Mast cells play a crucial role in the pathogenesis of asthma, by releasing inflammatory mediators including mast cell-specific proteases such as tryptase and chymase. However, the exact role of these proteases in asthma is not fully understood. We showed previously that chymase imposes multiple effects on primary human lung fibroblasts (HLFs), and in the present report we addressed the underlying mechanisms. The effects of chymase on HLFs were found to be independent of protease-activated receptors (PARs), as judged by employing PAR agonists and antagonists, despite PAR1 and PAR3 being highly expressed. Further, Western blot analysis revealed that chymase degraded fibronectin and affected the levels and phosphorylation status of multiple signalling factors related to fibronectin, including integrin αVb3, focal adhesion kinase, Src and Akt. This was associated with a decreased motility of chymase-treated HLFs, as assessed by a cell migration assay. Chymase treatment also suppressed the levels and/or the phosphorylation status of CREB and c-Jun. Intriguingly, chymase was shown to induce the degradation of HSP27, an intracellular protein, suggesting that chymase may exert proteolytic activity in the cell interior. In agreement with this, confocal microscopy analysis revealed that chymase was taken up by the HLFs. By using Seahorse technology, we also demonstrate that chymase has the capacity to suppress the metabolic activity of HLFs, without causing cell death. Altogether, these findings reveal a novel role for mast cell chymase in affecting signalling pathways in HLFs, thereby providing insight into how mast cells potentially can affect the lung microenvironment in asthma.

BACKGROUND:Evidence for the role of health behaviour and lifestyle treatment (HBLT) on the effectiveness of glucagon-like peptide-1 receptor agonists (GLP-1RAs) in paediatric obesity is limited. OBJECTIVES:To assess the role of HBLT on the effectiveness of GLP-1RAs in youth with obesity. METHODS:Patients aged 8-18 years with obesity (n = 51) receiving liraglutide were retrospectively divided into continuers (ongoing liraglutide treatment), discontinuers, or switches to semaglutide or metabolic-bariatric surgery. All were offered multidisciplinary HBLT and consecutively allocated to either high-frequency (HF) HBLT (≥ 26 contact hours/year) or low-frequency obesity medication (OM)-specific HBLT (9 contacts of 0.5 h/year). RESULTS:After 9.7 ± 6.6 months, a BMI reduction of ≥ 5% and ≥ 10% was observed in 37.3% and 13.7%, respectively. Relative change in BMI was higher in continuers compared to discontinuers (-6.7% ± 7.5% vs. -0.7% ± 5.1%, p = 0.03). Whereas 39.2% of patients continued liraglutide, 33.3% discontinued, mainly due to gastrointestinal symptoms. 27.4% of patients switched treatment to semaglutide or underwent surgery. Patients with combined HBLT and liraglutide had markedly higher odds of continuation (OR 18.5, 95% CI 2.0-929.8, p < 0.01) and greater reductions in BMI, which was by trend higher with OM-HBLT compared to HF-HBLT (BMI change -6.9% ± 7.2% vs. -4.0% ± 6.6%). CONCLUSIONS:GLP-1RAs were effective in the real-world treatment of paediatric obesity. HBLT appears to be essential to increase persistence and efficacy of GLP-1RAs.

INTRODUCTION:We aimed to study the effects of the glucagon-like peptide-1 receptor agonist exenatide on behavioral outcomes in children and adolescents with obesity. METHODS:This study reports pre-specified secondary outcomes of a randomized, double-blind, placebo-controlled trial in adolescents with obesity, aged 10-18 years. Participants (n = 44) were randomized to 6 months treatment with exenatide or placebo, along with a lifestyle intervention consisting of nutritional advice from a dietitian and sessions with a psychologist to optimize physical activity. Change in choices of food and drink, portion sizes, meal frequency, sleep, screen time, and physical activity were assessed by 6-min walking test, accelerometry, and questionnaires. RESULTS:Exenatide treatment was associated with improved adherence to choices of recommended food and drinks (score change 4.34, 95% CI: 1.05-7.63, p = 0.01, Hedges' g 0.82, 95% CI: 0.17-1.49), decreased portion size (score change -4.05, 95% CI: -6.51 to -1.58, p = 0.002, Hedges' g -1.29, 95% CI: -2.13 to -0.45), and increased self-reported physical activity (1.82 h, 95% CI: 0.68-2.96, p = 0.002 Hedges' g 0.97, 95% CI: 0.33-1.62), while no differences in meal frequency, snacking, sleep, screen time, objectively measured physical fitness or activity were found. CONCLUSION:Exenatide treatment was associated with modest improvements in selected self-reported dietary behaviors, including improved adherence to recommended food and drink choices and reduced portion sizes, as well as increased self-reported physical activity, although no changes were observed in objective measures of physical activity or other lifestyle factors.

Glucose uptake in activated CD4+ T cells is essential for increased metabolic needs, synthesis of biomolecules and proliferation. Although, facilitated glucose transport is the predominant route for glucose entry at the time of activation, here we demonstrate role for the sodium-dependent glucose transporter SGLT2. By 72 h after activation, SGLT2 is expressed and functional in the human CD4+ T cells. SGLT2 inhibitors, phlorizin and empagliflozin decreased glucose uptake into the human CD4+ T cells compared to untreated cells. Phlorizin (25 μmol/L) reduced glycolysis at 5.6 mmol/L glucose and IFNγ levels at both 5.6 mmol/L and 16.7 mmol/L glucose. In contrast, empagliflozin (0.5 μmol/L) only decreased IFNγ levels in 16.7 mmol/L glucose. GABA enhanced phlorizin inhibition at both 5.6 mmol/L and 16.7 mmol/L glucose in the presence of insulin. Insulin strengthens GABAA receptors signaling in CD4+ T cells. The results are consistent with expression of SGLT2 after activation of human CD4+ T cells, that facilitates concentrating glucose uptake into the cells, enabling enhanced release of inflammatory molecules like IFNγ. Importantly, inhibition of SGLT2 decreases IFNγ release.

Patients with obesity during childhood have an increased risk of fatal and non-fatal cardiovascular events during adulthood. The severity of obesity is commonly determined by BMI. However, children with relatively low BMI may have high cardiometabolic risk. Indeed, BMI-based obesity classifications might miss children at high cardiometabolic risk. Insulin has been suggested as a marker of cardiometabolic risk. In this study, we therefore estimated and compared cardiometabolic risk using either the BMI standard deviation score (BMI-SDS) or fasting insulin in an international cohort of children and adolescents with obesity and lean controls. Study participants (712 with obesity and 99 lean controls), aged 3 to 18 years, were categorized according to their BMI-SDS as lean or obesity class I, II, or III, or by their fasting insulin quartiles as quartile 1, 2, 3, or 4 with the lean subjects in a separate control group. Prevalence of cardiometabolic risk factors was assessed in each group. Sensitivity and specificity analyses for cardiometabolic risk were conducted for both BMI-SDS and fasting insulin. Multiple regression, logistic regression, and receiver operating characteristic (ROC) analyses were performed between fasting insulin, BMI-SDS and cardiometabolic risk factors. An elevated prevalence of the cardiometabolic risk factors dyslipidemia, dysglycemia and hypertension was observed in both increasing BMI-SDS classes and increasing fasting insulin quartiles. Fasting insulin demonstrated higher areas under the curve (AUC) for detecting dyslipidemia, dysglycemia, and the combination of dyslipidemia, dysglycemia, and hypertension, compared to BMI-SDS. BMI-SDS demonstrated a higher AUC for detecting hypertension compared to fasting insulin. The same patterns were seen for the logistic regression. However, fasting insulin had an overall stronger association with the cardiometabolic risk factors studied compared to BMI-SDS. In children and adolescents with obesity, fasting insulin provides complementary information to BMI-SDS in identifying those with elevated cardiometabolic risk factors. While neither marker alone offers strong predictive accuracy, incorporating fasting insulin into clinical assessment may help prioritize individuals who require more detailed evaluation. An elevated fasting insulin value may warrant further investigation among children and adolescents with obesity, independent of obesity class based on BMI-SDS.

BACKGROUND AND AIMS:Peripheral nerve injuries (PNIs) cause motor and sensory dysfunction. This study assessed cardamonin's effectiveness in enhancing sciatic nerve regeneration and functional recovery in a rat model of nerve crush injury. METHODS:Male rats underwent unilateral sciatic nerve crush and received graded doses of cardamonin (5, 10, 20 mg/kg). The study also included a Sham surgery and a Crush control group. Motor function was assessed with the Sciatic Functional Index (SFI) and sensory function using the Hargreaves test. Electrophysiological measurements, including compound muscle action potential (CMAP) amplitude and nerve conduction velocity (NCV), were recorded at 8 weeks. Histomorphometric analyses of nerve and gastrocnemius muscle were performed, along with quantification of pro- and anti-inflammatory cytokines, neurotrophic factors, and apoptosis markers 7 days post-injury. . RESULTS:Cardamonin treatment significantly improved motor (SFI) and sensory recovery in a dose-dependent manner compared to crush controls (p < 0.001). CMAP and NCV also improved significantly with cardamonin treatment (p < 0.001). Histological evaluation revealed partial restoration of myelin thickness, axon and fiber diameter, and an increase in fiber number (p < 0.01). Cardamonin reduced muscle atrophy and collagen deposition (p < 0.001). Pro-inflammatory cytokines IL-1β, IL-6, and TNF-α were significantly attenuated, while anti-inflammatory cytokines IL-10 and TGF-β, and neurotrophic factors NGF, BDNF, and NT-3 were upregulated dose-dependently (p < 0.05). Caspase-3, -8, and -9 activities were markedly reduced (p < 0.05). CONCLUSION:Cardamonin promotes peripheral nerve regeneration and functional recovery after sciatic nerve crush injury through anti-inflammatory, neurotrophic, and anti-apoptotic effects.

Introduction: Growth charts were not designed to monitor children and adolescents with severe obesity. We evaluate three commonly used international references and their implications for children with severe obesity and develop a BMI growth chart for children with severe obesity, which we call "Reference-point BMI from adjusted World Health Organization (WHO) population" (R-BMI). Method: Growth charts from the WHO, International Obesity Task Force, and CDC were reviewed regarding population, statistical method, and cut-offs. We created the R-BMI chart from the WHO population, with adapted adjustment and reference-point cut-offs, and the layout was updated for better readability. Moreover, an interactive web app was developed for this project at the following link https://child-bmi.serve.scilifelab.se/ with the purpose of visually comparing different BMI references for children with obesity. Results: Three different references for children with severe obesity, with corresponding adjustments, are presented to illustrate implications for researchers and clinicians. Furthermore, R-BMI is presented as a method attempting to address chart challenges related to the extreme BMI. The result is reference curves which share desirable features with established references, while avoiding undesirable curve behavior. Conclusions: Growth charts present challenges for children living with severe obesity, leading to varying approaches and implications of international references. The proposed R-BMI offers monitoring of children with severe obesity that can be used from birth to adulthood. It relates to adult BMI cut-offs and allows for a terminology, and it has a layout with the potential of highlighting changes which may otherwise go unnoticed.

To investigate the prevalence and possible risk factors for the development of impaired glucose metabolism in children and adolescents with obesity. This was a cross-sectional retrospective cohort study, including 634 patients with obesity and 98 normal weight controls aged 4–18 years from the Beta-cell function in Juvenile Diabetes and Obesity (Beta-JUDO) cohort, a dual-centre study at Uppsala University Hospital (Sweden) and Paracelsus Medical University Hospital (Salzburg, Austria) conducted between 2012 and 2021. A longitudinal subgroup analysis, including 188 of these subjects was performed. Impaired glucose metabolism was diagnosed by oral glucose tolerance tests according to American Diabetes Association criteria. The prevalence of impaired glucose metabolism was 72% in Uppsala patients, 24% in Salzburg patients, 30% in Uppsala controls and 13% in Salzburg controls. The prevalence was lower at the follow-up visits compared with baseline both in Uppsala and Salzburg patients. A family history of type 2 diabetes showed the strongest association with impaired glucose metabolism at the follow-up visits besides belonging to the Uppsala cohort. The prevalence of impaired glucose metabolism was extraordinarily high in Swedish children and adolescents with obesity, but decreased during the follow-up period.

BACKGROUND:γ-aminobutyric acid (GABA), known as the main inhibitory neurotransmitter in the brain, exerts immunomodulatory functions by interaction with immune cells, including T cells. Metabolic programs of T cells are closely linked to their effector functions including proliferation, differentiation, and cytokine production. The physiological molecules glucose and insulin may provide environmental cues and guidance, but whether they coordinate to regulate GABA-mediated T cell immunomodulation is still being examined. METHODS:CD4+ T cells that were isolated from blood samples from healthy individuals and from patients with type 1 diabetes (T1D) were activated in vitro. We carried out metabolic assays, multiple proximity extension assay (PEA), ELISA, qPCR, immunoblotting, immunofluorescence staining, flow cytometry analysis, MS-based proteomics, as well as electrophysiology and live-cell Ca2+ imaging. FINDINGS:We demonstrate that GABA-mediated reduction of metabolic activity and the release of inflammatory proteins, including IFNγ and IL-10, were abolished in human CD4+ T cells from healthy individuals and patients with T1D when the glucose concentration was elevated above levels typically observed in healthy people. Insulin increased GABAA receptor-subunit ρ2 expression, enhanced the GABAA receptors-mediated currents and Ca2+ influx. GABA decreased, whereas insulin sustained, hexokinase activity and glycolysis in a glucose concentration-dependent manner. INTERPRETATION:These findings support that metabolic factors, such as glucose and insulin, influence the GABA-mediated immunomodulation of human primary T cells effector functions. FUNDING:The Swedish Children's Diabetes Foundation, The Swedish Diabetes Foundation, The Swedish Research Council 2018-02952, EXODIAB, The Ernfors Foundation, The Thurings Foundation and the Science for Life Laboratory.

AIM:To investigate the prevalence and possible risk factors for the development of impaired glucose metabolism in children and adolescents with obesity.METHODS:This was a cross-sectional retrospective cohort study, including 634 patients with obesity and 98 normal weight controls aged 4-18 years from the Beta-cell function in Juvenile Diabetes and Obesity (Beta-JUDO) cohort, a dual-centre study at Uppsala University Hospital (Sweden) and Paracelsus Medical University Hospital (Salzburg, Austria) conducted between 2012 and 2021. A longitudinal subgroup analysis, including 188 of these subjects was performed. Impaired glucose metabolism was diagnosed by oral glucose tolerance tests according to American Diabetes Association criteria.RESULTS:The prevalence of impaired glucose metabolism was 72% in Uppsala patients, 24% in Salzburg patients, 30% in Uppsala controls and 13% in Salzburg controls. The prevalence was lower at the follow-up visits compared with baseline both in Uppsala and Salzburg patients. A family history of type 2 diabetes showed the strongest association with impaired glucose metabolism at the follow-up visits besides belonging to the Uppsala cohort.CONCLUSION:The prevalence of impaired glucose metabolism was extraordinarily high in Swedish children and adolescents with obesity, but decreased during the follow-up period.

Follicle-stimulating hormone (FSH) is traditionally known for its role in reproduction, but recent studies suggest it may also influence metabolic processes. This study aimed to examine FSH receptor (FSHR) expression in human pancreatic islets and the direct effects of FSH on insulin secretion, as well as explore FSH’s metabolic role during puberty, focusing on enhanced insulin secretion during this critical period. FSHR gene and protein expression were detected in isolated human pancreatic islets and co-localized with insulin-producing beta-cells. Additionally, FSH at prepubertal (0.1 IU/L) and pubertal concentrations (10 IU/L) significantly enhanced glucose-stimulated insulin secretion (GSIS) and increased intracellular cAMP concentrations in intact human pancreatic islets. In children with obesity from the Beta-JUDO cohort (n=608), plasma FSH levels were positively associated with several insulin secretion indices, particularly in pubertal children. These findings suggest that FSH has significant metabolic roles beyond reproduction, involving insulin secretion and potentially contributing to puberty-related hyperinsulinemia and insulin resistance. ### Competing Interest Statement The authors have declared no competing interest.

Introduction: Hudda-Index is a prediction model for fat mass (FM) based on simple anthropometric measures. FM is a crucial factor in the development of comorbidities, i.e., type 2 diabetes. Hence, Hudda-Index is a promising tool to facilitate the identification of children at risk for metabolic comorbidities. It has been validated against deuterium dilution assessments; however, independent validation against the gold standard for body composition analysis, magnetic resonance imaging (MRI), is lacking. The aim of this study was to validate FM calculated by Hudda-Index against FM measured by MRI. The secondary aim was to compare Hudda-Index to other anthropometric measures including body mass index (BMI), BMI-standard deviation score (BMI-SDS), waist/hip-ratio, waist circumference (WC), and skinfold thickness. Methods: The study cohort consists of 115 individuals between the age of 9 and 15 years, recruited at Paracelsus Medical University Hospital in Salzburg (Austria) and Uppsala University Children’s Hospital (Sweden). Anthropometry, blood samples, and oral glucose tolerance tests followed standard procedures. MRI examinations were performed to determine visceral adipose tissue (VAT) and subcutaneous adipose tissue. Results: BMI and WC showed slightly stronger associations with the reference standard VAT (r = 0.72 and 0.70, p < 0.01, respectively) than Hudda-Index (r = 0.67, p < 0.01). There is an almost perfect linear association between BMI and Hudda-Index. Accordingly, BMI and Hudda-Index both showed an acceptable association with cardiometabolic parameters. VAT was strongly associated with markers of liver status (LFF r = 0.59, p < 0.01) and insulin resistance (HOMA-IR r = 0.71, p < 0.01) and predicted metabolic dysfunction-associated steatotic liver disease. Conclusion: BMI, although an imperfect measure, remains the most reliable tool and estimates cardiometabolic risk more reliably than other anthropometry-based measures.

BackgroundFasting levels of glucagon are known to be elevated in youth and adults with type 2 diabetes mellitus (T2D). Children and adolescents with obesity were previously reported to show increasing fasting and post-glucose-challenge hyperglucagonemia across the spectrum of glucose tolerance, while no data are available in those with impaired fasting glucose (IFG).Materials and methodsIndividuals from the Beta-JUDO study population (Uppsala and Salzburg 2010–2016) (n=101, age 13.3 ± 2.8, m/f =50/51) were included (90 with overweight or obesity, 11 with normal weight). Standardized OGTT were performed and plasma glucose, glucagon and insulin concentrations assessed at baseline, 5, 10, 15, 30, 60, 90 and 120 minutes. Patients were grouped according to their glycemic state in six groups with normal glucose metabolism (NGM) and normal weight (NG-NW), NGM with obesity or overweight (NG-O), impaired glucose tolerance (IGT), impaired fasting glucose (IFG), IGT+IFG and T2D, and in two groups with NGM and impaired glucose metabolism (IGM), for statistical analysis.Results and conclusionGlucagon concentrations were elevated in young normoglycemic individuals with overweight or obesity (NG-O) compared to normoglycemic individuals with normal weight. Glucagon levels, fasting and dynamic, increased with progressing glycemic deterioration, except in IFG, where levels were comparable to those in NG-O. All glycemic groups showed an overall suppression of glucagon during OGTT. An initial increase of glucagon could be observed in T2D. In T2D, glucagon showed a strong direct linear correlation with plasma glucose levels during OGTT. Glucagon in adolescents, as in adults, may play a role in the disease progression of T2D.

Background gamma -aminobutyric acid (GABA), known as the main inhibitory neurotransmitter in the brain, exerts immunomodulatory functions by interaction with immune cells, including T cells. Metabolic programs of T cells are closely linked to their effector functions including proliferation, differentiation, and cytokine production. The physiological molecules glucose and insulin may provide environmental cues and guidance, but whether they coordinate to regulate GABA-mediated T cell immunomodulation is still being examined. Methods CD4 + T cells that were isolated from blood samples from healthy individuals and from patients with type 1 diabetes (T1D) were activated in vitro . We carried out metabolic assays, multiple proximity extension assay (PEA), ELISA, qPCR, immunoblotting, immuno fl uorescence staining, fl ow cytometry analysis, MS -based proteomics, as well as electrophysiology and live -cell Ca 2+ imaging. Findings We demonstrate that GABA-mediated reduction of metabolic activity and the release of in fl ammatory proteins, including IFN gamma and IL -10, were abolished in human CD4 + T cells from healthy individuals and patients with T1D when the glucose concentration was elevated above levels typically observed in healthy people. Insulin increased GABA A receptor -subunit rho 2 expression, enhanced the GABA A receptors -mediated currents and Ca 2+ in fl ux. GABA decreased, whereas insulin sustained, hexokinase activity and glycolysis in a glucose concentrationdependent manner. Interpretation These fi ndings support that metabolic factors, such as glucose and insulin, in fl uence the GABAmediated immunomodulation of human primary T cells effector functions.

Abstract Background Decreasing hyperinsulinemia is crucial in preventing laminitis in insulin dysregulated (ID) horses. Complementary pharmacological treatments that efficiently decrease postprandial hyperinsulinemia in ID horses are needed. Objectives Compare short‐term effects of canagliflozin vs placebo on glucose and insulin responses to an oral sugar test (OST) as well as the effects on body weight and triglyceride concentrations in horses with ID. Animals Sixteen privately‐owned ID horses. Methods A single‐center, randomized, double‐blind, placebo‐controlled, parallel design study. The horses were randomized (ratio 1:1) to either once daily PO treatment with 0.6 mg/kg canagliflozin or placebo. The study consisted of an initial 3‐day period for obtaining baseline data, a 3‐week double‐blind treatment period at home, and a 3‐day follow‐up period similar to the initial baseline period but with continued double‐blind treatment. Horses were subjected to an 8‐sample OST in the morning of the third day on both visits. Results Maximal geometric least square (LS) mean insulin concentration (95% confidence interval [CI]) during the OST decreased after 3 weeks of canagliflozin treatment compared with placebo (83.2; 55.4‐125.0 vs 215.2; 143.2‐323.2 μIU/mL). The geometric LS mean insulin response (insulin AUC0‐180) for canagliflozin‐treated horses was >66% lower compared with placebo. Least square mean body weight decreased by 11.1 (4‐18.1) kg and LS mean triglyceride concentrations increased by 0.99 (0.47‐1.5) mmol/L with canagliflozin treatment. Conclusions and Clinical Importance Canagliflozin is a promising drug for treatment of ID horses that requires future studies.

Introduction: Obesity is associated with chronic inflammation. Chronic inflammation has also been linked to insulin resistance and type 2 diabetes, metabolic associated fatty liver disease, and cardiovascular disease. Glucagon-like peptide-1 (GLP-1) receptor analogs (GLP-1RA) are clinically used to treat obesity, with known anti-inflammatory properties. How the GLP-1RA exenatide effects inflammation in adolescents with obesity is not fully investigated. Methods: Forty-four patients were randomized to receive weekly subcutaneous injections with either 2 mg exenatide or placebo for 6 months. Plasma samples were collected at baseline and at the end of the study, and 92 inflammatory proteins were measured. Results: Following treatment with exenatide, 15 out of the 92 proteins were decreased, and one was increased. However, after adjustment for multiple testing, only IL-18Rα was significantly lowered following treatment. Conclusions: Weekly injections with 2 mg of exenatide lowers circulating IL-18Rα in adolescents with obesity, which may be a potential link between exenatide and its anti-inflammatory effect in vivo. This contributes to exenatide’s pharmaceutical potential as a treatment for obesity beyond weight control and glucose tolerance, and should be further studied mechanistically.