglobin 1-1 reveals higher number of the triantennary structures, in comparison to haptoglobin 2-1, as well as a higher extent of fucosylation. The glycosylation patterns deduced from the XP-RC/MS measurements are in agreement with the conclusions of the intact-mass analysis supplemented by limited charge reduction, suggesting that the latter technique can be employed in situations when fast assessment of protein heterogeneity is needed (e.g., process analytical technology applications).

Objective: Systolic blood pressure, prevalence of smoking, family history of coronary heart disease, cholesterol and cardiorespiratory fitness are well established as cardiovascular risk factors. In the present study we aimed to investigate how one measurement these variables at middle-age influenced risk of cardiovascular death during the first, intermediate and late part of a 35-year observation period, respectively.Design and method: Systolic blood pressure, prevalence of smoking, family history of coronary heart disease, cholesterol and cardiorespiratory fitness were measured in 2014 apparently healthy, middle-aged Caucasian men by clinical examination, blood tests and an ECG-monitored, symptom-limited bicycle test. End points were registered after 35 years in a nationwide scrutiny of charts in all hospitals, and from the national Cause of Death registry. Early cardiovascular death was defined as event before 12 years of observation, intermediate event between 12 and 23 years of observation and late event between 23 and 35 years of observation. Impact of predictors and relative risks were estimated using Cox proportional hazards models. When estimating risks of intermediate and late cardiovascular death, men who died within 12 and 23 years were excluded. Results: We found 80 events of early, 207 events of intermediate and 241 events of late cardiovascular death. Age, systolic blood pressure, smoking status, and cholesterol were significant predictors of early, intermediate and late cardiovascular death. Baseline physical fitness was a significant and independent predictor of early and a borderline significant predictor of intermediate cardiovascular death but had no predictive impact on late cardiovascular death. Family history of coronary heart disease was a significant and independent predictor of early and intermediate, but not late cardiovascular death (Table1). Conclusions: Systolic blood pressure, smoking, and cholesterol measured at a median age 49 years were strong predictors of early, intermediate and late cardiovascular death during 35 years. Physical fitness at middle age and family history of coronary heart disease were only associated with risk of cardiovascular death before reaching 72 years. Risk factors for CV death have different persistence throughout life and systolic blood pressure at middle-age remains a significant cardiovascular risk factor until old ages.

IntroductionIn patients with carotid stenosis receiving medical treatment, carotid plaque echolucency has been thought to predict risk of future stroke and of other cardiovascular events. This study evaluated the prognostic value of pre-operative plaque echolucency for future stroke and cardiovascular death in patients undergoing carotid endarterectomy in the first Asymptomatic Carotid Surgery Trial (ACST-1).MethodsIn ACST-1, 1832/3120 patients underwent carotid endarterectomy (CEA), of whom 894 had visual echolucency assessment according to the Gray-Weale classification. During follow-up patients were monitored both for peri-procedural (i.e. within 30 days) death, stroke, or MI, and for long-term risk of stroke or cardiovascular death. Unconditional maximum likelihood estimation was used to calculate odds ratios of peri-procedural risk and Kaplan-Meier statistics with log-rank test were used to compare cumulative long-term risks.ResultsOf 894 operated patients in whom echolucency was assessed, 458 plaques (51%) were rated as echolucent and peri-procedural risk of death/stroke/MI in these patients was non-significantly higher when compared with patients with non-echolucent plaques (OR 1.48 [95% CI 0.76–2.88], p = .241). No differences were found in the 10 year risk of any stroke (30/447 [11.6%] vs. 29/433 [11.0%], p = .900) or cardiovascular (non-stroke) death (85/447 [27.9%] vs. 93/433 [32.1%], p = .301).ConclusionIn ACST-1, carotid plaque echolucency assessment in patients undergoing CEA offered no predictive value with regard to peri-operative or long-term stroke risk or of cardiovascular (non-stroke) death.

Intact-mass measurements are becoming an increasingly popular in mass spectrometry (MS) based protein characterization, as they allow the entire complement of proteoforms to be evaluated within a relatively short time. However, applications of this approach are currently limited to systems exhibiting relatively modest degrees of structural diversity, as the high extent of heterogeneity frequently prevents straightforward MS measurements. Incorporation of limited charge reduction into electrospray ionization (ESI) MS measurements provides an elegant way to obtain meaningful information on most heterogeneous systems, yielding not only the average mass of the protein, but also the mass range populated by various proteoforms. Application of this approach to characterization of two different phenotypes of haptoglobin (1-1 and 2-1) provides evidence of a significant difference in their extent of glycosylation, with the glycan load of phenotype 2-1 being notably lighter. More detailed characterization of their glycosylation patterns is enabled by the recently introduced crosspath reactive chromatography (XP-RC) with on-line MS detection, a technique that combines chromatographic separation with in-line reduction of disulfide bonds to generate metastable haptoglobin subunits. Application of XP-RC to both haptoglobin phenotypes confirms that no modifications are present within their light chains, and provides a wealth of information on glycosylation patterns of the heavy chains. The haptoglobin 1-1 glycans are mature fully sialylated biantennary structures that exhibit high degrees of fucosylation. In contrast, phenotype 2-1 contains a significant fraction of incomplete biantennary structures and exhibit significantly lower levels of sialylation and fucosylation. The glycosylation patterns deduced from the XP-RC/MS measurements are in agreement with the conclusions of haptoglobin analysis by limited charge reduction, suggesting that the latter can be employed in situations when a fast assessment of a protein heterogeneity is needed (e.g., comparability studies of biopharmaceutical products).

Risk of myocardial infarction, ischemic stroke, and cardiovascular death in patients with atopic dermatitis

Background Type 2 diabetes mellitus (T2DM) is a major risk factor for cardiac diseases and renal dysfunction. Whether T2DM increases the risk of cardiorenal syndromes (CRS) subtypes to a similar extent, and whether the risk of deleterious outcomes after CRS is modified by diabetes are poorly known. Methods In a nationwide cohort study including 5,123,193 patients seen in French hospitals in 2012 with at least 5 years of follow-up (or dying earlier), we assessed the incidence of CRS subtypes, and then the impact of T2DM in patients with CRS on the risk of death, cardiovascular death, heart failure (HF), myocardial infarction (MI) and end-stage kidney disease (ESKD) during follow-up (27,735,205 person-years). Patients with history of dialysis, kidney transplantation or type 1 DM were excluded of the analysis. We performed 1:1 propensity matching on baseline characteristics including age, sex, risk factors, cardiovascular and non-cardiovascular comorbidities for patients with T2DM or no T2DM. The model by Fine and Gray was used for analyzing the competing risks for clinical events and all-cause death with sub-distribution hazard ratios (sHR). Results Among the 5,123,193 patients, 4,605,236 (91.2%) had neither HF nor CKD baseline. Among them, 391,186 (8.1%) had T2DM and 380,581 of them were matched 1:1 with 380,581patients with no T2DM. During follow-up, CRS occurred in 42,375 patients (incidence 0.98%/year): acute, i.e. type 1,3 or 5 CRS n=9,438, 22%; type 2 (cardiorenal) CRS n=21,075, 50%; type 4 (renocardiac) CRS n=11,862, 28%). In multivariable analysis, T2DM was the most powerful predictor of incident CRS (any type, HR: 2.182, 95% CI 2.150–2.214) among all baseline characteristics. The incidence of all-type CRS was higher in matched patients with T2DM (1.30%/year, 95% CI 1.29–1.32) than in those with no T2DM (0.65%/year, 95% CI 0.64–0.66): sHR 1.905 (95% CI 1.867–1.943). The risk of CRS associated with diabetes (vs no diabetes) was higher for type 4 (sHR 2.182, 95% CI 2.098–2.269) than for type 2 (sHR 1.834, 95% CI 1.783–1.887) and for acute (sHR 1.707, 95% CI 1.637–1.780) CRS. Among the 451,942 patients with HF or CKD at baseline, 26,396 patients had CRS at baseline, among whom 11,355 (43.0%) had diabetes: 8,314 of them were matched 1:1 with 8,314 with CRS and no T2DM. Compared to CRS patients with no diabetes, matched patients with CRS and T2DM had a greater incidence of all-cause death (sHR 1.085, 95% CI 1.048–1.123), cardiovascular death (sHR 1.145, 95% CI 1.080–1.214), ESKD (sHR 1.319, 95% CI 1.223–1.422), hospitalization for HF (sHR 1.119, 95% CI 1.078–1.162) and MI (sHR 1.294, 95% CI 1.139–1.470) during follow-up. Conclusions T2DM is a major risk factor for all CRS subtypes, may differently affect the incidence of type 2, type 4 and acute CRS and aggravates the risk of deleterious outcomes after CRS. Funding Acknowledgement Type of funding sources: None.