Early-onset obesity, especially in girls, is frequently associated with advanced puberty; a phenomenon bound to increased risk of long-term complications. Hypothalamic endoplasmic reticulum (ER) stress has been implicated in the pathophysiology of obesity and its comorbidities. However, its contribution to pubertal disorders associated with obesity remains unexplored. We report herein that central ER stress drives obesity-induced precocious female puberty. Hypothalamic expression of key ER stress markers was blunted during normal pubertal transition and altered in female rats with early-onset obesity and advanced puberty, which displayed increased levels of p-PERK and p-eIF2α, and reduced ATF6α content. Central stimulation of hypothalamic ER stress with Thapsigargin in prepuberal lean female rats mimicked advanced puberty onset caused by obesity, without changes in body weight. This phenomenon seemingly involves a circuit including the hypothalamic arcuate nucleus (ARC), since obesity-induced precocious puberty was largely prevented by alleviation of ER stress via virogenetic overexpression of the ER chaperone, GRP78, in the ARC, but not in the paraventricular nucleus, in female rats. In addition, expression analyses of ER stress markers in mouse Kiss1 neurons isolated from juvenile and pubertal female mice revealed increased expression of Perk and Ire1α mRNAs in Kiss1ARC neurons in early-overfed mice at the juvenile stage, while Xbp1s/u expression ratio was significantly increased during juvenile-pubertal transition in overweighed mice. Collectively, our data uncover a relevant role of hypothalamic ER stress in the control of female puberty and the pathogenesis of obesity-induced pubertal alterations.
Background Obesity-related cardiometabolic disease is linked to impaired adipose tissue function, but the underlying molecular programs are difficult to assign to specific adipose-resident cell types, to mechanistically connect to the activation state of macrophages, and to distinguish from alterations that may normalize with weight loss. Methods We integrated a layered design combining untargeted proteomics and lipidomics to define obesity-associated, cell-type-resolved molecular phenotypes across adipocytes and adipose microvascular endothelial cells, explore whether an inflammatory milieu reproduces adipose-resident cell dysfunction, and identify features that show evidence of recovery after weight loss. Next, we validated in adipose tissue transcriptomes the elements that exhibit the strongest association with dyslipidaemia, hypertriglyceridemia, and/or hyperglycaemia to identify gene signatures of cardiometabolic relevance. Results Adipocytes from subjects with obesity show suppression of mitochondrial energy metabolism together with impaired lipid plasticity, as reflected by triglyceride remodelling. By mimicking an inflammatory milieu with macrophage-conditioned media, we reproduced many of these changes in adipocyte cultures. Endothelial cells exhibited yet another, opposite trajectory in obesity, with reduced cell-cycle signalling and increased mitochondrial activation, which were recapitulated in vitro when these cells were exposed, respectively, to the secretions of inflamed macrophages and adipocytes. Bulk adipose tissue proteomes and lipidomes showed evidence of metabolic improvement after weight loss, including restoration of mitochondrial and substrate-handling pathways alongside reciprocal triglyceride remodelling. Together with inflammation-responsive adipocyte mitochondrial and lipid-handling dysfunction, our cell-type-informed framework probes macrophage and adipocyte-to-endothelial activation in obesity, and delineates cross-context cellular programs associated with weight loss. Notably, when integrated with transcriptomic resources, these layers of information prioritized determinants linked to impaired metabolism, and were used to generate models that can assess cardiometabolic vulnerability in subjects with obesity (AUROC values between 0.88 and 0.99). Conclusions Our study reveals adipocyte and endothelial cell-specific elements acting as molecular signatures of adipose tissue inflammation with cardiometabolic implications.
Spermidine administration ameliorates hepatic steatosis and cardiometabolic dysfunction in animal models. However, evidence in humans remains limited. We aimed to explore the 1-year longitudinal associations between changes in dietary spermidine intake and changes in hepatic function indexes and cardiometabolic traits in overweight and obese older adults with metabolic syndrome. We used baseline and 1-year follow-up data from 2664 participants from the PREDIMED-Plus trial. Dietary spermidine intake was estimated using a semi-quantitative food frequency questionnaire. Time series clustering identified temporal patterns of 1-year change in spermidine intake. Linear mixed-effects models assessed the associations between spermidine intake clusters and changes in hepatic and cardiometabolic markers. Three distinct spermidine intake patterns were identified across baseline, six months and 1 year follow-up. Cluster 3, characterized by the highest baseline and increased 1-year spermidine intake, was associated with mean reductions in fatty liver index (− 8.97 [− 9.96 to − 7.97], p-int < 0.001), hepatic steatosis index (− 1.88 [− 2.13 to − 1.63], p-int < 0.001), alanine aminotransferase (− 2.93 [− 3.83 to − 2.02] U/L, p-int < 0.05), aspartate aminotransferase (− 1.11 [− 1.76 to − 0.45] U/L, p-int < 0.05) and glycated hemoglobin levels (− 0.14 [− 0.18 to − 0.10]
ABSTRACT Detection of GES-type carbapenemases remains challenging because of their low prevalence and frequently weak hydrolytic activity against carbapenems. Carbapenem inactivation method (CIM)-based assays are widely used as phenotypic screening tools for carbapenemase detection; however, their performance in large collections of GES producers has not been systematically evaluated. We assessed the performance of CIM, modified CIM (mCIM), and CIM-Tris in a diverse collection of GES-producing clinical isolates, including 110 Enterobacterales and 108 Pseudomonas aeruginosa, recovered from Spanish hospitals (2010–2024), and 10 Acinetobacter baumannii isolates, mostly obtained from a hospital in Egypt. Whole-genome sequencing was carried out for species confirmation and resistome analysis. Meropenem MICs were determined by broth microdilution. Overall, 92.1% of isolates were GES-carbapenemase producers (CP), whereas 7.9% expressed GES-type extended-spectrum β-lactamases (ESBLs). In Enterobacterales (predominantly carrying bla GES-6 ), mCIM improved sensitivity compared with CIM (63.6% vs 40.0%), although many isolates remained undetected due to low meropenem MICs (MIC 50 , 0.5 µg/mL). In CP- P. aeruginosa (mainly bla GES-5 ), CIM, mCIM, and CIM-Tris showed sensitivities of 89.1%, 94.6%, and 100%, respectively; however, CIM-Tris yielded false-positive results in 50% of non-CP isolates (mostly bla GES-1 producers). Meropenem MICs in P. aeruginosa were higher (MIC 50 , >32 µg/mL). In A. baumannii , CIM-Tris improved sensitivity compared with CIM (100% vs 25.0%). These findings indicate that CIM-based methods can detect GES-type carbapenemases, but performance varies according to bacterial species and GES variant, and reduced specificity may occur in isolates producing GES-type ESBLs. Complementary molecular testing may therefore be necessary to ensure accurate detection of GES-type carbapenemases in routine clinical laboratories. IMPORTANCE GES-type carbapenemases represent an important but underrecognized diagnostic challenge due to their low global prevalence, heterogeneous hydrolytic activity, and the limited performance data available for routine phenotypic detection methods. Although CIM-based assays are widely implemented in clinical microbiology laboratories for carbapenemase screening, their performance against GES-producing organisms has not been comprehensively evaluated across different bacterial genera and GES variants. In this study, we evaluated the performance of CIM, modified CIM (mCIM), and CIM-Tris in a large multicenter collection of well-characterized GES-producing clinical isolates, including Enterobacterales, Pseudomonas aeruginosa , and Acinetobacter baumannii . Our findings demonstrate substantial variability in assay performance according to bacterial species and GES variant. Notably, mCIM improved sensitivity among Enterobacterales with low meropenem MICs, whereas CIM-Tris achieved excellent sensitivity in P. aeruginosa and A. baumannii but at the expense of reduced specificity in isolates producing GES-type ESBLs. To the best of our knowledge, this is the first study directly comparing multiple CIM-based approaches in such a large and taxonomically diverse collection of GES-producing isolates.
Dual-specificity tyrosine-phosphorylation-regulated kinase 2 (DYRK2) is emerging as a pivotal player in cancer biology due to its pleiotropic cellular roles, functioning either as a tumor suppressor or an oncogene in a tissue- and genotype-dependent manner. DYRK2 regulates critical processes including cell cycle progression, DNA damage response, and stress signaling. As a tumor suppressor, DYRK2 facilitates the degradation of oncogenic proteins (e.g., c-Myc, SNAIL, TERT) and enhances apoptosis via p53 activation. Loss of DYRK2 has been associated with poor prognosis and chemoresistance in cancers such as colorectal, breast, and liver. Conversely, DYRK2 overexpression in certain tumors, including triple-negative breast cancer and ovarian carcinoma, supports oncogenic roles by enhancing proteasome function, promoting epithelial-mesenchymal transition, and contributing to therapy resistance. This functional dichotomy is modulated by post-translational modifications, microenvironmental stress, epigenetic regulation, subcellular localization, and interaction with protein complexes, underscoring the complexity of DYRK2 regulation. Therapeutically, DYRK2 represents a promising but context-sensitive target, with inhibitors showing efficacy in preclinical models of proteasome-addicted cancers, although precise biomarkers and cancer subtype stratification remain critical. Future directions include refining in vivo models and delineating the role of DYRK2 in cancer stem cell dynamics and immune modulation, emphasizing that understanding its context-dependent activity is essential to exploit its full therapeutic potential in oncology.
