Delayed diagnosis and treatment of extreme hypertriglyceridemia due to rejection of a lipemic sample

Most laboratories routinely determine haemolysis, icterus and lipemia indices to identify lipemic samples and reject potentially affected results. Hypertriglyceridemia is the most common cause of lipemia and severe hypertriglyceridemia (≥ 11.3 mmol/L) is a major risk factor of acute pancreatitis. A 56-year-old woman attended the outpatient clinic for a follow-up visit 1 month after a kidney transplantation. Her immunosuppressive therapy consisted of corticosteroids, cyclosporine, and mycophenolic acid. The routine clinical chemistry sample was rejected due to extreme lipemia. The comment “extreme lipemic sample” was added on the report, but the requesting physician could not be reached. The Cobas 8000 gave a technical error (absorption > 3.3) for the HIL-indices (L-index: 38.6 mmol/L) which persisted after high-speed centrifugation. The patient was given a new appointment 2 days later. The new sample was also grossly lipemic and gave the same technical error (L-index: 35.9 mmol/L). The second sample was manually diluted 20-fold after centrifugation to obtain a result for triglycerides within the measuring range (0.10–50.0 mmol/L). Triglycerides were 169.1 mmol/L, corresponding to very severe hypertriglyceridemia. This result was communicated to the nephrologist and the patient immediately recalled to the hospital. She received therapeutic plasma exchange the next day and did not develop acute pancreatitis. This case illustrates the delicate balance between avoiding the release of unreliable results due to lipemia and the risk of delayed diagnosis when results are rejected. Providing an estimate of the degree of hypertriglyceridemia might be preferable to rejecting the result

STE20 kinase TAOK3 regulates type 2 immunity and metabolism in obesity

Healthy adipose tissue (AT) contains ST2(+) Tregs, ILC2s, and alternatively activated macrophages that are lost in mice or humans on high caloric diet. Understanding how this form of type 2 immunity is regulated could improve treatment of obesity. The STE20 kinase Thousand And One amino acid Kinase-3 (TAOK3) has been linked to obesity in mice and humans, but its precise function is unknown. We found that ST2(+) Tregs are upregulated in visceral epididymal white AT (eWAT) of Taok3(-/-) mice, dependent on IL-33 and the kinase activity of TAOK3. Upon high fat diet feeding, metabolic dysfunction was attenuated in Taok3(-/-) mice. ST2(+) Tregs disappeared from eWAT in obese wild-type mice, but this was not the case in Taok3(-/-) mice. Mechanistically, AT Taok3(-/-) Tregs were intrinsically more responsive to IL-33, through higher expression of ST2, and expressed more PPAR & gamma; and type 2 cytokines. Thus, TAOK3 inhibits adipose tissue Tregs and regulates immunometabolism under excessive caloric intake. Maes et al. reveal an unexpected role of TAOK3 in regulating ST2(+) regulatory T cells in mouse adipose tissue. Absence of TAOK3 sustains Tregs in obesity and improves metabolic dysfunction

AURORA : bariatric surgery registration in women of reproductive age : a multicenter prospective cohort study

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AURORA : bariatric surgery registration in women of reproductive age : a multicenter prospective cohort study

Background: The expansion of the obesity epidemic is accompanied with an increase in bariatric procedures, in particular in women of reproductive age. The weight loss induced by the surgery is believed to reverse the negative impact of overweight and obesity on female reproduction, however, research is limited to in particular retrospective cohort studies and a growing number of small case-series and case-(control) studies. Methods/design: AURORA is a multicenter prospective cohort study. The main objective is to collect long-term data on reproductive outcomes before and after bariatric surgery and in a subsequent pregnancy. Women aged 18-45 years are invited to participate at 4 possible inclusion moments: 1) before surgery, 2) after surgery, 3) before 15 weeks of pregnancy and 4) in the immediate postpartum period (day 3-4). Depending on the time of inclusion, data are collected before surgery (T1), 3 weeks and 3, 6, 12 or x months after surgery (T2-T5) and during the first, second and third trimester of pregnancy (T6-T8), at delivery (T9) and 6 weeks and 6 months after delivery (T10-T11). Online questionnaires are send on the different measuring moments. Data are collected on contraception, menstrual cycle, sexuality, intention of becoming pregnant, diet, physical activity, lifestyle, psycho-social characteristics and dietary supplement intake. Fasting blood samples determine levels of vitamin A, D, E, K, B-1, B-12 and folate, albumin, total protein, coagulation parameters, magnesium, calcium, zinc and glucose. Participants are weighted every measuring moment. Fetal ultrasounds and pregnancy course and complications are reported every trimester of pregnancy. Breastfeeding is recorded and breast milk composition in the postpartum period is studied. Discussion: AURORA is a multicenter prospective cohort study extensively monitoring women before undergoing bariatric surgery until a subsequent pregnancy and postpartum period

The INNODIA Type 1 Diabetes Natural History Study: a European cohort of newly diagnosed children, adolescents and adults

Aims/hypothesis: Type 1 diabetes is an heterogenous condition. Characterising factors explaining differences in an individual’s clinical course and treatment response will have important clinical and research implications. Our aim was to explore type 1 diabetes heterogeneity, as assessed by clinical characteristics, autoantibodies, beta cell function and glycaemic outcomes, during the first 12 months from diagnosis, and how it relates to age at diagnosis. Methods: Data were collected from the large INNODIA cohort of individuals (aged 1.0–45.0 years) newly diagnosed with type 1 diabetes, followed 3 monthly, to assess clinical characteristics, C-peptide, HbA1c and diabetes-associated antibodies, and their changes, during the first 12 months from diagnosis, across three age groups: <10 years; 10–17 years; and ≥18 years. Results: The study population included 649 individuals (57.3% male; age 12.1±8.3 years), 96.9% of whom were positive for one or more diabetes-related antibodies. Baseline (IQR) fasting C-peptide was 242.0 (139.0–382.0) pmol/l (AUC 749.3 [466.2–1106.1] pmol/l × min), with levels increasing with age (p<0.001). Over time, C-peptide remained lower in participants aged <10 years but it declined in all age groups. In parallel, glucose levels progressively increased. Lower baseline fasting C-peptide, BMI SD score and presence of diabetic ketoacidosis at diagnosis were associated with lower stimulated C-peptide over time. HbA1c decreased during the first 3 months (p<0.001), whereas insulin requirement increased from 3 months post diagnosis (p<0.001). Conclusions/interpretation: In this large cohort with newly diagnosed type 1 diabetes, we identified age-related differences in clinical and biochemical variables. Of note, C-peptide was lower in younger children but there were no main age differences in its rate of decline. Graphical Abstract

Association of Adipose tissue inflammation with histologic severity of nonalcoholic fatty liver disease

BACKGROUND & AIMS : The prevalence of nonalcoholic fatty liver disease (NAFLD) has increased with the obesity pandemic. We analyzed the transcriptional profiles of subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT), and phenotypes and functional characteristics of adipocyte tissue macrophages (ATMs), in obese patients undergoing bariatric surgery. METHODS : We collected anthropometric data; plasma samples; and SAT, VAT, and liver tissues from 113 obese patients undergoing bariatric surgery at academic hospitals in Europe (Antwerp and Leuven) and South Africa. Based on clinical and histologic features, patients were assigned to the following groups: obese, NAFLD, nonalcoholic steatohepatitis (NASH), or NASH with fibrosis. Microarray analyses were performed to identify genes expressed differentially among groups. We measured levels of cytokines and chemokines in plasma samples and levels of RNAs in adipose tissues by quantitative reverse-transcription polymerase chain reaction. ATMs were isolated from patients and 13 lean individuals undergoing cholecystectomy (controls), analyzed by flow cytometry, and cultured; immunophenotypes and levels of cytokines and chemokines in supernatants were determined. RESULTS : We observed increased expression of genes that regulate inflammation in adipose tissues from patients with NAFLD and NASH; expression of these genes increased as disease progressed from NAFLD to NASH. We found 111 genes associated with inflammation that were expressed differentially between VAT and SAT. Serum levels of interleukin 8, chemokine (C-C motif) ligand 3, and tumor necrosis factor-a correlated with liver inflammation and NAFLD activity score. We developed 2 models that could be used to determine patients’ liver histology based on gene expression in VAT and SAT. Flow cytometry showed increased proportions of CD11cþCD206þ and CCR2þ macrophages in VAT from patients with NASH, and supernatants of cultured macrophages had increased levels of cytokines and chemokines compared with controls. CONCLUSIONS : VAT and SAT from patients with NAFLD and NASH have an increased expression of genes that regulate inflammation, and ATM produce increased levels of inflammatory cytokines, compared with adipose tissues from controls. We identified an expression profile of 5 genes in SAT that accurately predict liver histology in these patients. Transcript profiling: accession numbers: GSE58979 and GSE59045.Schalk van der Merwe, Chantal Mathieu, Frederik Nevens, David Cassiman, and Sven Francque are recipients of the Flanders fund for scientific research (FWO klinisch mandaat), and Hannelie Korf is a recipient of the FWO postdoctoral mandate. Research at the Department of Endocrinology, Diabetology and Metabolism and the Department of Gastroenterology and Hepatology of the Antwerp University Hospital (Belgium) was supported by the European Union: FP6 (HEPADIP contract LSHM-CT-2005-018734) and FP7-HEALTH (RESOLVE no. 305707). Supported by a fellowship from the South African Gastroenterology Association and a scholarship from the European Association for the Study of the Liver (J.d.P.). This research also was supported by a research grant from the Gastro foundation of South Africa. The authors specifically acknowledge the support of Dr. Chris Kassianides. Also funded in part by a grant from the Deutsche Forschungsgemeinschaft DFG-SFB 1052/1: Obesity Mechanisms (projects A04) and by the Helmholtz Alliance Imaging and Curing Environmental Metabolic Disease through the Initiative and Networking Fund of the Helmholtz Association (M.G.).http://www.journals.elsevier.com/gastroenterology2016-09-30hb2016Internal Medicin

Genotype-stratified treatment for monogenic insulin resistance: a systematic review

This is the final version. Available from Nature Research via the DOI in this record. Data availability: All data used in this review is available from publicly available and herein referenced sources. A list of included studies is provided in Supplementary Data 1. All data generated or analyzed during this study are included in this published article and its supplementary information files. Source data for the figures are available as Supplementary Data 2.BACKGROUND: Monogenic insulin resistance (IR) includes lipodystrophy and disorders of insulin signalling. We sought to assess the effects of interventions in monogenic IR, stratified by genetic aetiology. METHODS: Systematic review using PubMed, MEDLINE and Embase (1 January 1987 to 23 June 2021). Studies reporting individual-level effects of pharmacologic and/or surgical interventions in monogenic IR were eligible. Individual data were extracted and duplicates were removed. Outcomes were analysed for each gene and intervention, and in aggregate for partial, generalised and all lipodystrophy. RESULTS: 10 non-randomised experimental studies, 8 case series, and 23 case reports meet inclusion criteria, all rated as having moderate or serious risk of bias. Metreleptin use is associated with the lowering of triglycerides and haemoglobin A1c (HbA1c) in all lipodystrophy (n = 111), partial (n = 71) and generalised lipodystrophy (n = 41), and in LMNA, PPARG, AGPAT2 or BSCL2 subgroups (n = 72,13,21 and 21 respectively). Body Mass Index (BMI) is lowered in partial and generalised lipodystrophy, and in LMNA or BSCL2, but not PPARG or AGPAT2 subgroups. Thiazolidinediones are associated with improved HbA1c and triglycerides in all lipodystrophy (n = 13), improved HbA1c in PPARG (n = 5), and improved triglycerides in LMNA (n = 7). In INSR-related IR, rhIGF-1, alone or with IGFBP3, is associated with improved HbA1c (n = 17). The small size or absence of other genotype-treatment combinations preclude firm conclusions. CONCLUSIONS: The evidence guiding genotype-specific treatment of monogenic IR is of low to very low quality. Metreleptin and Thiazolidinediones appear to improve metabolic markers in lipodystrophy, and rhIGF-1 appears to lower HbA1c in INSR-related IR. For other interventions, there is insufficient evidence to assess efficacy and risks in aggregated lipodystrophy or genetic subgroups.Wellcome TrustWellcome Trus

New Targets in Migraine Therapy

Migraine is a common neurovascular disorder which ranks among the world s most disabling medical conditions and has been estimated to be the mos t costly neurological disease in the European Community. Although knowle dge of the main molecular players is still incomplete, intensive researc h during the last decade has led to conceptual changes in our understand ing of migraine headache (Chapter 1). Current acute and prophylactic treatment options are far from optimal and consequently improved treatme nts based on the new insights in the pathophysiology of migraine are des perately needed. In the present thesis, a number of clinical pharmacological experimental methods are used to evaluate emerging new targets for the treatment of migraine in vivo in humans (Chapter 2). The main aim is to link fi ndings from basic science to their clinical applications quickly, exempl ifying the relevance of clinical pharmacology and its contribution to tr anslational medicine and the explorative phase of clinical drug developm ent. Our research focuses on three promising new treatment strategies: ( 1) calcitonin gene-related peptide (CGRP) receptor antagonism (Part I), (2) nitric oxide synthase (NOS) inhibition (Part II) and (3) inter vention in the endocannabinoid system (Part III). In Part I of the thesis, we translate a non-invasive pharmacodynamic model used in animals to allow rapid evaluation of CGRP receptor antagon ists into an in vivo human model. As the selectivity of CGRP receptor antagonists is known to differ between species this translational step is crucial for investigating proof-of-mechanism of CGRP antagonists in h umans. The model uses topical application of capsaicin to the skin of he althy volunteers to provoke CGRP release leading to an increase in derma l blood flow (DBF) which can be measured by Laser Doppler perfusion imag ing. In a first exploratory study in 12 healthy men, we identify the dos e of capsaicin needed to produce a robust and reproducible dermal blood flow (DBF) response measured by Laser Doppler perfusion imaging after to pical application to the human forearm (Chapter 3). Simultaneously, t he influence of the forearm location on the capsaicin response is assess ed. In the proximal forearm of healthy volunteers a 1000 µg capsaicin do se elicits a reproducible response in both proximal sites at the 30-minu tes time point. Subsequently, a reproducibility study is performed in 21 healthy men to assess the within-subject arm-to-arm and period-to-perio d reproducibility of the forearm DBF response to capsaicin using the 100 0 µg capsaicin dose and the proximal forearm locations. In this second s tudy, the data of 11 subjects with an increase in forearm blood flow fro m baseline &#8805; 100 % (i.e. responders) during both study visits are included in a reproducibility analysis. The arm-to-arm reproducibility o f the DBF after capsaicin application is almost perfect and based on the within-subject standard deviation the sample-size needed to detect diff erences in capsaicin-induced DBF increase between arms are calculated. Subsequently, we characterize and indentify the mediators involved in th e dermal vasodilation induced by capsaicin application to the human skin (Chapter 4). To assess the involvement of CGRP, the CGRP receptor an tagonist CGRP8-37 is infused into the brachial artery of one arm. In thi s way, the non-infused arm is not treated and serves as the control arm. The involvement of nitric oxide (NO) and prostaglandins in capsaicin-in duced DBF increase are evaluated in a similar way by infusing the non-se lective NO synthase inhibitor, NG-monomethyl-L-arginine monoacetate (L-N MMA), and the cyclo-oxygenase inhibitor indomethacin, respectively. To a ssess to involvement of substance P (SP) the potent SP antagonist aprepi tant is given orally, and capsaicin-induced vasodilation is measured pre dose on the left arm and postdose (Tmax) on the right arm. The main find ing from this study is that capsaicin-induced vasodilation in the human skin is to a large extent antagonized by CGRP8-37, but not by L-NMMA, ap repitant or indomethacin. Therefore CGRP seems to be the major mediator involved in this vasodilatory response to capsaicin. In addition, CGRP8- 37 does not affect resting DBF, indicating that CGRP does not play a rol e in maintaining basal DBF. The proposed model allows the assessment of the pharmacodynamic efficacy of CGRP receptor antagonists in a small gro up of healthy subjects and may guide the finding of a clinical effective dose. In Part I of the thesis, we also use tools from clinical pharmacology to assess the vascular effects and safety of CGRP receptor antagonists. Data in animals suggest that administration of nitroglycerin (NTG), whi ch is widely used in the treatment of cardiac ischemic pain (i.e. angina pectoris), causes release of CGRP in the cardiovascular system resultin g in coronary artery vasodilatation. We therefore want to investigate th e effect of CGRP receptor antagonism on response to NTG in vivo in hu mans (Chapter 5). We conduct a study in 22 healthy men and assessed v ascular changes using non-invasive techniques, including ultrasound meas urement of the brachial artery diameter and systolic pulse contour analy sis. By administering NTG following a high clinical dose of the potent C GRP receptor antagonist, telcagepant, we demonstrate that CGRP antagonis m has no clinically relevant effect on NTG-induced hemodynamic changes i n healthy men. Nor is there a measurable vasoconstrictor effect of telca gepant as such in both the central and peripheral vascular beds. These r esults support the favourable cardiovascular safety profile of CGRP rece ptor antagonists, especially when compared to triptans. In Part II of the thesis, the involvement of NO synthase activity in the pathogenesis of migraine is investigated by measuring associated bio markers of systemic NO production. In clinical pharmacology, the measure ment of biomarkers is used to identify potential treatment targets. Late r, the same biomarkers can often be used as surrogates to evaluate pre liminary clinical efficacy of newly developed compounds. There is abunda nt evidence for the involvement of NO in the pathogenesis of migraine. N O induces delayed headaches in migraineurs which resemble the characteri stics of spontaneous migraine attacks and the non-selective NO synthase inhibitor L-NMMA provides pain relief to patients experiencing a spontan eous migraine headache. In a first study, we assess whether migraine patients display a chronic NOS hyperactivity (Chapter 6). Based on the results of a reproducibil ity study in 12 healthy volunteers, we set up a study to compare NO prod uction before and following a loading dose of L-arginine between 20 migr aine patients, outside of a migraine attack, and 20 healthy volunteers b y measuring biomarkers associated with the L-arginine/NO conversion (i.e . nasal and exhaled NO, plasma L-citrulline and L-ariginine and urinary excretion of nitrates and cyclic guanosine monophosphate (cGMP)). The re sults of this study do not support the idea of a generalized increase in NOS activity in migraine patients outside of a migraine attack. On the contrary, following L-arginine infusion, the increase in plasma L-citrul line levels and urinary excretion of nitrite/nitrate and cGMP is smaller in migraine patients compared to healthy volunteers. The latter might i ndicate dysfunction of endothelial NOS in migraine patients and adds to the ongoing discussion about migraine being an endotheliopathy leading t o decreased endothelial NOS. The second study in Part II of the thesis investigates the pharmacoki netic and pharmacodynamic properties of GW273629, a potent inducible NOS (iNOS) inhibitor, in 15 migraine patients both during and outside of an acute migraine attack (Chapter 7). As in the first study, systemic N O production is evaluated by measuring biomarkers associated with the L- arginine/NO pathway (i.e. nasal and exhaled NO, plasma nitrate and plasm a 3-nitrotyrosine). This allows to assess the effect of migraine headach e and selective inducible NO synthase inhibition upon systemic NO produc tion. Although the early absorption of orally administered GW273629 is d elayed during a migraine attack, the time-course of iNOS inhibition, as assessed by nasal and exhaled NO, is similar during and outside of a mig raine attack. So while migraine headache changes the pharmacokinetic pro perties of GW273629, its pharmacodynamic profile remains unaffected . Re sults from this study suggest that migraine headache might be associated with excess systemic NO production, but strong iNOS inhibition by GW273 629 has no beneficial effect in this small open label study. Recent larg e clinical trials have also failed to show any efficacy of potent iNOS i nhibitors, both in the acute and the prophylactic treatment of a migrain e headache. In Part III of the thesis, we use recent advances in neuroimaging to evaluate putative changes in the endocannabinoid system associated with migraine. In vivo investigation of the type 1 cannabinoid receptor (CB1R ) has become possible with the development of positron emission tomograp hy (PET) radioligands. We compare cerebral CB1R availability between 20 female migraineurs outside the acute phase of a migraine attack and 18 h ealthy women. Migraine patients present an increase in CB1R availability in all brain regions, especially in the cingulo-frontal cortex and the limbic system which presumably exert top-down influences to modulate pai n. This is in accordance with recent findings suggesting that a failure of the endogenous endocannabinoid system might play a role in the pathop hysiology of migraine and its tendency to progress to chronic migraine. This study is another illustration of how clinical pharmacology uses a l arge variety of tools to identify potential treatment targets. Our findi ng that there exists a dysregulation in the endocannabinoid system in mi graine patients should encourage further investigation into cannabinoid signalling and might ultimately lead to pharmacological advances in the treatment of migraine. General conclusion This thesis presents a number of tools from clinical pharmacology which allow to both identify and facilitate clinical development of new pharma cological migraine treatments. Whereas preclinical research remains esse ntial in the quest for more efficient and safer migraine therapy, our re sults clearly underline the need for human models as there seems to be l arge variability with regard to the mediators involved in the pathophysi ological processes associated with the development of a migraine headach e. In addition, the mere nature of migraine renders the development of a nimal research models difficult and therefore their validity remains unc ertain. Whereas CGRP receptor antagonists for migraine treatment will soon appea r on the market, the future of NOS inhibition and modulation of the endo cannabinoid system remains uncertain. Non-selective NOS inhibition is ef fective in the treatment of migraine but unattractive because of cardiov ascular safety concerns. Our results favour the involvement of nNOS in t he pathogenesis of migraine above iNOS and eNOS but clinical data are la cking at this moment. Finally, therapeutic intervention in the endocanna binoid system does seem attractive, but the recent withdrawal from the m arket of cannabinoid receptor antagonists clearly illustrates that cauti on is warranted with regard to central nervous system side effects.status: publishe

Soft and hard rapid tools for polymer and metal casting/moulding

status: publishe