Abstract The Commission on Laboratory Diagnostics in Diabetology was founded in 2015 at the suggestion and with the support of the Executive Board of the German Diabetes Society (DDG) and the German Society of Clinical Chemistry and Laboratory Medicine (DGKL), with the aim of addressing methodological aspects for the reliable measurement of clinical chemistry measurands such as glucose, HbA 1c , insulin, C-peptide, antibodies, etc. This mini-review looks back on the work of the KLD over the past 10 years and briefly outlines current activities and future steps.

BACKGROUND:Cholesterol is a main contributor to coronary artery disease (CAD). Although the genetic basis of blood cholesterol concentration is well studied, there is currently a lack of studies investigating the genetics of its precursors from de novo biosynthesis. METHODS:We conducted a genome-wide association meta-analysis, combining data from KORA, LIFE-Heart, LIFE-Adult, LURIC, the Sorbs study, and YFS, resulting in up to 10,519 individuals. We investigated 14 traits related to serum concentrations of lanosterol, desmosterol, and cholesterol. Direct and indirect effects of lanosterol on CAD were investigated with a Mendelian randomisation mediation analysis. FINDINGS:Our analysis revealed four genome-wide significant (p < 5 × 10-8) associations not previously reported in the GWAS catalogue. These include two loci without prior connection to cholesterol, associated with lanosterol (7q21.2, CYP51A1) and free cholesterol (11q14.1), and two associations with lanosterol at loci previously reported for cholesterol (5q13.3, HMGCR; 11q23.3, APO cluster). We also replicated eight loci previously reported for associations with cholesterol-related traits. Lanosterol exhibited significant total and indirect effects on CAD, but its direct effect was not significant. INTERPRETATION:We demonstrate that the investigation of intermediate phenotypes can help to functionally fine map previously reported associations for cholesterol, improving our understanding of genetic regulation of cholesterol concentrations. Further, the effect of lanosterol on CAD is probably fully mediated by total cholesterol. FUNDING:This investigation was primarily funded by the ministry for science and health of the Rhineland-Palatinate through the CoAGE graduate programme. A complete list of funding organisations is provided in the acknowledgements.

BACKGROUND AND HYPOTHESIS:The endogenous opioid proenkephalin (PENK) has been identified as an independent risk marker for acute cardiovascular diseases and acute kidney injury. However, its regulation, its association with other cardiometabolic markers and its prognostic role in patients with chronic kidney disease (CKD) have not been thoroughly investigated. METHODS:PENK serum levels were cross-sectionally related to renal and cardiometabolic markers across three large cohorts (total N = 4722), including the Leipzig-CKD cohort (N = 581), as well as the population-based LIFE-Adult (N = 3093) and Sorbs (N = 1048) cohort. Furthermore, a longitudinal analysis was performed in the Leipzig-CKD cohort to assess the association between circulating PENK levels at baseline and a composite 4-point major adverse renal events (4P-MAKE) endpoint comprising incident or worsening kidney disease, development of kidney failure (KF) requiring kidney replacement therapy or kidney transplantation, and renal deaths. RESULTS:In the entire cross-sectional cohort (N = 4722), PENK levels were strongly and inversely correlated with markers of renal function, i.e. estimated glomerular filtration rate (eGFR), creatinine and urinary albumin-creatinine ratio. In multivariable analysis, eGFR was the strongest, independent and inverse predictor of circulating PENK levels after adjustment for age and sex, as well as markers of obesity, glucose intolerance, dyslipidemia and inflammation. Multivariable Cox regression analysis revealed that baseline circulating PENK levels were strongly associated with an increased risk of the 4P-MAKE over a median follow-up of 8.7 years. Each doubling of PENK levels was associated with a >2-fold increase in the risk of developing 4P-MAKE, even after adjusting for various clinical and metabolic variables. PENK provided added value for predicting adverse renal outcomes. CONCLUSIONS:PENK levels are strongly, independently, and inversely associated with renal function in cross-sectional analysis. Furthermore, baseline circulating PENK levels are predictive of a 4P-MAKE in patients and added independent, prognostic value for identification of patients at risk for adverse renal outcomes.

We examined whether long-term exposure to visceral-adipose-tissue (VAT) influences brain atrophy and cognitive performance years after lifestyle intervention. In the Follow-Interventions-Trials (FIT) project, 533 adults (age=61.4 y, 86% men) from four prior 18-24-month lifestyle randomized-clinical-trials underwent abdominal/brain magnetic-resonance-imaging (MRI)s and Montreal-Cognitive-Assessment (MoCA) testing 5-16 y after interventions. Lower VAT exposure, calculated by area-under-the-curve, from baseline, post-intervention, and follow-up, independently resulted in higher MoCA scores. VAT loss during intervention predicted higher brain volumes at follow-up, independent of weight loss. Among participants with three brain and VAT MRI scans, lower long-term VAT was associated with a slower rate of brain atrophy. These patterns were not observed for deep/superficial subcutaneous-adipose-tissues. Improved glycemic control parameters, rather than lipid or inflammatory markers, were mostly related to the favorable longitudinal brain outcomes. This long-term, large-scale intervention and follow-up MRI study suggests that sustained visceral fat loss, rather than weight loss, is linked to better cognition and attenuation of brain atrophy years later, mainly via improved glycemic control. Trial registration: DIRECT (Clinical-trials-identifier: NCT00160108); CASCADE (Clinical-trials-identifier: NCT00784433); CENTRAL (Clinical-trials-identifier: NCT01530724); DIRECT-PLUS (Clinical-trials-identifier: NCT03020186).

BACKGROUND:During the 18-month CENTRAL (Effect of Weight-Loss Diet Strategies and Exercise on Dynamics of Body Fat Depots and Metabolic Rate) and DIRECT-PLUS (Dietary Intervention Randomized Controlled Trial Polyphenols Unprocessed) randomized controlled trials, participants achieved considerable reductions in abdominal and ectopic fat. We examined the long-term postintervention cardiometabolic profile associated with these changes. METHODS:We invited participants from CENTRAL (2012-2014) and DIRECT-PLUS (2017-2018), which evaluated dietary patterns (low-fat, healthy dietary guidelines and Mediterranean diet variants, including standard, low-carbohydrate, and polyphenol-enriched "green" Mediterranean diets) combined with structured physical activity. Participants underwent additional magnetic resonance imaging of visceral adipose tissue, deep subcutaneous adipose tissue (SAT), superficial SAT, intrahepatic fat, and intrapancreatic fat, along with clinical follow-up measurements, 5 and 10 years after completion of the trials. RESULTS:We reached 366 out of 381 eligible participants (96%) for follow-up. Despite complete weight regain, waist circumference and abdominal fat depots, including visceral adipose tissue, deep SAT, and superficial SAT, partially preserved their intervention-induced achievements at long-term follow-up (false discovery rate ≤0.01 for all). In contrast, postintervention reductions of intrahepatic fat and intrapancreatic fat were fully and excessively gained during follow-up, respectively (false discovery rate ≤0.01 for both). Each 10% intervention-induced loss of visceral adipose tissue, superficial SAT, and intrapancreatic fat were associated with long-term postintervention improvements in Metabolic Score for Insulin Resistance, composite risk score, and Metabolic Syndrome Severity Score (meta-analysis models adjusted to weight change, Mediterranean diet adherence and physical activity scores at follow-up, and further measures; all P<0.05). Only 10% visceral adipose tissue loss, however, was independently associated with a 28% lower risk of incident type 2 diabetes (hazard ratio, 0.72 [95% CI, 0.54-0.94]; multivariable model) during follow-up. CONCLUSIONS:This 5- and 10-year follow-up of 18-month clinical trials suggests that diet and physical activity lifestyle interventions may yield long-term improvements in cardiometabolic measures despite weight regain. A 10% reduction in visceral fat due to lifestyle interventions may reduce future type 2 diabetes risk by nearly 30%. Visceral fat loss rather than weight loss emerges as a key target for durable cardiometabolic health. REGISTRATION:URL: https://www.clinicaltrials.gov; Unique identifiers: NCT01530724 and NCT03020186.

BACKGROUND:& Aims: Folate metabolism can be regulated by diet and genetic variation, particularly Methylenetetrahydrofolate reductase (MTHFR) rs1801133 polymorphism. We explored the effect of plant-based green-Mediterranean (green-MED) diet on serum folate and whether this effect varies by genetic variation in folate metabolism over 18-months METHODS: In the DIRECT-PLUS trial, 294 participants were randomized to healthy dietary guidelines (HDG), MED, or plant-based green-MED diets. Both isocaloric MED groups consumed walnuts (28g/day), while green-MED group also avoided red/processed meat, consumed Wolffia-globosa Mankai green shake (500 ml/day), and green-tea (3-4 cups/day). MRI assessed visceral adipose tissue (VAT). MTHFR rs1801133 single-nucleotide-polymorphism (SNP), and folate-pathway mRNA expression were evaluated. RESULTS:Participants (age = 50.1years; 90.6% men; body-mass-index = 31.4 kg/m2; serum folate = 7.6 ng/mL; rs1801133 frequencies: CC = 38.5%; CT = 49.8%; TT = 11.7%) had 89% 18-month retention rate. Folate deficiency at baseline was rare (∼3%, n = 8), indicating effects within the normal range. After 18-months of intervention, the green-MED diet significantly increased serum folate (+1.2 ng/mL) compared to MED (+0.41 ng/mL) and HDG diets (+0.1 ng/mL), p < 0.05 between groups, with higher Mankai intake strongly associated with folate elevation (p = 0.003). rs1801133 TT-genotype carriers had lower folate at baseline (p = 0.037) and post-intervention (p = 0.04; recessive mode-of-inheritance), while CC/CT-genotype carriers showed the highest folate increase following the green-MED intervention (p = 0.03). TT-genotype carriers showed lower baseline MTHFD2 mRNA expression (p = 0.005), but an enriched transcription, compared to CC/CT-carriers post-intervention (p = 0.001). Mankai intake interacted with rs1801133 genotype on VAT (p = 0.028) and Framingham risk score (p = 0.024), with CC/CT-carriers showing dose-response benefits. CONCLUSIONS:The green-MED diet, enriched with Mankai, significantly increased serum folate compared with the MED and HDG diets. The response differed by MTHFR rs1801133-genotype, with CC/CT carriers showing greater increases in folate. These findings support a gene-diet interaction with potential implications for visceral adiposity and cardiometabolic risk.

Acute SARS-CoV-2 infection has been implicated in the development of endothelial dysfunction and metabolic alterations. These disturbances may contribute to the pathophysiology of post-COVID-19 syndrome (PCS), a multifaceted condition characterized by persistent symptoms, including neuropsychiatric symptoms. The diagnosis of PCS primarily relies on symptom-based criteria. Here, we aimed to identify biomarkers associated with PCS and disease severity. This prospective single-center cohort study investigated soluble blood biomarkers related to endothelial dysfunction and amino acid, fatty acid, carnitine, eicosanoid and resolvin metabolism in individuals post-acute SARS-CoV-2 infection with or without PCS compared with individuals without documented SARS-CoV-2 infection. Additionally, we explored the association between these biomarkers and PCS-related fatigue severity as assessed by the Multidimensional Fatigue Inventory (MFI). At a median of 37.4 weeks after SARS-CoV-2 infection, participants with prior infection showed higher levels of soluble thrombomodulin (TM) and L-lactate dehydrogenase (LDH) than those without previous infection. Alterations in arginine biosynthesis and taurine and hypotaurine metabolism indicate disruption of the NO-metabolism. These findings were made in participants without and with symptoms of PCS. In participants with PCS-related high fatigue severity, concentrations of the polyunsaturated fatty acid (PUFA) linoleic acid (LA), and the monounsaturated fatty acids (MUFAs) oleic acid (OA) and palmitoleic acid (PA) were higher than in participants with low fatigue severity. Alterations in markers of endothelial dysfunction and NO-metabolism are detectable at a median of 37.4 weeks after SARS-CoV-2 infection independent of PCS-related fatigue severity. Additionally, in individuals with high PCS-related fatigue severity, specific fatty acid alterations were observed.

Macrophage efferocytosis, essential for the resolution of inflammation and plaque stability in atherosclerosis, is impaired in diabetes. Thrombomodulin (TM) and endothelial protein C receptor (EPCR), key mediators of protein C activation (PC), have vasculoprotective and anti-inflammatory roles, yet their involvement in macrophage efferocytosis in diabetes-induced atherosclerosis remains unclear. Here, we demonstrate that expression of EPCR was reduced in atherosclerotic lesions of diabetic patients compared to non-diabetic controls. In parallel, efferocytosis was impaired in atherosclerotic lesions and in monocytes derived macrophages of diabetic patients. In vitro, treatment with activated PC (aPC) or its cytoprotective selective variant (3K3A-aPC) restored high glucose-impaired macrophage efferocytosis. Mechanistic studies revealed that aPC restored efferocytosis through Arginase-1 and modulation of Rac1-ATF6 signaling. Additionally, macrophage protease-activated receptor 1 (PAR1) was identified as the key receptor mediating aPC's effects on efferocytosis. Mimicking biased PAR-1 signaling via parmodulin-2 reverses glucose impaired efferocytosis. In vivo, aPC treatment of diabetic ApoE-/- mice increased MerTK expression in atherosclerotic lesions. aPC's vasculoprotective effects, including the reduction of plaque size, were abrogated upon MerTK inhibition using morpholinos, underscoring the pivotal role of MerTK in mediating aPC's atheroprotective actions. These findings suggest that impaired TM-PAR1-aPC signaling contributes to defective macrophage efferocytosis in diabetes-associated atherosclerosis and that aPC-based therapies may offer a novel strategy to enhance macrophage function and prevent diabetes induced atherosclerosis.

Obesity is recognized as a pro-thrombotic condition, yet the extent of coagulation activation across biomarkers remains unclear. This meta-analysis evaluates the impact of obesity on parameters-D-dimer, fibrinogen, plasminogen activator inhibitor-1 (PAI-1), von Willebrand factor (vWF), factor VIII (FVIII), and endogenous thrombin potential (ETP)-in children and adults. METHODS:Sixty-four studies comprising 59,503 individuals were analyzed. Plasma biomarker levels were compared between non-obese and obese groups using standardized mean differences (SMDs), with subgroup analyses. RESULTS:D-dimer was significantly elevated in adults with obesity (SMD 1.36, 95% CI 0.47-2.25, p = 0.003) and children with obesity (SMD 0.77, 95% CI 0.19-1.36, p = 0.009), indicating increased fibrin turnover. Fibrinogen levels were markedly higher in both adults (SMD 1.17, 95% CI 0.14-2.20, p = 0.03) and children (SMD 1.43, 95% CI 0.93-1.92, p < 0.0001). PAI-1 showed the most pronounced increase in adults (SMD 2.30, 95% CI 0.1.51-3.09, p < 0.0001) and children (SMD 3.54, 95% CI 1.65-5.43, p = 0.0002). FVIII levels were modestly elevated (SMD 0.52, 95% CI 0.10-0.94, p = 0.02), whereas vWF levels showed inconsistent changes. ETP was significantly higher in obesity, in children (SMD 1.06, 95% CI 0.24-1.88, p = 0.01) and adults (SMD 0.71, 95% CI 0.46-0.97, p < 0.0001). Gender-stratified data indicated higher PAI-1, fibrinogen, and ETP levels in females. CONCLUSION:Obesity is associated with increased coagulation activation, suggesting a pro-thrombotic shift. These findings support the need for age- and gender-specific research into obesity-related hemostatic alterations.

Introduction:Kallikrein-related peptidase 7 (KLK7) is a protease implicated in metabolic disease and obesity. Patients with chronic kidney disease (CKD) exhibit several cardio-metabolic comorbidities and increased mortality. The goal of this study was to investigate the associations of KLK7 levels with renal function and clinical outcomes in patients with CKD. Methods:Baseline KLK7 serum levels were cross-sectionally related to renal and cardiometabolic markers in the Leipzig-CKD cohort (n=542). Longitudinal Cox Regression analyses (n=472) were performed to associate baseline circulating KLK7 concentrations and risk of major adverse renal (MARE) and cardiovascular (MACE) events, as well as all-cause mortality. Additionally, mRNA expression of Klk7 and related genes was examined in CKD versus control mice using bulk RNA sequencing. Results:KLK7 levels were inversely associated with markers of renal function, i.e. estimated glomerular filtration rate (eGFR), and inflammation, i.e. C-reactive protein, in multivariable regression. Longitudinal analyses associated higher baseline KLK7 with lower all-cause (adjusted HR [95% CI]: 0.65 [0.49-0.86], p=0.003) and non-cardiovascular (adjusted HR [95% CI]: 0.56 [0.40-0.78], p=0.001) mortality over a median follow-up of 7.6 years. A KLK7 threshold of 1383.6 pg/ml stratified patients into low- and high-risk groups for mortality. No associations were found for MARE or MACE. In tissues of CKD and control mice, no significant changes in Klk7 mRNA expression were found. Discussion:Circulating KLK7 associates inversely with renal function and inflammation, likely reflecting reduced renal clearance. Higher circulating KLK7 independently predicts lower all-cause mortality, whereas baseline KLK7 was not related to composite renal and CV outcomes.

OBJECTIVES:The adrenal glands are the main source of 11-oxygenated androgens (11-OAs), including the potent 11-ketotestosterone (11-KT) and the weaker metabolites 11β-hydroxyandrostenedione (11-OHA4), 11β-hydroxytestosterone (11-OHT) and 11-ketoandrostenedione (11-KA4). Due to their adrenal origin, 11-OAs allow the differentiation of adrenal and extra-adrenal-produced androgens. However, their clinical use is limited due to the lack of robust age- and sex-specific reference intervals (RIs). METHODS:To establish RIs from infancy throughout childhood and adulthood, we performed simultaneous quantification of 11-OHA4, 11-OHT, 11-KA4, 11-KT, and clinically relevant steroid hormones via LC-MS/MS. We analyzed 3,796 serum samples from 2,505 healthy individuals, aged 0.25-80 years, encompassing minipuberty and pubertal stages. RESULTS:11-OA serum levels increase from infancy throughout childhood and adulthood, with the most pronounced increase during puberty. Significant sex differences were observed, and age-, sex-, and puberty-dependent RIs were established. In contrast to testosterone, 11-OAs exhibited a comparable pattern during the first year of life in both sexes. At 3 and 6 months of age - when testosterone levels were 100- and 10-fold higher, respectively, in males than in females - no correlation was observed between testosterone and the adrenal-derived androgens (11-OAs, A4, and DHEAS) in males. CONCLUSIONS:Established age-, sex-, and puberty-dependent RIs support the clinical interpretation of 11-OA measurements in disorders characterized by androgen excess. The differences in the regulation of classical androgens and 11-OAs during minipuberty suggest that the transient increase in androgens during the first months of life is most likely due to activation of the hypothalamic-pituitary-gonadal axis.

Preeclampsia is a pregnancy complication associated with de novo gestational hypertension, proteinuria, and dysregulations in the coagulation and fibrinolytic systems. Platelet-derived factors include various growth and differentiation factors, such as transforming growth factor (TGF)-β. The TGF-β type III receptor (TGFBR3), also known as betaglycan, is a co-receptor for the TGF-β superfamily and an important regulator of reproduction and fetal development. Here, we test the hypothesis whether or not TGFBR3 is involved in platelet-mediated upregulation of placental plasminogen activator inhibitor-1 (PAI-1). Perivillous fibrin deposits and PAI-1 levels were analyzed via immunohistochemistry, gene expression as well as protein expression in healthy and preeclamptic placental tissue. Placental villi and trophoblast cell line BeWo were incubated in presence or absence of washed platelets from pregnant women, platelet-derived soluble factors, or platelet-derived extracellular vesicles. Additionally, BeWo cells were treated with TGF-β-receptor inhibitors and silenced for TGFBR3 expression. PAI-1 and its encoding gene SERPINE1, as well as perivillous fibrin deposits, were significantly increased towards term of gestation and showed a significant upregulation in placental villi from preeclamptic pregnancies. Furthermore, SERPINE1 and PAI-1 were substantially upregulated in response to washed platelets and platelet-derived factors, but not to platelet-derived extracellular vesicles. The upregulation of SERPINE1 was blocked to control levels with the TGF-β inhibitor P144, whereas silencing of TGFBR3 had no effect. Our results suggest that platelet-derived factors induce placental PAI-1 expression independently of TGF-β receptor 3. Increased maternal platelet activation may contribute to significantly upregulated placental PAI-1 and perivillous fibrin deposits in preeclampsia.

ABSTRACT Aims Lifestyle‐induced weight loss (LIWL) is considered an effective therapy for the treatment of metabolic syndrome (MetS). The role of differentially expressed genes (DEGs) in adipose tissue function and in the success of LIWL in MetS is still unclear. We investigated the effect of 6 months of LIWL on transcriptional regulation in subcutaneous adipose tissue (SAT). Aiming to identify a LIWL‐associated “gene signature” in SAT, DEGs were fitted into a linear regression model. Materials and Methods The study is embedded in a prospective, two‐arm, controlled, monocentric, randomized, 6‐month interventional trial in individuals with MetS following LIWL. The trial included 43 nonsmoking, nondiabetic men aged 45–55 years with MetS. Results In total, we identified 642 DEGs in SAT after 6 months of LIWL. The identified DEGs were validated in two cross‐sectional cohorts analyzing SAT from individuals with and without obesity. Gene enrichment analysis of the DEGs revealed the strongest association with cholesterol metabolic processes. Accordingly, DEGs were correlated with the lipid parameters HDL cholesterol, LDL cholesterol, and triglycerides in corresponding serum samples. We identified 3 genes with an AUC of 0.963 (95% CI: 0.906–1.0) associated with a loss of more than 10% of initial body weight that was maintained for at least 12 months after LIWL, namely SUMO3 (Small ubiquitin‐related modifier 3), PRKG2 (Protein Kinase CGMP‐Dependent 2), and ADAP2 (ArfGAP with Dual PH Domains 2). Conclusion In summary, we have identified DEGs in SAT after LIWL, which may play an important role in metabolic functions. In particular, altered gene expression in SAT may predict sustained weight loss.

Background: Preeclampsia is a vascular complication of pregnancy with limited therapeutic options. It is associated with hypertension and an increase in angiogenic factor soluble fms-like tyrosine kinase-1 (sFlt-1)/placental growth factor. Based on its onset, preclampsia can be categorized into early-onset (E-PE) or late-onset (L-PE) preeclampsia. Thrombo-inflammation, hallmarked by maternal platelet activation and sterile inflammation, is associated with pathophysiology of preeclampsia. However, whether these mechanisms are differentially regulated in E-PE vs L-PE remains unknown. Objectives: We aim to study the role of maternal platelet activation, inflammation and endothelial dysfunction in E-PE vs L-PE. Methods: Flow-cytometry analysis of platelet activation (P-selectin and active αIIbβ3) was conducted in whole blood from pregnant women with E-PE, L-PE and gestational age-matched patients. Plasma was evaluated for interleukin (IL)-1β and soluble vascular cell adhesion molecule 1 (sVCAM-1). Results: An increase in P-selectin and active αIIbβ3 expressing platelets in both forms of preeclampsia (n = 22) was observed compared with their gestational age-matched controls (n = 18). Similarly, an increase in plasma IL-1β and sVCAM-1 was observed in both forms of preeclampsia, suggesting inflammation and endothelial dysfunction, respectively. Maternal platelet activation (P-selectin positive platelets) was linked with disease severity (sFlt-1/placental growth factor) and maternal plasma IL-1β and sVCAM-1 only in late-onset preeclampsia. A statistically significant correlation with αIIbβ3 expressing platelets and sFlt-1, IL-1β, and sVCAM-1 was not observed. Conclusions: These findings identify that thrombo-inflammation is regulated in L-PE and E-PE through likely disjunct mechanisms supporting a role of maternal factors (eg, maternal platelet activation) involved in L-PE. Further studies with a larger cohort of patients are required to fully elucidate the mechanistic relevance of these findings.

AIMS:We explored the manifestations of individual weight loss (WL) response to long-term lifestyle interventions on cardiometabolic risk. METHODS AND RESULTS:We pooled data from three large long-term lifestyle WL-intervention trials: 24-month DIRECT (ClinicalTrials.gov: NCT00160108; n = 322; 87% adherence), 18-month CENTRAL (ClinicalTrials.gov: NCT01530724; n = 278; 86% adherence), and 18-month DIRECT PLUS (ClinicalTrials.gov: NCT03020186; n = 294; 89% adherence). We analyzed longitudinal changes in cardiometabolic risk markers, including anthropometrics, blood biomarkers, and magnetic-resonance-imaging-assessed fat depots, and measured DNA-methylation, proteomics, and metabolomics. Among trial completers (n = 761, mean age = 50.4 years; 89% men, baseline body-mass-index = 30.1 kg/m2), mean WL was -3.3 kg (-3.5%). We classified participants as Successful-WL (36%) with relative-WL > 5%, WL-Resistant (28%) who did not lose or gained weight, and Moderate-WL (36%) with WL between 0% and 5%. Successful-WL achieved the greatest improvements in multiple health indicators. However, the WL-Resistant also showed some significant improvements, with increased high-density-lipoprotein-cholesterol (HDLc) and decreased leptin and visceral fat (P < 0.05 vs. baseline). Overall, each 1 kg sustained lifestyle-induced WL was associated with improvements in lipid markers and insulin resistance [HDLc (+1.44%), triglycerides (-1.37%), insulin (-2.46%), HOMA-IR (-2.71%), leptin (-2.79%)] and intrahepatic-fat regression (-0.49 absolute-units)] and modest but significant change in systolic and diastolic blood pressures (-0.26% and -0.36%). We identified 12 significant methylation sites that are associated with Successful-WL (FDR < 0.05; AUC = 0.73). CONCLUSION:While only ∼one-third of individuals achieved long-term WL, the Moderate-WL and WL-Resistant individuals could benefit improvements in visceral adiposity and cardiometabolic risk by shifting towards a healthy lifestyle pattern, beyond WL. Site-specific DNA methylation may predict an individual's likelihood of successful WL. REGISTRATION:NCT00160108, NCT01530724, NCT03020186.

BACKGROUND:Cardiogenic shock (CS) caused by acute myocardial infarction (AMI) is a critical condition with high mortality rate. Insulin-like growth factor binding protein 2 (IGFBP-2) is dysregulated in cardiovascular diseases. The purpose of the present study was to investigate the prognostic value of IGFBP-2 in patients with AMI-CS. METHODS:This study is a post-hoc analysis of the randomized multicentre CULPRIT-SHOCK trial. IGFBP-2 levels were measured in serum samples from 423 patients using commercially available enzyme-linked immunosorbent assay (ELISA) kits. Associations of IGFBP-2 with 30-day and one-year mortality were investigated. RESULTS:Median IGFBP-2 concentration was 415 ng/ml (IQR 274-699 ng/ml). Patients with IGFBP-2 ≥ median demonstrated higher 30-day (54 % vs. 37 %; p < 0.001) and one-year mortality (60 % vs. 42 %; p < 0.001) compared to the < median group. Higher IGFBP-2 concentrations were associated with increased 30-day and one-year mortality, irrespective of it being analysed as continuous or categorical variable (per 100 ng/ml IGFBP-2, hazard ratio (HR) 1.06; 95 % confidence interval (CI) 1.04-1.09; p < 0.001, respectively; IGFBP-2 ≥ vs. < median, HR 1.70, 95 % CI 1.23-2.35, p = 0.001 and HR 1.72, 95 %CI 1.27-2.33, p < 0.001). Furthermore, IGFBP-2 ≥ median was associated with increased 30-day (HR 1.70; 95 %CI 1.23-2.35; p = 0.001) and one-year mortality (HR 1.72; 95 %CI 1.27-2.33; p < 0.001), even after adjustment for established prognostic factors. CONCLUSIONS:In AMI-CS, elevated levels of IGFBP-2 were associated with higher mortality at 30 days and one year after admission. IGFBP-2 represents a promising prognostic biomarker and could add value to risk stratification in this high-risk patient cohort, potentially informing early clinical decision-making.

BACKGROUND AND AIMS:We explored whether changes in serum proteomic profiles differed between participants with distinct brain aging trajectories, and whether these changes were influenced by dietary intervention. METHODS:In this secondary analysis of the 18-month DIRECT PLUS trial, 294 participants were randomized to one of three arms: 1) Healthy dietary guidelines (HDG); 2) Mediterranean (MED) diet (+440 mg/day polyphenols from walnuts); or 3) low red/processed meat green-MED diet (+1240 mg/day polyphenols from walnuts, Mankai plant, and green tea). We measured 87 serum proteins (Olink-CVDII). We used Magnetic-Resonance-Imaging (MRI)-assessed 3D-T1-weighted brain scans for brain age calculation (by convolutional neural network) to identify protein markers reflecting the brain age gap (BAG; deviation of MRI-assessed brain age from chronological age). RESULTS:At baseline, lower weight, waist circumference, diastolic blood pressure, and HbA1c parameters were associated with a younger brain age than expected. Specifically, higher levels of two proteins, Galectin-9 (Gal-9) and Decorin (DCN), were associated with accelerated brain aging (larger BAG). A proteomics principal component analysis (PCA) revealed a difference in PC1 between the two time-points for participants with accelerated brain aging. Between baseline and 18 months, Gal-9 significantly decreased among individuals who completed the intervention with attenuated brain aging, while DCN significantly increased among those who completed the trial with accelerated brain aging. A significant interaction was observed between the green-MED diet and proteomics PCA, resulting in a beneficial change compared to the HDG. Participants in the green-MED diet significantly decreased Gal-9 compared to the HDG diet and from baseline. CONCLUSIONS:Higher serum levels of Gal-9 and DCN may indicate an acceleration of brain aging and could be reduced by a green-MED/high-polyphenol (green tea and Mankai) and low-red/processed meat diet. TRIAL REGISTRATION NUMBER:NCT03020186.