Journal of Physiology-London最新文献

{"title":"Annual Prize Lecture 2024: Endogenous physiological mechanisms as basis for the treatment of obesity and type 2 diabetes.","authors":"Jens Juul Holst","doi":"10.1113/JP287461","DOIUrl":"https://doi.org/10.1113/JP287461","url":null,"abstract":"<p><p>In 1964, it was proven that postprandial insulin secretion is largely regulated by gut hormones and, in 1973, it was proposed that a gut hormone would also regulate appetite and food intake. Several gut hormones were tested for metabolic actions with disappointing results until the discovery of the proglucagon derivative, glucagon-like peptide-1 (GLP-1). This peptide from the distal intestine has preserved activity on insulin secretion in people with type 2 diabetes and turned out to regulate both secretion and motility in the gastrointestinal tract and importantly, appetite and food intake, thus functioning as an efficient 'ileal brake' hormone. However, the natural hormone acts predominantly via sensory afferent systems and is extremely rapidly removed from the circulation by enzymatic degradation and renal elimination, and increasing the doses merely results in nausea and vomiting. Lipidation of analogs turned out to provide both stability and limit renal elimination, and very slow up-titration of dosing improves tolerance. Indeed, the most recent agonists may near-normalize glycaemic control in type 2 diabetes, may cause weight losses of up to 25% of body weight, and significantly reduce cardiovascular risk, effects that resemble those of bariatric surgery. Thus, a solution to one of the most serious health problems of modern civilization, the increased morbidity and mortality of the metabolic syndrome, may be addressed by mobilization of one of the body's own regulatory mechanisms.</p>","PeriodicalId":50088,"journal":{"name":"Journal of Physiology-London","volume":" ","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142631658","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Daily GDF15 treatment has sex-specific effects on body weight and food intake and does not enhance the effects of voluntary physical activity in mice.","authors":"Stewart Jeromson, Michael Akcan, Bradley Baranowski, Meagan Arbeau, Annalaura Bellucci, David C Wright","doi":"10.1113/JP287256","DOIUrl":"https://doi.org/10.1113/JP287256","url":null,"abstract":"<p><p>Growth differentiation factor 15 (GDF15) is a stress-induced cytokine that suppresses food intake and causes weight loss. GDF15 also reduces voluntary physical activity and, thus, it is not clear whether combining GDF15 with exercise will be beneficial or if reductions in food intake would be offset by decreases in physical activity. We investigated how GDF15 treatment combined with voluntary wheel running (VWR) would impact weight gain, food intake, adiposity and indices of metabolic health in mice. High-fat fed male and female mice underwent daily GDF15 treatments and were given access to voluntary running wheels, or not, for 11 days. In both sexes, VWR prevented weight gain. In males, GDF15 reduced food intake, as well as attenuated weight gain and the accumulation of adipose tissue, with no additional effect of VWR. In female mice, GDF15 did not impact body weight gain or body composition. GDF15 acutely reduced food intake in female mice but this was followed by a period of rebound hyperphagia and consequently GDF15 did not reduce total food intake in female mice. GDF15 treatment reduced wheel running distance in both sexes. There were main effects of VWR to improve glucose tolerance in female but not male mice. These findings show that GDF15 has sex-specific effects on food intake and consequently weight gain and adiposity. There is no added benefit of combining GDF15 and voluntary physical activity for weight loss. Adaptive responses to acute caloric restriction induced by GDF15 might limit its effectiveness as a weight loss tool in females. KEY POINTS: GDF15 is a stress-induced signalling factor that reduces food intake and voluntary physical activity. It is not known whether combining GDF15 treatment with voluntary wheel running would impart beneficial combined effects in attenuating weight gain and the accumulation of adipose tissue. In the present study, we demonstrate that GDF15 reduces food intake and prevents weight gain in male but not female mice consuming a high-fat diet and also that combining GDF15 with voluntary wheel running (VWR) does not lead to a greater dampening of weight gain. In female mice, GDF15 acutely reduced food intake, but this was followed by a period of rebound hyperphagia resulting in no differences in total food intake. In both sexes, VWR was equivalent, or superior to GDF15 in preventing weight gain.</p>","PeriodicalId":50088,"journal":{"name":"Journal of Physiology-London","volume":" ","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142631669","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Skeletal muscle reactive oxygen species and microvascular endothelial function in age-related hypertension: a study protocol using a microdialysis technique.","authors":"Hollie Speer, Mostafa M Ali, Nathan M D'Cunha, Nenad Naumovski, Stephan F E Praet, Robert C Hickner, Andrew J McKune","doi":"10.1113/JP287187","DOIUrl":"https://doi.org/10.1113/JP287187","url":null,"abstract":"<p><p>Increased reactive oxygen species (ROS) generation and microvascular endothelial disruptions occur with natural ageing, but often transpire before the detection of cardiometabolic conditions including hypertension. Age-related increases in blood pressure are driven by complex systemic changes with poorly understood integrated mechanisms. The deconditioning experienced by ageing skeletal muscle from mid-life is associated with reduced microvascular blood flow and increased peripheral resistance, suggesting that vasodilatory decrements in the muscle may precede the age-related increases in blood pressure. Structural and functional changes within the vascular and skeletal muscle systems with advancing age can influence redox homeostasis, and vice versa, further compounding microvascular endothelial dysfunction. Therefore, comparisons between the microvascular environments of healthy and hypertensive cohorts can provide insights into the changes that occur during significant periods of functional decline. This comprehensive study protocol describes a microdialysis technique to assess the interactions of microvascular health and functional changes in the muscle, which currently cannot be otherwise addressed. Here, we detail an experimental protocol to simultaneously detect skeletal muscle ROS (H₂O₂ and indirect O₂ ^-), determine nutritive blood flow and assess microvascular endothelial function in response to acetylcholine stimulation. We expect that healthy middle-aged individuals should not have increased ROS generation in the muscle at rest, compared to their hypertensive or older counterparts, but may exhibit perturbed microvascular function. The described technique allows for intricate exploration of microvascular physiology that will provide a critically novel insight into benchmarking potential age-related mechanisms involved in the development of age-related hypertension, and aid in early identification and prevention. KEY POINTS: Increased reactive oxygen species (ROS) production and microvascular endothelial dysfunction precede the onset of age-related cardiometabolic and vascular conditions such as hypertension. The profound structural and functional changes that occur within the vasculature and in skeletal muscle from middle age prompt a need to mechanistically explore the microvascular environment in healthy and hypertensive individuals. Using a novel microdialysis technique, we detail an experimental protocol to simultaneously detect skeletal muscle ROS (H₂O₂ and indirect O₂ ^-), determine nutritive blood flow and assess microvascular endothelial function in response to acetylcholine stimulation. With this technique and study protocol, we can reveal functional insights into potential perturbations in ROS generation at rest and the microvascular endothelium, which play important roles in the development of age-related hypertension.</p>","PeriodicalId":50088,"journal":{"name":"Journal of Physiology-London","volume":" ","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142631745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Regional cerebral perfusion and sympathetic activation during exercise in hypoxia and hypercapnia: preliminary insight into 'Cushing's mechanism'.","authors":"Hsuan-Yu Wan, Kanokwan Bunsawat, Catherine L Jarrett, Katherine L Shields, Angela V Bisconti, Joshua C Weavil, Markus Amann","doi":"10.1113/JP287181","DOIUrl":"10.1113/JP287181","url":null,"abstract":"&lt;p&gt;&lt;p&gt;We examined the interactive influence of hypoxia and exercise, and hypercapnia and exercise, on regional cerebral perfusion and sympathetic activation. Twenty healthy young adults (seven women) completed study trials including (1) rest in normoxia ( &lt;math&gt; &lt;semantics&gt;&lt;msub&gt;&lt;mi&gt;S&lt;/mi&gt; &lt;mrow&gt;&lt;mi&gt;p&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;O&lt;/mi&gt; &lt;mn&gt;2&lt;/mn&gt;&lt;/msub&gt; &lt;/mrow&gt; &lt;/msub&gt; &lt;annotation&gt;${{S}_{{mathrm{p}}{{{mathrm{O}}}_{mathrm{2}}}}}$&lt;/annotation&gt;&lt;/semantics&gt; &lt;/math&gt; : ∼96%, &lt;math&gt; &lt;semantics&gt;&lt;msub&gt;&lt;mi&gt;P&lt;/mi&gt; &lt;mrow&gt;&lt;mi&gt;ETC&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;O&lt;/mi&gt; &lt;mn&gt;2&lt;/mn&gt;&lt;/msub&gt; &lt;/mrow&gt; &lt;/msub&gt; &lt;annotation&gt;${{P}_{{mathrm{ETC}}{{{mathrm{O}}}_{mathrm{2}}}}}$&lt;/annotation&gt;&lt;/semantics&gt; &lt;/math&gt; : ∼36 mmHg), normocapnic hypoxia ( &lt;math&gt; &lt;semantics&gt;&lt;msub&gt;&lt;mi&gt;S&lt;/mi&gt; &lt;mrow&gt;&lt;mi&gt;p&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;O&lt;/mi&gt; &lt;mn&gt;2&lt;/mn&gt;&lt;/msub&gt; &lt;/mrow&gt; &lt;/msub&gt; &lt;annotation&gt;${{S}_{{mathrm{p}}{{{mathrm{O}}}_{mathrm{2}}}}}$&lt;/annotation&gt;&lt;/semantics&gt; &lt;/math&gt; : ∼84%, &lt;math&gt; &lt;semantics&gt;&lt;msub&gt;&lt;mi&gt;P&lt;/mi&gt; &lt;mrow&gt;&lt;mi&gt;ETC&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;O&lt;/mi&gt; &lt;mn&gt;2&lt;/mn&gt;&lt;/msub&gt; &lt;/mrow&gt; &lt;/msub&gt; &lt;annotation&gt;${{P}_{{mathrm{ETC}}{{{mathrm{O}}}_{mathrm{2}}}}}$&lt;/annotation&gt;&lt;/semantics&gt; &lt;/math&gt; : ∼36 mmHg), and normoxic hypercapnia ( &lt;math&gt; &lt;semantics&gt;&lt;msub&gt;&lt;mi&gt;S&lt;/mi&gt; &lt;mrow&gt;&lt;mi&gt;p&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;O&lt;/mi&gt; &lt;mn&gt;2&lt;/mn&gt;&lt;/msub&gt; &lt;/mrow&gt; &lt;/msub&gt; &lt;annotation&gt;${{S}_{{mathrm{p}}{{{mathrm{O}}}_{mathrm{2}}}}}$&lt;/annotation&gt;&lt;/semantics&gt; &lt;/math&gt; : ∼98%, &lt;math&gt; &lt;semantics&gt;&lt;msub&gt;&lt;mi&gt;P&lt;/mi&gt; &lt;mrow&gt;&lt;mi&gt;ETC&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;O&lt;/mi&gt; &lt;mn&gt;2&lt;/mn&gt;&lt;/msub&gt; &lt;/mrow&gt; &lt;/msub&gt; &lt;annotation&gt;${{P}_{{mathrm{ETC}}{{{mathrm{O}}}_{mathrm{2}}}}}$&lt;/annotation&gt;&lt;/semantics&gt; &lt;/math&gt; : ∼46 mmHg) and (2) unilateral rhythmic handgrip exercise (45% of maximal voluntary contraction at 1 Hz for 3 min) under the same gas conditions. Based on the exercising arm, blood flow in the contralateral internal carotid (ICA&lt;sub&gt;BF&lt;/sub&gt;) and ipsilateral vertebral (VA&lt;sub&gt;BF&lt;/sub&gt;) arteries, anterior and posterior cerebral O&lt;sub&gt;2&lt;/sub&gt; delivery ( &lt;math&gt; &lt;semantics&gt;&lt;mrow&gt;&lt;mi&gt;C&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;D&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;O&lt;/mi&gt; &lt;mn&gt;2&lt;/mn&gt;&lt;/msub&gt; &lt;/msub&gt; &lt;/mrow&gt; &lt;annotation&gt;${mathrm{C}}{{{mathrm{D}}}_{{{{mathrm{O}}}_{mathrm{2}}}}}$&lt;/annotation&gt;&lt;/semantics&gt; &lt;/math&gt; ), and muscle sympathetic nerve activity (MSNA) were measured in each trial. During exercise in hypoxia, ICA&lt;sub&gt;BF&lt;/sub&gt;, VA&lt;sub&gt;BF&lt;/sub&gt;, anterior and posterior &lt;math&gt; &lt;semantics&gt;&lt;mrow&gt;&lt;mi&gt;C&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;D&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;O&lt;/mi&gt; &lt;mn&gt;2&lt;/mn&gt;&lt;/msub&gt; &lt;/msub&gt; &lt;/mrow&gt; &lt;annotation&gt;${mathrm{C}}{{{mathrm{D}}}_{{{{mathrm{O}}}_{mathrm{2}}}}}$&lt;/annotation&gt;&lt;/semantics&gt; &lt;/math&gt; were significantly lower, whereas total MSNA was significantly greater, than the sum of the responses evoked by either hypoxia or exercise alone. During exercise in hypercapnia, ICA&lt;sub&gt;BF&lt;/sub&gt; and anterior &lt;math&gt; &lt;semantics&gt;&lt;mrow&gt;&lt;mi&gt;C&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;D&lt;/mi&gt; &lt;msub&gt;&lt;mi&gt;O&lt;/mi&gt; &lt;mn&gt;2&lt;/mn&gt;&lt;/msub&gt; &lt;/msub&gt; &lt;/mrow&gt; &lt;annotation&gt;${mathrm{C}}{{{mathrm{D}}}_{{{{mathrm{O}}}_{mathrm{2}}}}}$&lt;/annotation&gt;&lt;/semantics&gt; &lt;/math&gt; were significantly greater, whereas MSNA was lower, than the sum of the responses evoked by either hyperc","PeriodicalId":50088,"journal":{"name":"Journal of Physiology-London","volume":" ","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142631743","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The alarmin, interleukin-33, increases vascular tone via extracellular signal regulated kinase-mediated Ca²⁺ sensitization and endothelial nitric oxide synthase inhibition.","authors":"Evan DeVallance, Elizabeth Bowdridge, Krista Garner, Julie Griffith, Madison Seman, Thomas Batchelor, Murugesan Velayutham, W Travis Goldsmith, Salik Hussain, Eric E Kelley, Timothy R Nurkiewicz","doi":"10.1113/JP286990","DOIUrl":"10.1113/JP286990","url":null,"abstract":"<p><p>Alarmins are classified by their release from damaged or ruptured cells. Many alarmins have been found to increase vascular tone and oppose endothelium-dependent dilatation (EDD). Interleukin (IL)-33 plays a prominent role in lung injury and can be released during vascular injury and in chronic studies found to be cardioprotective. Our recent work has implicated IL-33 in acute vascular dysfunction following inhalation of engineered nanomaterials (ENM). However, the mechanisms linking IL-33 to vascular tone have not been interrogated. We therefore aimed to determine whether IL-33 directly influenced microvascular tone and endothelial function. Isolated feed arteries and in vivo arterioles from male and female Sprague-Dawley rats were used to determine direct vascular actions of IL-33. Mesenteric feed arteries and arterioles demonstrated reduced intraluminal diameters when treated with increasing concentrations of recombinant IL-33. IL-33 activated extracellular signal regulated kinase (ERK)1/2 of rat aortic smooth muscle cells but not phosphorylation of myosin light chain kinase. This suggested IL-33 may sensitize arterioles to Ca²⁺-mediated responses. Indeed, IL-33 augmented the myogenic- and phenylephrine-induced vasoconstriction. Additionally, incubation of arterioles with 1 ng IL-33 attenuated ACh-mediated EDD. Mechanistically, in human aortic endothelial cells, we demonstrate that IL-33-mediated ERK1/2 activation leads to inhibitory phosphorylation of serine 602 on endothelial nitric oxide synthase. Finally, we demonstrate that IL-33-ERK1/2 contributes to vascular tone following two known inducers of IL-33; ENM inhalation and the rupture endothelial cells. The present study provides novel evidence that IL-33 increases vascular tone via canonical ERK1/2 activation in microvascular smooth muscle and endothelium. Altogether, it is suggested IL-33 plays a critical role in microvascular homeostasis following barrier cell injury. KEY POINTS: Interleukin (IL)-33 causes a concentration-dependent reduction in feed artery diameter. IL-33 acts on vascular smooth muscle cells to augment Ca²⁺-mediated processes. IL-33 causes inhibitory phosphorylation of endothelial nitric oxide synthase and opposes endothelium-dependent dilatation. Engineered nanomaterial-induced lung injury and endothelial cell rupture in part act through IL-33 to mediate increased vascular tone.</p>","PeriodicalId":50088,"journal":{"name":"Journal of Physiology-London","volume":" ","pages":"6087-6107"},"PeriodicalIF":4.7,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142629937","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Could cystic fibrosis drugs be repurposed for treating alcohol-induced pancreatitis?","authors":"Pawel E Ferdek","doi":"10.1113/JP287723","DOIUrl":"10.1113/JP287723","url":null,"abstract":"","PeriodicalId":50088,"journal":{"name":"Journal of Physiology-London","volume":" ","pages":"5985-5986"},"PeriodicalIF":4.7,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142631663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impaired oxytocin signalling in the central amygdala in rats with chronic heart failure.","authors":"Ferdinand Althammer, Ranjan K Roy, Matthew K Kirchner, Elba Campos Lira, Stephanie Schimmer, Alexandre Charlet, Valery Grinevich, Javier E Stern","doi":"10.1113/JP286297","DOIUrl":"10.1113/JP286297","url":null,"abstract":"<p><p>Heart failure (HF) patients suffer from cognitive decline and mood impairments, but the molecular signals and brain circuits underlying these effects remain elusive. The hypothalamic neuropeptide oxytocin (OT) is critically involved in regulating mood, and OTergic signalling in the central amygdala (CeA) is a key mechanism that controls emotional responses including anxiety-like behaviours. Still, whether an altered OTergic signalling contributes to mood disorders in HF remains unknown. To address this, we used an ischaemic rat HF model, along with a highly multidisciplinary approach, to mechanistically study multiple levels of the hypothalamus-to-CeA OTergic circuit in male rats with HF. We aimed to test the hypothesis that sustained activation of the OT system following an infarct leads to depletion of OT content in this pathway, with subsequent changes in OT receptor expression and blunted modulation of local GABAergic circuits. We found that most of OTergic innervation of the CeA originated from the supraoptic nucleus (SON). While no differences in the numbers of SON→CeA OTergic neurons was observed between sham and HF rats, we observed a blunted content and release of OT from axonal terminals within the CeA. Moreover, we report downregulation of neuronal and astrocytic OT receptors, and impaired OTR-driven GABAergic synaptic activity within the CeA microcircuit of HF rats. We provide the first evidence that male HF rats display perturbations in the hypothalamus-to-amygdala OTergic circuit, laying the foundation for future translational studies targeting either the OT system or GABAergic amygdalar microcircuit to ameliorate mood impairments in rats or patients with chronic HF. KEY POINTS: Heart failure patients suffer from cognitive decline, depression and mood impairments, but the underlying mechanisms remain elusive. Acting within the central amygdala, the neuropeptide oxytocin regulates emotional responses, including anxiety-like behaviours. However, whether changes in oxytocin signalling occurs during heart failure is unknown. In this study, we used an ischaemic rat heart failure model to mechanistically study multiple levels of the hypothalamus-to-amygdala oxytocinergic circuit in this disease. We report an overall blunted oxytocinergic signalling pathway in rats with heart failure, including blunted content and release of oxytocin from axonal terminals, downregulation of neuronal and astrocytic oxytocin receptors, and impaired oxytocin-driven GABAergic synaptic activity within the central amygdala microcircuit of HF rats. These studies shed light on mechanisms that contribute to mood disorders in cardiovascular disease states and help to identify potential molecular targets for their improved treatment.</p>","PeriodicalId":50088,"journal":{"name":"Journal of Physiology-London","volume":" ","pages":"6259-6280"},"PeriodicalIF":4.7,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142631678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}