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Objective: Vaso-occlusive crisis (VOC) is a common clinical manifestation of sickle cell anemia (SCA) and is associated with increased proinflammatory mediators. Copeptin is the C-terminal part of the prohormone for provasopressin and seems clinically relevant in various clinical conditions. Right ventricular (RV) dysfunction significantly appears in SCA patients due to pulmonary hypertension. This study aimed to investigate the association of copeptin levels in VOC patients and evaluate RV dysfunction. Materials and Methods: A total of 108 patients were enrolled in the study. Twenty-eight SCA patients in steady state (30.2 ± 0.9 years), 25 SCA patients in VOC (36.8 ± 11.8 years), and 55 healthy individuals (31.9 ± 9.4 years) with HbAA genotype were included. Clinical, echocardiographic, and laboratory data were recorded. ELISA was used for the determination of serum levels of copeptin. Results: VOC patients had significantly higher copeptin level compared both with controls and SCA subjects in steady state (22.6 ± 13.0 vs. 11.3 ± 5.7 pmol/L, 22.6 ± 13.0 vs. 12.4 ± 5.8 pmol/L, p = 0.009 for both). Additionally, the copeptin level was significantly higher in SCA patients with RV dysfunction than those without RV dysfunction (23.2 ± 12.2 vs. 15.3 ± 9.5 pmol/L, p = 0.024). Multiple logistic regression analysis revealed that high-sensitivity C-reactive protein and copeptin levels were found to be associated with VOC. Conclusion: This study showed that copeptin and hs-CRP levels were increased in patients with VOC, and it was found that RV dysfunction was more common in SCA patients with VOC than in the control group. Copeptin can be considered for use as a potential biomarker in predicting VOC crisis in SCA patients and in the early detection of patients with SCA who have the potential to develop RV dysfunction.

Aim. To compare the effectiveness of the angiotensin-converting enzyme inhibitor enalapril and the angiotensin II receptor antagonist valsartan in patients with heart failure with mid-range ejection fraction (HFmrEF) from the standpoint of the effect on the clinical picture, echocardiographic parameters and the level of the N-terminal fragment of the prohormone brain-type sodium (NT-proBNP). Methods. 110 patients with heart failure with mid-range ejection fraction were included in the study based on the City Clinical Hospital named after N.I. Pirogov of Orenburg between 2018 and 2020. All patients were divided into two randomized groups. Patients of the first group (n=55) were prescribed enalapril, the second group (n=55) valsartan. Each patient was followed up for 1 year. The six-minute walk test, NT-pro-brain natriuretic peptide level, echocardiography parameters were assessed in dynamics. Statistical analysis was performed by using Statistica 10.0 software, ShapiroWilk, MannWhitney, Wilcoxon tests. Results. During the year of observation in both groups, there was a significant decrease in the functional class of chronic heart failure (p 0.005) without a statistical difference between the groups (p=0.251). The distance during the six-minute walk test increased from 350 (310400) m to 490 (420530) m (p 0.001) in the first group, from 360 (330400) m to 510 (450520) m (p 0.001) in the second group, also without significant differences (p=0.361). The NT-pro-brain natriuretic peptide level decreased from 491 (410610) pg/ml to 286 (187350) pg/ml (p 0.001) in the first group, and from 446 (376534) pg/ml to 210 (143343) pg/ml (p 0.001) in the second, with a more significant change in the second group (p=0.020). The dynamics of echocardiography parameters were comparable in the groups (p 0.05), while ejection fraction normalized in 89.1% of patients received enalapril and 92.7% of patients received valsartan. Conclusion. The efficacy of enalapril and valsartan in heart failure with mid-range ejection fraction is comparable in its effect on the clinical picture and echocardiography parameters with a more pronounced decrease in NT-pro-brain natriuretic peptide when taking valsartan during a year of follow up.

Improvements in both liquid chromatography (LC) and mass spectrometry (MS) instrumentation have greatly enhanced proteomic and small molecule metabolomic analysis in recent years. Less focus has been on the improved capability to detect and quantify small bioactive peptides, even though the exact sequences of the peptide species produced can have important biological consequences. Endogenous bioactive peptide hormones, for example, are generated by the targeted and regulated cleavage of peptides from their prohormone sequence. This process may include organ specific variants, as proglucagon is converted to glucagon in the pancreas but glucagon-like peptide-1 (GLP-1) in the small intestine, with glucagon raising, whereas GLP-1, as an incretin, lowering blood glucose. Therefore, peptidomics workflows must preserve the structure of the processed peptide products to prevent the misidentification of ambiguous peptide species. The poor in vivo and in vitro stability of peptides in biological matrices is a major factor that needs to be considered when developing methods to study them. The bioinformatic analysis of peptidomics data sets requires the inclusion of specific post-translational modifications, which are critical for the function of many bioactive peptides. This review aims to discuss and contrast the various extraction, analytical, and bioinformatics approaches used for human peptidomics studies in a multitude of matrices.

Aims Risk stratification in patients with advanced chronic heart failure (HF) is an unmet need. Circulating microRNA (miRNA) levels have been proposed as diagnostic and prognostic biomarkers in several diseases including HF. The aims of the present study were to characterize HF‐specific miRNA expression profiles and to identify miRNAs with prognostic value in HF patients. Methods and results We performed a global miRNome analysis using next‐generation sequencing in the plasma of 30 advanced chronic HF patients and of matched healthy controls. A small subset of miRNAs was validated by real‐time PCR (P

Background: Thyroxine (T4) is generally considered to be a prohormone that requires conversion to triiodothyronine (T3) to exert biological activity. Although evidence suggests that T4 has intrinsic activity, it is questionable if this activity has any physiological relevance. Methods: To answer this question, triple knockout (KO) mice (Triples) that cannot express the types 1 (D1) and 2 (D2) deiodinase and the Pax8 genes were generated. Thus, they lack a thyroid and cannot convert T4 to T3. Triples were injected on alternate days with either vehicle or physiological doses of T4, T3, or T3+T4 from postnatal days 2–14. They were euthanized at P15, and RNA-seq was employed to profile gene expression in the liver. In another experiment, Pax8KO mice were injected with T3, T4, or T4+T3, and growth rate and survival to P84 were determined. Results: The growth retardation of Triples was not improved by either T3 or T4 alone but was significantly improved by T4+T3. In the liver, T4 significantly regulated the expression of genes that were also regulated by T3, but the proportion of genes that were negatively regulated was higher in mice treated with T4 than in mice treated with T3. Treatment with T4+T3 identified genes that were regulated synergistically by T3 and T4, and genes that were regulated only by T4+T3. Analysis of these genes revealed enrichment in mechanisms related to cell proliferation and cholesterol physiology, suggesting a unique contribution of T4 to these biological functions. Pax8KO mice all survived to P84 when injected with T4 or T4+T3. However, survival rate with T3 was only 50% and 10% at 3.5 and 12 weeks of life, respectively. Conclusions: T4 has intrinsic activity in vivo and is critical for survival and growth. At a physiological level, T4 per se can upregulate or downregulate many T3 target genes in the neonatal liver. While most of these genes are also regulated by T3, subsets respond exclusively to T4 or demonstrate enhanced or normalized expression only in the presence of both hormones. These studies demonstrate for the first time a complex dependency on both T4 and T3 for normal mammalian growth and development.

Vitamin D can be obtained from diet, direct sunlight, or supplementation. The most common form is synthesized in the skin after exposure to ultraviolet B radiation. Nevertheless, the thought is that vitamin D is more of a multifunctional hormone or prohormone. This is because vitamin D plays contributes to many processes in the body. Calcitriol has been shown to have enhancing effects on the immune system, the cardiovascular system, the endocrine system, and other metabolic pathways. There is evidence that vitamin D has also a role in depression, pain, and cancer.

Proopiomelanocortin (POMC) neurons contribute to the regulation of many physiological processes; the majority of which have been attributed to the release of peptides produced from the POMC prohormone such as α-MSH, which plays key roles in food intake and metabolism. However, it is now clear that POMC neurons also release amino acid transmitters that likely contribute to the overall function of POMC cells. Recent work indicates that constitutive deletion of these transmitters can affect metabolic phenotypes, but also that the expression of GABAergic or glutamatergic markers changes throughout development. The goal of the present study was to determine whether the release of glutamate or GABA from POMC neurons in the adult mouse contributes notably to energy balance regulation. Disturbed release of glutamate or GABA specifically from POMC neurons in adult mice was achieved using a tamoxifen-inducible Cre construct ( Pomc-CreERT2) expressed in mice also carrying floxed versions of Slc17a6 (vGlut2) or Gad1 and Gad2, encoding the vesicular glutamate transporter type 2 and GAD67 and GAD65 proteins, respectively. All mice in the experiments received tamoxifen injections, but control mice lacked the tamoxifen-inducible Cre sequence. Body weight was unchanged in Gad1- and Gad2- or vGlut2-deleted female and male mice. Additionally, no significant differences in glucose tolerance or refeeding after an overnight fast were observed. These data collectively suggest that the release of GABA or glutamate from POMC neurons in adult mice does not significantly contribute to the metabolic parameters tested here. In light of prior work, the data also suggest that amino acid transmitter release from POMC cells may contribute to separate functions in the adult versus the developing mouse.

Bronchopulmonary dysplasia (BPD) is a combined pulmonary vascular and parenchymal disease, representing the most common cause of chronic lung disease (CLD) in infancy. Pulmonary hypertension (PH) is frequently associated with BPD and—if persistent—substantially increases mortality. We report on a 4-month-old, former preterm infant with BPD, severe PH and right heart failure who greatly and rapidly improved clinical status and right ventricular (RV) function by means of blood biomarkers [N-terminal prohormone of brain natriuretic peptide (NT-pro-BNP), cardiac troponin T] and transthoracic echocardiography, following the addition of spironolactone and hydrochlorothiazide to the treatment regimen.

Atrial Fibrillation (AF) is a growing public health issue, associated with significant morbidity and mortality. In addition to pharmacological therapy, catheter ablation is an effective strategy in restoring and maintaining sinus rhythm. However, ablation is not without risk, and AF recurs in a significant proportion of patients. Non-invasive, easily accessible markers or indices that could stratify patients depending on the likelihood of a successful outcome following ablation would allow us to select the most appropriate patients for the procedure, reducing the AF recurrence rate and exposure to potentially life-threatening risks. : There has been much attention paid to Brain Natriuretic Peptide (BNP) and N-Terminal prohormone of Brain Natriuretic Peptide (NT-proBNP) as possible predictive markers of successful ablation. Several studies have demonstrated an association between higher pre-ablation levels of these peptides, and a greater likelihood of AF recurrence. Therefore, there may be a role for measuring brain natriuretic peptides levels when selecting patients for catheter ablation.

Aims Efforts to improve access to antiretroviral therapy (ART) have shifted morbidity and mortality among persons living with HIV (PLWH) from AIDS to non‐communicable diseases, such as cardiovascular disease (CVD). However, contemporary data on CVD among PLWH in sub‐Saharan Africa in the current ART era are lacking. The aim of this study was to assess the burden of cardiac stress among PLWH in South Africa via measurement of N‐terminal prohormone of brain natriuretic peptide (NT‐proBNP). Methods and results NT‐proBNP was measured at baseline in 224 PLWH enrolled in a sub‐study of a tuberculosis vaccine trial in Khayelitsha township near Cape Town, South Africa. Thresholds were applied at the assay's limit of detection (≥137 pg/mL) and a level indicative of symptomatic heart failure in the acute setting (>300 pg/mL). Mean (SD) age of participants was 39 (6) years, 86% were female, and 19% were hypertensive. Mean (SD) duration of HIV diagnosis was 8.3 (3.9) years and CD4 + count was 673 (267) with 79% prescribed ART for a duration of 5.6 (2.7) years. Thirty‐one percent of participants had NT‐proBNP > 300 pg/mL. Elevated vs. undetectable NT‐proBNP level was associated with older age (P = 0.04), no ART (P = 0.03), and higher plasma tumour necrosis factor‐α (P = 0.01). Conclusions Among South African PLWH largely free of known CVD and on ART with high CD4 + counts and few comorbidities, we observed a high proportion with elevated NT‐proBNP levels, suggesting the burden of cardiac stress in this population may be high. This observation underscores the need for more in‐depth research, including the current effect of HIV on heart failure risk among a growing ART‐treated population in sub‐Saharan Africa.

BACKGROUND Heart failure is characterized by a tissue damage that progressively leads to mechanical cardiac dysfunction and remodeling. A recent investigation showed that α-1 antitripsin, an antiprotease, able to inhibit metalloproteinases, provides prognostic information about heart failure and mortality postacute myocardial infarction. Therefore, we conducted a study to establish if α-1 antitrypsin (AAT) could be considered a marker of severity of heart failure. METHODS A total of 182 heart failure patients (Group 1) were enrolled and AAT values were compared with controls (Group 2). RESULTS In Group 1 a significant increment of AAT levels respect to Group 2 was observed (P

Cigarettes are the biggest health problem around the world. Nicotine in cigarettes has been proved insulin receptor resistance and can reduce insulin secretion in pancreatic β cells, which causes an increase in blood glucose levels. One of the efforts to reduce blood glucose levels is to consume vitamin D3. Vitamin D3 is a prohormone that plays a role in metabolic functions in cells and repair functions of cells. Purpose: to know the effect of vitamin D3 toward glucose blood level of male Rattus norvegicus Wistar exposed by cigarette smoke. Methods: This research conducted with pure experimental pre- and post-test with control group design. The samples used were 24 male rats. The sample divided into four groups, including K1, K2, K3, and P1. K1, which is not exposed by cigarette smoke and not given of vitamin D3. K2 is a group exposed by cigarette smoke but not given vitamin D3. K3 is a group that is not exposed by cigarette smoke but given vitamin D3 a dose of 0.2 µgr /head. P1 is a group exposed by cigarette smoke and given a vitamin D3 dose of 0.2 µgr /head. Results: Paired T-test showed increased blood glucose levels that are meaningful (p

Background : Multiple Sclerosis is a neurologic disease that effect Central Nervous System(CNS). Vitamin D is a liposoluble prohormone. It is thought that vitamin D deficiency is responsible for Lots of diseases which includes MS . In this study, we aimed to show the relationship between serum vitamin D level and MS. Objective : In our research we wanted to show the serum vitamin D levels in MS patients. Methods : We included 106 patients (76 female, 30 male) to our study. Participants were divided into two groups as patient and control group. Serum vitamin D levels were evaluated. We excluded patients with known liver or renal disease. Liver enzymes and blood creatinine of the patients included in the study were normal. Results: There was a statistically significant difference when comparing two groups of serum vitamin D results. Serum vitamin D levels were lower in the control group. No correlation was found between vitamin D and age. There was no correlation between vitamin D and gender. According to the vitamin D level reference interval in the MS group, 28 patients had deficiency, 17 patients had failure, and 12 patients were normal. In the control group, according to the vitamin D level reference range, 35 patients had deficiency, 11 patients had failure and 3 patients were normal. Conclusion: In this study conducted with limited facilities, lower vitamin D levels in the control group suggested that vitamin D levels may be low when the population is examined in general. Vitamin D support throughout the population may reduce the prevalence of other diseases thought to be caused by vitamin D deficiency as well as MS.

Neuropeptides are a diverse class of signalling molecules in metazoans. They occur in all animals with a nervous system and also in neuron-less placozoans. However, their origin has remained unclear because no neuropeptide shows deep homology across lineages and none have been found in sponges. Here, we identify two neuropeptide precursors, phoenixin and nesfatin, with broad evolutionary conservation. By database searches, sequence alignments and gene-structure comparisons we show that both precursors are present in bilaterians, cnidarians, ctenophores and sponges. We also found phoenixin and a secreted nesfatin precursor homolog in the choanoflagellate Salpingoeca rosetta. Phoenixin in particular, is highly conserved, including its cleavage sites, suggesting that prohormone processing occurs also in choanoflagellates. In addition, based on phyletic patterns and negative pharmacological assays we question the originally proposed GPR-173 (SREB3) as a phoenixin receptor. Our findings indicate that signalling by secreted neuropeptide homologs has pre-metazoan origins and thus evolved before neurons.

Aging is associated with the development of metabolic disorders, including insulin resistance and hypertension. Young mice that are negative for the neuroendocrine prohormone Chromogranin A (CgA knockout, CgA-KO) display two opposite aging phenotypes: hypertension but heightened insulin sensitivity. We determined these phenotypes in aging CgA mice. In comparison, aging wild-type (WT) mice gradually lost glucose tolerance and insulin sensitivity. Moreover, while aging WT mice had increased inflammation with higher plasma TNF-α, IFN-γ and CCL2 and increased mitochondrial fission, these phenotypes were the opposite in aging CgA-KO mice. CgA-KO mice also showed increased expression of mitochondrial and nuclear-encoded complex I genes, implying that they were healthier than WT mice. Most intriguingly, the hypertension in CgA-KO mice was spontaneously reversed with aging. Supplementation of CgA-KO mice with pancreastatin, a hyperglycemic peptide produced from CgA by proteolysis, increased both blood glucose levels and blood pressure, implicating hyperglycemia, and hypertension. We conclude that age-related insulin resistance and hypertension are caused by CgA.

Autologous pancreatic islet transplantation is an established therapy for patients with chronic pancreatitis. However, the long-term transplant outcomes are modest. Identifying indicators of graft function will aid the preservation of transplanted islets and glycemic control. To this end, we analyzed beta cell prohormone peptide levels in a retrospective cohort of total pancreatectomy autologous islet transplant patients (n=28). Proinsulin-to-C-peptide (PI/C) and proIAPP-to-total IAPP (proIAPP/IAPP) ratios measured at 3 months post-transplant were significantly higher in patients who remained insulin dependent at 1 year follow-up. In a mouse model of human islet transplantation, recipient mice that later became hyperglycemic displayed significantly higher PI/C ratios than mice that remained normoglycemic. Histological analysis of islet grafts showed reduced insulin- and proinsulin-positive area, but elevated glucagon-positive area in grafts that experienced greater secretory demand. Increased prohormone convertase 1/3 was detected in glucagon-positive cells, and glucagon-like peptide 1 (GLP-1) area was elevated in grafts from mice that displayed hyperglycemia or elevated plasma PI/C ratios, demonstrating intra-islet incretin production in metabolically challenged human islet grafts. These data indicate that in failing grafts, alpha cell prohormone processing is likely altered, and incomplete beta cell prohormone processing may be an early indicator of insulin dependency.

Thyroid hormones (THs) play a critical role in the regulation of biological processes, such as growth, metabolism, and development, in various animal species. Prohormone L-thyroxine (T4) is secreted from the thyroid gland and carried to peripheral tissues. T4 is then biotransformed to several metabolites which play different roles, mainly by iodothyronine deiodinases. Determination of deiodinated TH metabolites in key organs such as liver and brain would help to understand tissue-specific TH metabolism and homeostasis. In this study, we thus developed a highly sensitive method for the determination of six THs [T4, 3,5,3'-triodo-L-thyronine (T3), 3,3',5'-triiodo-L-thyronine (rT3), 3,5-diiodo-l-thyronine (3,5-T2), 3,3'-diiodo-l-thyronine (3,3'-T2), and 3-iodo-l-thyronine (3-T1)] in the brain and liver by using stored dog samples. The analytical method consisted of ultrasonic-assisted extraction in acetone acidified with formic acid, cleanup with a EVOLUTE® EXPRESS CX cartridge (reversed-phase combined with strong cation-exchange cartridge), and quantification with liquid chromatography-tandem mass spectrometry. Acceptable accuracy (internal standard-corrected recovery: 80%-120%) and intra- and inter-day precision (coefficient of variation:6% and15%, respectively) (n = 3/ batch, three days) were obtained for both brain and liver samples. In addition, low method detection limits were achieved for both brain (0.013-0.12 ng g

BackgroundBiopurification has been used to disclose an evolutionarily conserved inhibitory reproductive hormone involved in tissue mass determination. A (rat) bioassay-guided physicochemical fractionation using ovine materials yielded via Edman degradation a 14-residue amino acid (aa) sequence. As a 14mer synthetic peptide (EPL001) this displayed antiproliferative and reproduction-modulating activity, while representing only a part of the native polypeptide. Even more unexpectedly, a scrambled-sequence control peptide (EPL030) did likewise.MethodsReproduction has been investigated in the nematode Steinernema siamkayai, using a fermentation system supplemented with different concentrations of exogenous hexapeptides. Peptide structure-activity relationships have also been studied using prostate cancer and other mammalian cells in vitro, with peptides in solution or immobilized, and via the use of mammalian assays in vivo and through molecular modelling.ResultsReproduction increased (x3) in the entomopathogenic nematode Steinernema siamkayai after exposure to one synthetic peptide (IEPVFT), while fecundity was reduced (x0.5) after exposure to another (KLKMNG), both effects being dose-dependent. These hexamers are opposite ends of the synthetic peptide KLKMNGKNIEPVFT (EPL030). Bioactivity is unexpected as EPL030 is a control compound, based on a scrambled sequence of the test peptide MKPLTGKVKEFNNI (EPL001). EPL030 and EPL001 are both bioinformatically obscure, having no convincing matches to aa sequences in the protein databases. EPL001 has antiproliferative effects on human prostate cancer cells and rat bone marrow cells in vitro. Intracerebroventricular infusion of EPL001 in sheep was associated with elevated growth hormone in peripheral blood and reduced prolactin. The highly dissimilar EPL001 and EPL030 nonetheless have the foregoing biological effects in common in mammalian systems, while being divergently pro- and anti-fecundity respectively in the nematode Caenorhabditis elegans. Peptides up to a 20mer have also been shown to inhibit the proliferation of human cancer and other mammalian cells in vitro, with reproductive upregulation demonstrated previously in fish and frogs, as well as nematodes. EPL001 encodes the sheep neuroendocrine prohormone secretogranin II (sSgII), as deduced on the basis of immunoprecipitation using an anti-EPL001 antibody, with bespoke bioinformatics. Six sSgII residues are key to EPL001’s bioactivity : MKPLTGKVKEFNNI. A stereospecific bimodular tri-residue signature is described involving simultaneous accessibility for binding of the side chains of two specific trios of amino acids, MKP & VFN. An evolutionarily conserved receptor is conceptualised having dimeric binding sites, each with ligand-matching bimodular stereocentres. The bioactivity of the 14mer control peptide EPL030 and its hexapeptide progeny is due to the fortuitous assembly of subsets of the novel hormonal motif, MKPVFN, a default reproductive and tissue-building OFF signal.NOTEPlease see the end of the paper for links to ten files of supplementary information and for an independent peer review.KEY POINTSSynthetic hexapeptides have upregulated and downregulated nematode reproduction, following similar work in other nematode species and in fish and frogs.Peptides up to a 20mer have also been shown to inhibit the proliferation of human cancer and other mammalian cells in vitro and influence circulating levels of ovine pituitary hormones in vivo via intracerebroventricular peptide infusion.The amino acid sequence of the 14mer master peptide arose from a (rat) bioassay- guided fractionation using ovine materials, whose aim was to disclose an evolutionarily conserved inhibitory reproductive hormone involved in tissue mass determination.The bioactive peptides mimic the non-contiguous six-residue receptor-binding active face of a polypeptide hormone, which in mammals is a derivative of the secretory vesicle prohormone secretogranin II (SgII), taking the form of a novel, structually complex 70mer dubbed ‘SgII-70’.The ability has been gained to reinforce or block a default reproductive and tissue- building OFF signal, using peptide mimetics.

IntroductionNucleobindin-2 (NUCB2) was identified as a DNA/Ca2+ binding protein with multiple functions in humans. Prohormone convertase-mediated NUCB2 processing produced nesfatin-1 - a biologically active. Nesfatin-1, an 82-amino acid peptide, was extracted from the N-terminus of nucleobindin-2. Recently, it was described as an anorexia peptide related to weight loss, malnutrition, and appetite regulation in type 2 diabetes mellitus patients.Research design and methodsIn this study, we collected samples and divided them into groups of patients with long-term type 2 diabetes and newly diagnosed type 2 diabetes group. Serum nesfatin-1 level and mRNA NUCB2 gene expression level of the groups were analyzed and compared with those of the healthy group.Biometric parameters and biochemical indices were also analyzed to determine the correlation with nesfatin-1 level.ResultsLevels of nesfatin-1 were found to be higher in the newly diagnosed group than in the other groups. Similar results were also reported in the analysis of mRNA NUCB2 gene expression by Realtime-PCR. Meanwhile, no significant difference was found in both analyzes of nesfatin-1 levels and NUCB2 mRNA expression in subjects with long-term type 2 diabetes compared with the control group. This result can be explained by the effects of long-term treatment. In the correlation of anthropometric parameters and biochemical indices, nesfatin-1 exhibited a significant correlation with BMI (r=0.569), HbA1c (r=-0.468), HDL-C (r=0.731), LDL-C (r=-0.482), Creatinine serum (r=0.525), and Creatinine urine (r=0.592), with pConclusionsThese results indicate that the serum nesfatin-1 level and the NUCB2 mRNA gene expression level may be associated with newly diagnosed type 2 diabetes in Vietnamese patients. However, more specific studies with larger sample sizes were still needed in future studies.

Peptide hormones are first produced as larger precursor prohormones that require endoproteolytic cleavage to liberate the mature hormones. A structurally conserved but functionally distinct family of nine prohormone convertase enzymes (PCs) are responsible for cleavage of protein precursors, of which PC1/3 and PC2 are known to be exclusive to neuroendocrine cells and responsible for prohormone cleavage. Differential expression of PCs within tissues defines prohormone processing; whereas glucagon is the major product liberated from proglucagon via PC2 in pancreatic α-cells, proglucagon is preferentially processed by PC1/3 in intestinal L cells to produce glucagon-like peptides 1 and 2 (GLP-1, GLP-2). Beyond our understanding of processing of islet prohormones in healthy islets, there is convincing evidence that proinsulin, pro-islet amyloid polypeptide (proIAPP), and proglucagon processing is altered during prediabetes and diabetes. There is predictive value of elevated circulating proinsulin or proinsulin-to-C-peptide ratio for progression to type 2 diabetes, and elevated proinsulin or proinsulin-to-C-peptide ratio is predictive for development of type 1 diabetes in at-risk groups. After onset of diabetes, patients have elevated circulating proinsulin and proIAPP, and proinsulin may be an autoantigen in type 1 diabetes. Furthermore, preclinical studies reveal that α-cells have altered proglucagon processing during diabetes, leading to increased GLP-1 production. We conclude that despite strong associative data, current evidence is inconclusive on the potential causal role of impaired prohormone processing in diabetes and suggest that future work should focus on resolving the question of whether altered prohormone processing is a causal driver or merely a consequence of diabetes pathology.

The thyroid hormone system is a main target of endocrine disruptor compounds (EDC) at all levels of its intricately fine-tuned feedback regulation, synthesis, distribution, metabolism and action of the 'prohormone' thyroxine and its active metabolites. Apart from classical antithyroid effects of EDC on the gland, the majority of known and suspected effects occurs at the pre-receptor control of T3 ligand availability to T3 receptors exerting ligand modulated thyroid hormone action. Tissue-, organ- and cell-specific expression and function of thyroid hormone transporters, deiodinases, metabolizing enzymes and T3-receptor forms, all integral components of the system, may mediate adverse EDC effects. Established evidence from nutritional, pharmacological and molecular genetic studies clearly support the functional, biological, and clinical relevance of these targets. Iodine-containing thyroid hormones and the organization of this system are highly conserved during evolution from primitive aquatic life forms, amphibia, birds throughout all vertebrates including humans. Mechanistic studies from various animal experimental models strongly support cause-effect relationships upon EDC exposure, hazards and adverse effects of EDC across various species. Retrospective case-control, cohort and population studies linking EDC exposure with epidemiological data on thyroid hormone-related (dys-)functions provide clear evidence that human development, especially of the fetal and neonatal brain, growth, differentiation and metabolic processes in adult and aging humans are at risk for adverse EDC effects. Considering that more than half of the world population still lives on inadequate iodine supply, the additional ubiquitous exposure to EDC and their mixtures is an additional threat for the essential thyroid hormone system, the health of the human population and their future progenies, animal life forms and our global environment.

Funding Acknowledgements Type of funding sources: None. Introduction Pulmonary arterial hypertension (PAH) is a rare disease, with an incidence of 15–50 cases per million annually. The disease is associated with further progress and worsening of symptoms despite pharmacotherapy. The severity of PAH symptoms and their effects reduce the patients" quality of life (QoL). Numerous studies show that the quality of life is related to the therapeutic effect of the treatment. Available studies demonstrate that patients QoL deteriorates as the condition progresses. Consequently, the goals of PAH therapy have expanded from increasing survival to improving health-related quality of life. The aim of the study was to determine the clinical and sociodemographic factors influencing the quality of life in PAH patients. Methods The study involved 55 patients with PAH who were examined using the Polish version of the PAH-SYMPACT quality of life questionnaire. The PAH-SYMPACT questionnaire results were correlated with several variables. Results It was shown that the quality of life correlates significantly (p˂0,05) with WHO Functional Class, NT-pro BNP (N-terminal prohormone of brain natriuretic peptide) value, elderly age (the higher the value, the worse the QoL). On the other hand, the lower the 6MWT (6 minuts walk test) result, the worse the QoL. Moreover, the intensity of the accompanying PAH symptoms, such as chest pain, dyspnoea, arrhythmias, fatigue and weakness were associated with a lower quality of life in patients (p Conclusion Patients" quality of life correlates with many variables, both clinical and sociodemographic. In addition, there is a correlation between the quality of life of patients with PAH and the severity of several symptoms associated with this condition. Determination of factors affecting the quality of life of patients with PAH enables the selection of a more comprehensive therapy.

Proinsulin, a prohormone precursor to insulin, is synthesized and processed before secretion from pancreatic islet beta cells. Increased plasma proinsulin levels relative to insulin levels may predict insulin resistance and type 2 diabetes (T2D), and the genes involved in proinsulin processing are incompletely understood. Previous genome-wide association studies (GWAS) have identified 13 loci associated with proinsulin levels. To identify additional loci, we conducted a meta-analysis of GWAS results of fasting proinsulin from 16 European-ancestry studies of 45,826 individuals. Each cohort analysis controlled for age, sex, population structure, fasting insulin, and BMI. We identified 35 independent association signals at 30 loci (1 Mb, p Disclosure K. Broadaway: None. X. Yin: None. A. Williamson: None. E. Wilson: None. Magic investigators: n/a.

The objectives of the study were to formulate stable oil-in-water microemulsion (ME) system of cholecalciferol, a lipophilic and unstable prohormone for bone health, and evaluate its pharmacokineti...

Prognostic value of N-terminal fragment of the prohormone brain-type natriuretic peptide (NT-proBNP) was analyzed in patients with multiple myeloma complicated by dialysisdependent renal failure. The prospective study included 20 patients with newly diagnosed multiple myeloma. The concentrations of NT-proBNP were measured before antimyeloma chemotherapy. The median age of the patients was 67 (63-76) years. The median glomerular filtration rate was 4 (4, 5) ml/min/1.73 m2. For overall survival, the area under ROC curve was 0.75 and the cut-off point was 7000 pg/ml. At median follow-up of 17.3 months, the overall survival was 76.6±14.8 and 27.3±13.4% (p=0.02) for cases with NT-proBNP levels below and above the cut-off point, respectively. There were no cases of death due to cardiovascular causes. We concluded that the increase in serum concentration of NT-proBNP>7400 pg/ml is associated with the severity of kidney damage and the risk of non-cardiac mortality.

To closely mimic physiological conditions, low oxygen cultures have been employed in stem cell and cancer research. Although in vivo oxygen concentrations in tissues are often much lower than ambient 21% O2 (ranging from 3.6 to 12.8% O2), most cell cultures are maintained at 21% O2. To clarify the effects of the O2 culture concentration on the regulated secretion of peptide hormones in neuro-endocrine cells, we examined the changes in the storage and release of peptide hormones in neuro-endocrine cell lines and endocrine tissues cultured in a relatively lower O2 concentration. In both AtT-20 cells derived from the mouse anterior pituitary and freshly prepared mouse pituitaries cultured in 10% O2 for 24 h, the storage and regulated secretion of the mature peptide hormone adrenocorticotropic hormone were significantly increased compared with those in cells and pituitaries cultured in ambient 21% O2, whereas its precursor proopiomelanocortin was not increased in the cells and tissues after being cultured in 10% O2. Simultaneously, the prohormone-processing enzymes PC1/3 and carboxypeptidase E were up-regulated in cells cultured in 10% O2, thus facilitating the conversion of prohormones to their active form. Similarly, culturing the mouse β-cell line MIN6 and islet tissue in 10% O2 also significantly increased the conversion of proinsulin into mature insulin, which was secreted in a regulated manner. These results suggest that culture under 10% O2 is more optimal for endocrine tissues/cells to efficiently generate and secrete active peptide hormones than ambient 21% O2.

Vitamin D is an important prohormone largely derived from sunlight exposure on the skin. In many older people, vitamin D levels have been found to be insufficient or frankly deficient. Alterations in body composition associated with ageing, such as decreased muscle mass and increased adipose tissue, may also decrease vitamin D levels. Low vitamin D levels are associated with an increased risk of falls and fractures and the conditions of frailty and sarcopenia, but confounding with decreased outdoor exposure from a large range of concomitant diseases partly explains these findings. The benefits of vitamin D supplementation found in randomised trials have largely been confined to frail and disabled older people. There is some evidence that bolus vitamin D supplements that achieve supraphysiological levels may increase the risk of falls.

Background and Purpose— Recent findings suggest that vitamin D, a neuroprotective prohormone, is involved in the pathogenesis of cardiovascular disease. However, previous studies investigating the association between vitamin D and stroke have shown inconsistent findings. In view of these discrepancies, we determined the association of vitamin D status with stroke using data from a population-based study. Methods— Within the RS (Rotterdam Study), an ongoing prospective population-based study, we measured serum 25-hydroxyvitamin D concentrations between 1997 and 2008 in 9680 participants (56.8% women) aged ≥45 years. We assessed a history of stroke at baseline and subsequently followed for incident stroke until January 1, 2016. Regression models were used to investigate the association of serum 25-hydroxyvitamin D with prevalent and incident stroke separately, adjusted for age, sex, study cohort, season of blood sampling, and other cardiovascular risk factors. Results— Of 9680 participants, 339 had a history of stroke at baseline. Serum 25-hydroxyvitamin D concentration was associated with prevalent stroke, adjusted odds ratio per SD decrease, 1.31; 95% CI, 1.14–1.51. After excluding participants with prevalent stroke, we followed 9338 participants for a total of 98 529 person-years. During follow-up, 735 participants developed a stroke. Lower serum 25-hydroxyvitamin D concentration was not associated with a higher stroke risk, adjusted hazard ratio per SD decrease, 1.06; 95% CI, 0.97–1.16. However, severe vitamin D deficiency did show a significant association: hazard ratio, 1.25; 95% CI, 1.05–1.50. Conclusions— In this population-based cohort, we found an association between vitamin D and prevalent stroke. Only severe vitamin D deficiency was associated with incident stroke. This suggests that lower vitamin D levels do not lead to a higher stroke risk but are instead a consequence of stroke.

Background: Proinsulinoma is a neuroendocrine tumor (NET) of the pancreas that produces prohormone proinsulin. These tumors are very rare. In the literature, they are most often presented in the form of case reports. Materials and Methods: We studied 177 patients with NET of the pancreas who underwent surgical treatment in the A.V. Vishnevsky National Medical Research Centre of Surgery from January 2007 to December 2018. Results: Of 81 patients with organic hyperinsulinism caused by functioning NETs of the pancreas during the study period, 3 (3.7%) had a proinsulinoma; 2 were female; and 1 was male. None of them admitted to weight gain during this period, and their BMI was normal. All patients presented with Whipple’s triad during the 72-h fast. Tumor-enucleating surgery was performed: one robot assisted, two laparotomies. A normal glucose level after treatment was achieved in all cases. Conclusion: In cases where clinical hypoglycemia is present, but the serum insulin level is within the normal range or even decreased, proinsulinoma should be suspected. For now, surgical resection remains the only effective method of treatment. Further investigation of pro­insulinomas is needed.

PubMedID: 30257800 Introduction: A clinical course ranging from mild local findings to life-threatening systemic findings may occur after scorpion stings. The purpose of this study was to identify priority markers indicating scorpion sting–related cardiac involvement. Methods: Our study was performed between July 2014, and September 2015 in the Çukurova University medical faculty pediatric emergency department, in Adana, Turkey. Patients admitted with scorpion sting–related cardiac involvement and a control group consisting of patients with no scorpion sting–related cardiac involvement were included in the study. Troponin I at time of presentation and at 6 and 24 h, N-terminal prohormone of brain natriuretic peptide (NTproBNP), ejection fraction as determined by echocardiography at 24 h, and peak and end of T wave (Tp-e) and Tp-e/QTc ratios with echocardiography at 24 h were evaluated. Results: A patient group consisting of 7 cases of scorpion envenomation–related myocarditis and a control group of 30 cases of scorpion intoxication without myocarditis findings were enrolled. Statistically significantly high glucose, white blood cell values, creatine kinase MB, troponin I, and NTproBNP values were identified in the scorpion sting–related myocarditis group (P

Background About 50% of Americans and 70% of US military service members use dietary supplements (DSs). Objectives This cross-sectional survey examined current prevalence of and factors associated with DS use in service members. Methods A stratified random sample of 200,000 service members from the Air Force, Army, Marine Corps, and Navy was obtained from military manpower records, and these service members were asked to complete a questionnaire on their DS use and personal characteristics. Chi-square statistics and multivariable logistic regression examined differences across various strata of demographic, lifestyle, and military characteristics. Results About 18% of successfully contacted service members (n = 26,681) completed the questionnaire between December 2018 and August 2019 (mean ± SD age: 33 ± 8 y, 86% male). Overall, 74% reported using ≥1 DS/wk. Multivitamins/multiminerals were the most commonly used DSs (45%), followed by combination products (44%), proteins/amino acids (42%), individual vitamins/minerals (31%), herbals (20%), joint health products (9%), and purported prohormones (5%). In multivariable analysis, factors independently associated with DS use included female gender [OR (female/male): 1.91; 95% CI: 1.73, 2.11], older age [OR (≥40/18–24 y): 1.25; 95% CI: 1.08, 1.44], higher education level [OR (college degree/high school or less): 1.35; 95% CI: 1.19, 1.53], higher BMI [OR (≥30/300/≤45 min/wk): 5.05; 95% CI: 4.55, 5.61], smokeless tobacco use [OR (user/nonuser): 1.30; 95% CI: 1.17, 1.44], higher alcohol intake [OR (≥72/0 mL/wk): 1.41; 95% CI: 1.29, 1.54], and higher military rank [OR (senior officer/junior enlisted): 1.26; 95% CI: 1.06, 1.51]. Conclusions Compared with civilian data from the NHANES, service members were much more likely to use DSs and used different types of DSs, especially combination products and proteins/amino acids often used to purportedly enhance physical performance. Comparisons with previous military data suggest DS use has increased over time.

The importance of canonical versus noncanonical mechanisms for the generation of angiotensins remains a major challenge that, in part, is heavily swayed by the relative efficacy of therapies designed to inhibit renin, ACE (angiotensin-converting enzyme), or the Ang II (Angiotensin II) receptor. Ang (1–12) (angiotensin [1-12]) is an Ang II forming substrate serving as a source for Ang II–mediated tissue actions. This study identifies for the first time the presence of Ang (1–12) in the blood of 52 normal (22 women) and 19 (13 women) patients with hypertension not receiving antihypertensive medication at the time of the study. Normal subjects of comparable ages and body habitus had similar circulating plasma Ang (1–12) concentrations (women: 2.02±0.62 [SD] ng/mL; men 2.05±0.55 [SD] ng/mL, P >0.05). The higher values of plasma Ang (1–12) concentrations in hypertensive men (2.51±0.49 ng/mL, n=6) and women (2.33±0.63 [SD] ng/mL, n=13) were statistically significant ( P

Hypertension is associated with inflammation and excessive production of catecholamines. Hypertensive patients have reduced plasma levels of CST (catestatin)—a bioactive cleavage product of the prohormone CgA (chromogranin A). In mouse models, hypertension symptoms can be reduced by administration of CST, but the role of CST in the regulation of cardiovascular function is unknown. In this study, we generated mice with KO (knockout) of the region of the CgA gene coding for CST (CST-KO) and found that CST-KO mice are not only hypertensive as predicted but also display left ventricular hypertrophy, have marked macrophage infiltration of the heart and adrenal gland, and have elevated levels of proinflammatory cytokines and catecholamines. Intraperitoneal injection with CST reversed these phenotypes, and ischemic preconditioning-induced cardioprotection was also abolished in CST-KO mice. Experiments with chlodronate depletion of macrophages and bone marrow transfer showed that macrophages produce CST and that the antihypertensive effects of CST are mediated, in part, via CST’s immunosuppression of macrophages as a form of feedback inhibition. The data thus implicate CST as a key autocrine attenuator of the cardiac inflammation in hypertension by reducing macrophage inflammation.

Background Quantifying the activity of the adrenomedullin system might help to monitor and guide treatment in acute heart failure (AHF) patients. The aims were to (1) identify AHF patients with marked benefit or harm from specific treatments at hospital discharge and (2) predict mortality by quantifying the adrenomedullin system activity. Methods This was a prospective multicentre study. AHF diagnosis and phenotype were centrally adjudicated by two independent cardiologists among patients presenting to the emergency department with acute dyspnoea. Adrenomedullin system activity was quantified using the biologically active component, bioactive adrenomedullin (bio-ADM), and a prohormone fragment, midregional proadrenomedullin (MR-proADM). Bio-ADM and MR-proADM concentrations were measured in a blinded fashion at presentation and at discharge. Interaction with specific treatments at discharge and the utility of these biomarkers on predicting outcomes during 365-day follow-up were assessed. Results Among 1886 patients with adjudicated AHF, 514 patients (27.3%) died during 365-day follow-up. After adjusting for age, creatinine, and treatment at discharge, patients with bio-ADM plasma concentrations above the median (> 44.6 pg/mL) derived disproportional benefit if treated with diuretics (interaction p values n = 764) (interaction p values p p Conclusions Quantifying the adrenomedullin’s system activity may help to personalise post-discharge diuretic treatment and enable accurate risk-prediction in AHF.

Diabetes insipidus (DI) is a syndrome characterized by the persistent excretion of abnormally large volumes of dilute urine. It can be caused by any of four fundamentally different abnormalities: deficient production of the antidiuretic hormone, arginine vasopressin (AVP) by magnocellular neurons that form the posterior pituitary (hypothalamic DI); impaired renal effects of AVP (nephrogenic DI); reduced AVP secretion due to excessive water intake (primary polydipsia); or degradation of AVP by placental vasopressinase (gestational DI). Each type of DI can be caused or potentiated by other disorders. Hypothalamic and nephrogenic DI can also be caused by mutation of the gene that encodes the AVP prohormone, the AVP-2 receptors in the kidney, or the aquaporin-2 water channels that mediate antidiuresis. Familial hypothalamic DI is usually transmitted in an autosomal dominant mode, but autosomal recessive or X-linked recessive forms also exist. Familial nephrogenic DI is usually transmitted in an X-linked recessive mode but can also be autosomal recessive or dominant. Hence the mode of inheritance does not always indicate the type of DI. Indirect methods of differential diagnosis are also unreliable and the pituitary MRI signal is diminished in both types of familial DI. Thus the determination of plasma AVP and/or the response to desmopressin therapy plus gene sequencing provides the best basis for effective management and family counseling.

Background Global longitudinal strain (GLS), most commonly measured at the endocardium, has been shown to be superior to left ventricular (LV) ejection fraction (LVEF) for the identification of systolic dysfunction and prediction of outcomes in heart failure (HF). We hypothesized that strains measured at different myocardial layers (endocardium = ENDO, epicardium = EPI, average = AVE) will have distinct diagnostic and predictive performance for patients with HF. Methods Layer-specific GLS, layer-specific global circumferential strain (GCS) and global radial strain (GRS) were evaluated by cardiovascular magnetic resonance imaging (CMR) feature tracking in the Alberta HEART study. A total of 453 subjects consisted of healthy controls (controls, n = 77), at-risk for HF (at-risk, n = 143), HF with preserved ejection fraction (HFpEF, n = 87), HF with mid-range ejection fraction (HFmrEF, n = 88) and HF with reduced ejection fraction (HFrEF, n = 58). For outcomes analysis, CMR-derived imaging parameters were adjusted with a base model that included age and N-terminal prohormone of b-type natriuretic peptide (NT-proBNP) to test their independent association with 5-year all-cause mortality. Results GLS_EPI distinguished all groups with preserved LVEF (controls − 16.5 ± 2.4% vs. at-risk − 15.5 ± 2.7% vs. HFpEF − 14.1 ± 3.0%, p Conclusion Global strains measured on endocardium, epicardium or averaged across the wall thickness are not equivalent for the identification of systolic dysfunction or outcomes prediction in HF. The endocardium-specific strains were shown to have poorest all-around performance. GLS_AVE and GRS were the only CMR parameters to be significantly associated with 5-year all-cause mortality in multivariable analysis. GLS_EPI and GRS, as well as the difference in endocardial and epicardial strains, were sensitive to systolic dysfunction among HF patients with normal LVEF (> 55%), in whom lower strains were associated with increased concentricity.

The impact of evolution and domestication processes on the sequences of neuropeptide prohormone genes that participate in cell–cell signaling influences multiple biological process that involve neuropeptide signaling. This information is important to understand the physiological differences between Cetartiodactyla domesticated species such as cow, pig, and llama and wild species such as hippopotamus, giraffes, and whales. Systematic analysis of changes associated with evolutionary and domestication forces in neuropeptide prohormone protein sequences that are processed into neuropeptides was undertaken. The genomes from 118 Cetartiodactyla genomes representing 22 families were mined for 98 neuropeptide prohormone genes. Compared to other Cetartiodactyla suborders, Ruminantia preserved PYY2 and lost RLN1. Changes in GNRH2, IAPP, INSL6, POMC, PRLH, and TAC4 protein sequences could result in the loss of some bioactive neuropeptides in some families. An evolutionary model suggested that most neuropeptide prohormone genes disfavor sequence changes that incorporate large and hydrophobic amino acids. A compelling finding was that differences between domestic and wild species are associated with the molecular system underlying ‘fight or flight’ responses. Overall, the results demonstrate the importance of simultaneously comparing the neuropeptide prohormone gene complement from close and distant-related species. These findings broaden the foundation for empirical studies about the function of the neuropeptidome associated with health, behavior, and food production.

Analysis of data from clinical cohorts, and more recently from human pancreatic tissue, indicates that reduced prohormone processing is an early and persistent finding in type 1 diabetes. In this article, we review the current state of knowledge regarding alterations in islet prohormone expression and processing in type 1 diabetes and consider the clinical impact of these findings. Lingering questions, including pathologic etiologies and consequences of altered prohormone expression and secretion in type 1 diabetes, and the natural history of circulating prohormone production in health and disease, are considered. Finally, key next steps required to move forward in this area are outlined, including longitudinal testing of relevant clinical populations, studies that probe the genetics of altered prohormone processing, the need for combined functional and histologic testing of human pancreatic tissues, continued interrogation of the intersection between prohormone processing and autoimmunity, and optimal approaches for analysis. Successful resolution of these questions may offer the potential to use altered prohormone processing as a biomarker to inform therapeutic strategies aimed at personalized intervention during the natural history of type 1 diabetes and as a pathogenic anchor for identification of potential disease-specific endotypes.

Kidney injury is a dangerous diabetic microvascular complication responsible for the mortality of diabetics, which would impose the requirement of biomarkers for early detection of diabetic nephropathy. Copeptin, the C-terminal portion of vasopressin prohormone, is rapidly released in severe endogenous stress. So, the current work was carried out to evaluate and estimate copeptin as a marker for diabetic nephropathy. This cross-sectional work was conducted on 60 female and male type II diabetic cases. Diabetic cases were divided as normoalbuminuric (urinary albumin was 300 mg/24 h) with nephropathy. Control group was designated from fifteen matched healthy subjects. Controls and patients were evaluated for fasting blood glucose, glycosylated hemoglobin (HbA1c), urinary albumin, serum creatinine and serum copeptin. Copeptin concentrations were significantly increased in type II diabetics with macroalbuminuria comparing to healthy controls and diabetics with normoalbuminuria. Serum copeptin concentrations were correlated positively with gold standard urinary albumin, serum creatinine, and HbA1c. Higher serum copeptin concentration in type II diabetics particularly in diabetics with nephropathy and its correlation with urinary albumin and HBA1c reflect the potential role of copeptin as a predictor of diabetes mellitus and development of diabetic nephropathy among type II diabetics considering other risk factors.

Objective Dehydroepiandrosterone sulfate (DHEAs), a prohormone secreted by the adrenal gland, plays a role in the synthesis of sex hormones, namely, androgen and estrogen. It has been found that the amount of DHEAs is correlated with age, although most studies have focused on the correlation of serum DHEAs levels with age and sex. Thus, this noninvasive, cross-sectional study aimed to investigate the correlation of urine DHEAs levels with age and sex in healthy Thai volunteers aged 20–80 years. Methods DHEAs levels were measured in 178 healthy volunteers using electrochemiluminescence immunoassay and then normalized by creatinine. Multiple regression was performed to determine the correlation of urine DHEAs levels normalized by creatinine with age and sex. Results The normalized DHEAs levels are correlated with age group for both sexes. Moreover, an increasing trend in DHEAs levels was found in the age group 20–29 years, and the DHEAs level peaked at the age group 30–39 years before declining with advancing age. Based on the multiple regression analyses, the significance of the interaction term (P ​, Highlights • There is no data on the correlation of urine DHEAs with age and sex in a wide age range. • First report of urine DHEAs levels in healthy Thai volunteers aged 20–80 years. • The fitted model is proposed to determine the correlation of urine DHEAs levels normalized by creatinine with age and sex.

An array of signaling peptides and peptide hormones in pancreatic islets results from posttranslational processing of precursor proteins by well-characterized endopeptidases. While the encoded peptide complement can be predicted in silico from the prohormone sequence, actual peptide products often are more diverse due to additional chemical or enzymatic modifications of mature predicted peptides. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for comprehensive molecular characterization, we compiled an experimentally deduced database of endogenous peptides in human pancreatic islets and revealed an unexpected complex processing of several prohormones including proglucagon. Live pancreatic islets from several donors were provided by the Human Pancreas Analysis Program, University of Pennsylvania. Total of 2034 chemically unique peptide sequences were identified and matched to 477 proteins in the human protein database. Among interesting observations, we find differential cleavage of the signal peptide on the glucagon precursor protein, NP_002045.1, and unusual further processing of its encoded mature peptides. Alternate signal peptide cleavage sites are confirmed by detection of both classical GRPP, R[21]-D[50], and its novel variant S[18]-D[50]. An amidated IP-2 D[131]-L[142] and an isoform of MPGF D[131]-D[178], missing the GLP-1 sequence were detected. Previously unreported N-terminus acetylation of glucagon, GLP-1, GLP-1 (7-36), and oxyntomodulin was detected. Novel peptides potentially resulted from variable processing of mature glucagon, GRPP, and oxyntomodulin at internal convention cleavage sites were characterized. New chemically unique peptides originating from proglucagon and documented in this study indicate a more complex posttranslational processing of proglucagon in human islets than previously thought. The finding expands opportunity for mechanistic studies on pathology, progression, and treatments of diabetes. Disclosure E.V. Romanova: None. S. Rubakhin: None. J.V. Sweedler: None. Funding American Diabetes Association/Pathway to Stop Diabetes (1-18-VSN-19 to J.V.S.)

Proinsulin (PI) is processed to mature insulin by prohormone convertases 1/3 (Pcsk1) and 2 (Pcsk2). In individuals with T1D and T2D, there is an increase in the circulating PI:insulin ratio. We hypothesized that beta-cell Pcsk1 deletion would result in beta-cell dysfunction and hyperglycemia. Male and female Pcsk1flox/flox Ins1cre/+ (Pcsk1beta-KO) and Pcsk1+/+ Ins1cre/+ (Pcsk1beta-WT) mice were fed either a chow or a high fat diet (HFD; 45% fat) and monitored until 30 weeks of age. Despite an approximately 150-fold increase in fasting plasma PI (p < 0.001) and only immature insulin granules detectable by electron microscopy, chow-fed Pcsk1beta-KO male mice had normal glucose tolerance early in life and only developed glucose intolerance by 26 weeks of age (AUC: 934±294 vs. 597±223; p = 0.008). Histological analysis showed that male Pcsk1beta-KO mice displayed a trend toward increased beta-cell area (1.3±0.6 vs. 0.8±0.4%; p = 0.08). The unexpectedly mild hyperglycemia in Pcsk1beta-KO mice is likely due to incomplete disruption of proinsulin processing, as mass spectrometry and western blot analysis of Pcsk1beta-KO islets confirmed markedly elevated proinsulin, but detectable mature insulin. In HFD-fed cohorts, 3/10 Pcsk1beta-KO male mice developed sustained fasting hyperglycemia (>16 mM) after 19 weeks of diet. Chow and HFD-fed female Pcsk1beta-KO mice displayed no differences in glycemia or body weight from Pcsk1beta-WT controls. Deletion of both Pcsk1 and Pcsk2 in beta cells (Pcsk1flox/flox Pcsk2flox/flox Ins1cre/+) resulted in undetectable mature insulin in islets (western blot), and impaired glucose tolerance at 10 weeks of age in both chow-fed male and female mice. Our results suggest that beta-cell Pcsk1 deficiency is not sufficient to drive diabetes development, but increases diabetes susceptibility in conditions of increased insulin secretory demand. Our data in mice support the idea that beta-cell prohormone processing errors likely contribute to pathogenesis of diabetes but not obesity. Disclosure A. Taylor: None. Y. Chen: None. B. Verchere: Advisory Panel; Self; Integrated Nanotherapeutics Inc., Sirona Biochem. Stock/Shareholder; Self; Integrated Nanotherapeutics Inc. Funding Canadian Institutes of Health Research (PJT153156)

Background: Peptide hormones are crucial regulators of metabolism. However, maturation of hormone precursors (prohormones) in the endoplasmic reticulum (ER) in vivo remains largely unexplored. Aims: This project aims to study the integrative role of the two major protein quality control pathways in the ER, ER-Associated Degradation (ERAD) and autophagy, in regulating prohormone maturation under physiological conditions. Taking vasopressin precursor (proAVP) as a model, this project will provide novel insights on hormone maturation and the physiological function of ERQC pathways. Methods: We generated AVP neuron specific ERAD and Autophagy deficient mouse models by deleting Sel1L and Atg7, respectively. We also generated Sel1L and Atg7 deficient cell models. We used immunostaining, sucrose sedimentation, and Western blots to dissect the roles of ERAD and autophagy in proAVP folding and maturation. Results: Mice with Atg7 deficiency in AVP neurons maintain normal water homeostasis, while Sel1L deficient mice develop central diabetes insipidus. Further, Atg7 deficient neurons show reduced proAVP protein and altered subcellular distribution, but its ER export remains functional. Mechanically, unlike Sel1L deficiency, Atg7 deficiency alone shows amber accumulation of misfolded proAVP in vitro; however, further blocking autophagy with ERAD deficiency shows increased portion of misfolded proAVP comparing with ERAD deficiency along. Further, combined deletion of Atg7 and Sel1L in AVP neurons leads to earlier development of Diabetes Insipidus comparing with Sel1L deficiency alone, suggesting a critical role of autophagy in maintaining AVP maturation with compromised ERAD. Conclusions: Here we demonstrate that autophagy alone is dispensable for the maturation of proAVP in the ER, but is indispensable in the event of compromised ERAD function both in vivo and in vitro. This study points to the differential roles and crosstalk between ERAD and autophagy in neuroendocrine cells. Disclosure G. Shi: None. X. Pan: None. H. Wang: None. P. Arvan: None. L. Qi: None. Funding National Institutes of Health (R01DK120047); University of Michigan (U064133)

There is no satisfactory answer on the specific biomarker that might be used in differentiating heart failure with reduced EF (HFrEF), allowing for inadequacy of N-terminal prohormone brain natriuretic peptide (NT-proBNP).We aim to evaluate the value of microRNA-208a in diagnosing HFrEF patients.We included 120 HF patients and 60 healthy volunteers. Diagnostic values of NT-proBNP and miR-208a for HF patients versus controls and HFrEF versus HFpEF were described by area under curve (AUC), sensitivity and specificity.HFrEF patients had significantly higher miR-208a level (p0.001). As for diagnosing HFrEF patients, additional use of miR-208a and NT-proBNP yielded a significantly higher AUC than NT-proBNP alone (0.83, 95% CI 0.76-0.90 vs. 0.73, 95% CI 0.64-0.82) and the sensitivity and specificity were raised to 68.0% and 90.2%.Use of miR-208a in combination with NT-proBNP may allow a more reliable method in diagnosing HFrEF patients.

Hypertension (HTN) is a pandemic associated with inflammation and excessive production of catecholamines. Previous work has shown that hypertensive patients have reduced plasma levels of Catestatin (CST), a bioactive cleavage product of the prohormone Chromogranin A (CgA). Similarly, in mouse models, HTN symptoms can be reduced by administration of CST, but the role of CST in the regulation of cardiovascular function is unknown. In the present study, we generated mice with knockout (KO) of the region of the CgA gene coding for CST (CST-KO) and found that CST-KO mice are not only hypertensive as predicted, but also display left ventricular hypertrophy, have marked macrophage infiltration of the heart and adrenal gland, and have elevated levels of pro-inflammatory cytokines and catecholamines. Additionally, intraperitoneal injection with CST reverses these phenotypes, and ischemic pre-conditioning-induced cardioprotection was also abolished in CST-KO mice. To further explore the relationship between HTN, CST, and macrophages, experiments with chlodronate depletion of macrophages and bone-marrow transfer showed that macrophages produce CST and that the anti-hypertensive effects of CST are mediated in part via immunosuppression of macrophages by CST as a form of feedback inhibition. The data thus implicate CST as a key autocrine attenuator of the cardiac inflammation in HTN by reducing macrophage inflammation.

Hormones transferred from mothers to their offspring are considered a maternal tool to prepare progeny for expected environmental conditions, increasing maternal fitness. To flexibly influence offspring, mothers should be able to transmit the hormonal signals independent of their own hormonal status. However, the ability to regulate hormone transfer to the next generation is under debate. We studied the transfer of thyroid hormones (THs) to eggs in a bird model. We elevated thyroxine (T4, the prohormone for the biological active triiodothyronine, T3) during egg-laying using T4 implants on females of a wild population of pied flycatchers (Ficedula hypoleuca), and measured plasma and yolk T4 and T3 as a response. To our knowledge, studies that manipulated a prohormone and measured the change in its active metabolites have rarely been conducted. We found an increase in plasma and yolk T4 and no change in T3 concentrations leading to a similar decrease in yolk T3/T4 ratio in response to the T4 treatment in plasma and yolk. This suggests that mothers are able to regulate the conversion of T4 in T3 to avoid potential costs of elevated exposure to the active hormone to herself and to her progeny. Finally, contrary to our predictions, we found no evidence of regulatory mechanisms at the follicle level, which is essential for independent regulation of yolk hormone transfer.Summary statementThyroid hormones have been overlooked in the context of hormone-mediated maternal effects. We found that mothers may regulate yolk thyroid hormone transfer by regulating the conversion of the active form of the hormone.

Maternal thyroid hormones (THs) are known to be crucial in embryonic development in humans, but their influence on other, especially wild, animals remains poorly understood. So far, the studies that experimentally investigated the consequences of maternal THs focused on short-term effects, while early organisational effects with long-term consequences, as shown for other prenatal hormones, could also be expected. In this study, we aimed at investigating both the short- and long-term effects of prenatal THs in a bird species, the Japanese quail Coturnix japonica. We experimentally elevated yolk TH content (the prohormone T4, and its active metabolite T3, as well as a combination of both hormones). We analysed hatching success, embryonic development, offspring growth and oxidative stress as well as their potential organisational effects on reproduction, moult and oxidative stress in adulthood. We found that eggs injected with T4 had a higher hatching success compared with control eggs, suggesting conversion of T4 into T3 by the embryo. We detected no evidence for other short-term or long-term effects of yolk THs. These results suggest that yolk THs are important in the embryonic stage of precocial birds, but other short- and long-term consequences remain unclear. Research on maternal THs will greatly benefit from studies investigating how embryos use and respond to this maternal signalling. Long-term studies on prenatal THs in other taxa in the wild are needed for a better understanding of this hormone-mediated maternal pathway.

Background: The diagnosis of diabetes insipidus (DI) relies on indirect measurement of serum and urine sodium and osmolality. Since the diagnosis can only be made when an inappropriately dilute urine is paired with a significantly concentrated serum, the process is tedious for the clinician and uncomfortable for the patient. Copeptin is the C-terminal portion of the anti-diuretic hormone (ADH) prohormone which correlates with the less stable ADH, therefore providing a direct measurement of posterior pituitary response to hyperosmolar stress. (1,2) Aim: This study aims to assess the diagnostic accuracy of copeptin in patients with central DI compared with subjects who underwent pituitary surgery without developing DI. Methods: Serum samples from subjects with central DI, control subjects post pituitary surgery with no DI (NDI) and control subjects with SIADH were collected and analysed on the BRAHMS KRYPTOR copeptin assay. Groups were compared using unpaired T-test and Levene’s test for equal variance. Results: 56 samples from 22 subjects (13 females, 9 males, mean age 53.9 ± 15.5 y.o.) were analysed. Two subjects had resolved DI (RDI) after copeptin analysis and were successfully weaned off DDAVP and reclassified as NDI. Of the DI subjects, 1 had acute and 5 had chronic DI. Copeptin was lower in DI compared to NDI group (p = 0.013), while serum sodium, osmolality, urine osmolality were similar. Copeptin did not differentiate between the SIADH and NDI groups. After exclusion of NDI samples with serum sodium ≤ 140 mmol/L, the area under the curve was 0.97 (95% CI 0.9 to 1.0), a copeptin cut-off of 2.9 pmol/L predicts DI with a sensitivity of 92% and a specificity of 90%. Conclusion: Copeptin concentration of < 3.0 pmol/L concurrently with serum sodium concentration of > 140mmol/L predicted central DI when using post pituitary surgery subjects without DI as controls. 1. Winzeler, B., Zweifel, C., Nigro, N., Arici, B., Bally, M., Schuetz, P., Blum, C., Kelly, C., Berkmann, S., Huber, A., Gentili, F., Zadeh, G., Landolt, H., Mariani, L., Müller, B. and Christ-Crain, M. (2015). Postoperative Copeptin Concentration Predicts Diabetes Insipidus After Pituitary Surgery. The Journal of Clinical Endocrinology & Metabolism, 100(6), pp.2275-2282. 2. Fenske, J., Refardt, I., SchnyderI., Winzeler, B., Drummond J., Ribeiro-Oliveira, Jr. A., Drescher, T., Bilz S., Vogt, D.R., Malzahn, U., Kroiss, M., Christ, E., Henzen, C., Fischli S., Tönjes, A., Mueller, B., Schopohl, J., Flitsch, J., Brabant, G., Fassnacht M., Christ-Crain, M. (2018). Copeptin in the Diagnosis of Diabetes Insipidus. New England Journal of Medicine, 379(18), pp.1784-1786.

Anti-mullerian hormone (AMH) is a cytokine of transforming growth factor β (TGF-β) superfamily able to induce apoptosis in cells bearing specific AMH type II receptors (AMHRII). AMHRII is overexpressed in some malignant cells, so at present recombinant AMH (rAMH) is considered as a new candidate antineoplastic drug. The use of rAMH may be especially effective in case of such severe diseases as ovarian, prostate and breast cancer. However, the development of a new drug is hampered by the laboriousness of obtaining highly purified rAMH and by the lack of data about the pharmacological characteristics of rAMH derivatives. In this work, we obtained preparations of prohormone, half-cleaved rAMH and a C-terminal fragment of rAMH, which was confirmed by qualitative and quantitative analyses. To obtain rAMH and its derivatives we used a previously developed highly effective producer strain containing the optimized human AMH gene. The production process has been divided into several stages: (a) rAMH biosynthesis in the bioreactor; (b) culture media preparation; (c) purification of rAMH and its derivatives using immunoaffinity chromatography and reversed-phase HPLC; (d) identification of the purified proteins by immunoblotting and analytical reversed-phase HPLC; and (e) evaluation of the hormone forms activity. The obtained proteins may be used in preclinical trials and in vitro study of rAMH derivatives properties.