CARD-only proteins regulate in vivo inflammasome responses and ameliorate gout.

Inflammatory responses are crucial for controlling infections and initiating tissue repair. However, excessive and uncontrolled inflammation causes inflammatory disease. Processing and release of the pro-inflammatory cytokines interleukin-1ß (IL-1ß) and IL-18 depend on caspase-1 activation within inflammasomes. Assembly of inflammasomes is initiated upon activation of cytosolic pattern recognition receptors (PRRs), followed by sequential polymerization of pyrin domain (PYD)-containing and caspase recruitment domain (CARD)-containing proteins mediated by homotypic PYD and CARD interactions. Small PYD- or CARD-only proteins (POPs and COPs, respectively) evolved in higher primates to target these crucial interactions to limit inflammation. Here, we show the ability of COPs to regulate inflammasome activation by modulating homotypic CARD-CARD interactions in vitro and in vivo. CARD16, CARD17, and CARD18 displace crucial CARD interactions between caspase-1 proteins through competitive binding and ameliorate uric acid crystal-mediated NLRP3 inflammasome activation and inflammatory disease. COPs therefore represent an important family of inflammasome regulators and ameliorate inflammatory disease.

Functional changes of the gastric bypass microbiota reactivate thermogenic adipose tissue and systemic glucose control via intestinal FXR-TGR5 crosstalk in diet-induced obesity.

BACKGROUND: Bariatric surgery remains the most effective therapy for adiposity reduction and remission of type 2 diabetes. Although different bariatric procedures associate with pronounced shifts in the gut microbiota, their functional role in the regulation of energetic and metabolic benefits achieved with the surgery are not clear. METHODS: To evaluate the causal as well as the inherent therapeutic character of the surgery-altered gut microbiome in improved energy and metabolic control in diet-induced obesity, an antibiotic cocktail was used to eliminate the gut microbiota in diet-induced obese rats after gastric bypass surgery, and gastric bypass-shaped gut microbiota was transplanted into obese littermates. Thorough metabolic profiling was combined with omics technologies on samples collected from cecum and plasma to identify adaptions in gut microbiota-host signaling, which control improved energy balance and metabolic profile after surgery. RESULTS: In this study, we first demonstrate that depletion of the gut microbiota largely reversed the beneficial effects of gastric bypass surgery on negative energy balance and improved glucolipid metabolism. Further, we show that the gastric bypass-shaped gut microbiota reduces adiposity in diet-induced obese recipients by re-activating energy expenditure from metabolic active brown adipose tissue. These beneficial effects were linked to improved glucose homeostasis, lipid control, and improved fatty liver disease. Mechanistically, these effects were triggered by modulation of taurine metabolism by the gastric bypass gut microbiota, fostering an increased abundance of intestinal and circulating taurine-conjugated bile acid species. In turn, these bile acids activated gut-restricted FXR and systemic TGR5 signaling to stimulate adaptive thermogenesis. CONCLUSION: Our results establish the role of the gut microbiome in the weight loss and metabolic success of gastric bypass surgery. We here identify a signaling cascade that entails altered bile acid receptor signaling resulting from a collective, hitherto undescribed change in the metabolic activity of a cluster of bacteria, thereby readjusting energy imbalance and metabolic disease in the obese host. These findings strengthen the rationale for microbiota-targeted strategies to improve and refine current therapies of obesity and metabolic syndrome. Video Abstract Bariatric Surgery (i.e. RYGB) or the repeated fecal microbiota transfer (FMT) from RYGB donors into DIO (diet-induced obesity) animals induces shifts in the intestinal microbiome, an effect that can be impaired by oral application of antibiotics (ABx). Our current study shows that RYGB-dependent alterations in the intestinal microbiome result in an increase in the luminal and systemic pool of Taurine-conjugated Bile acids (TCBAs) by various cellular mechanisms acting in the intestine and the liver. TCBAs induce signaling via two different receptors, farnesoid X receptor (FXR, specifically in the intestines) and the G-protein-coupled bile acid receptor TGR5 (systemically), finally resulting in metabolic improvement and advanced weight management. BSH, bile salt hydrolase; BAT brown adipose tissue.

NLRP3 licenses NLRP11 for inflammasome activation in human macrophages.

Intracellular sensing of stress and danger signals initiates inflammatory innate immune responses by triggering inflammasome assembly, caspase-1 activation and pyroptotic cell death as well as the release of interleukin 1ß (IL-1ß), IL-18 and danger signals. NLRP3 broadly senses infectious patterns and sterile danger signals, resulting in the tightly coordinated and regulated assembly of the NLRP3 inflammasome, but the precise mechanisms are incompletely understood. Here, we identified NLRP11 as an essential component of the NLRP3 inflammasome in human macrophages. NLRP11 interacted with NLRP3 and ASC, and deletion of NLRP11 specifically prevented NLRP3 inflammasome activation by preventing inflammasome assembly, NLRP3 and ASC polymerization, caspase-1 activation, pyroptosis and cytokine release but did not affect other inflammasomes. Restored expression of NLRP11, but not NLRP11 lacking the PYRIN domain (PYD), restored inflammasome activation. NLRP11 was also necessary for inflammasome responses driven by NLRP3 mutations that cause cryopyrin-associated periodic syndrome (CAPS). Because NLRP11 is not expressed in mice, our observations emphasize the specific complexity of inflammasome regulation in humans.

Hydroxycarboxylic acid receptor 3 and GPR84 - Two metabolite-sensing G protein-coupled receptors with opposing functions in innate immune cells.

G protein-coupled receptors (GPCRs) are key regulatory proteins of immune cell function inducing signaling in response to extracellular (pathogenic) stimuli. Although unrelated, hydroxycarboxylic acid receptor 3 (HCA3) and GPR84 share signaling via Gαi/o proteins and the agonist 3-hydroxydecanoic acid (3HDec). Both receptors are abundantly expressed in monocytes, macrophages and neutrophils but have opposing functions in these innate immune cells. Detailed insights into the molecular mechanisms and signaling components involved in immune cell regulation by GPR84 and HCA3 are still lacking. Here, we report that GPR84-mediated pro-inflammatory signaling depends on coupling to the hematopoietic cell-specific Gα15 protein in human macrophages, while HCA3 exclusively couples to Gαi protein. We show that activated GPR84 induces Gα15-dependent ERK activation, increases intracellular Ca2+ and IP3 levels as well as ROS production. In contrast, HCA3 activation shifts macrophage metabolism to a less glycolytic phenotype, which is associated with anti-inflammatory responses. This is supported by an increased release of anti-inflammatory IL-10 and a decreased secretion of pro-inflammatory IL-1ß. In primary human neutrophils, stimulation with HCA3 agonists counteracts the GPR84-induced neutrophil activation. Our analyses reveal that 3HDec acts solely through GPR84 but not HCA3 activation in macrophages. In summary, this study shows that HCA3 mediates hyporesponsiveness in response to metabolites derived from dietary lactic acid bacteria and uncovers that GPR84, which is already targeted in clinical trials, promotes pro-inflammatory signaling via Gα15 protein in macrophages.

Roux-en-Y gastric bypass contributes to weight loss-independent improvement in hypothalamic inflammation and leptin sensitivity through gut-microglia-neuron-crosstalk.

OBJECTIVE: Hypothalamic inflammation and endoplasmic reticulum (ER) stress are extensively linked to leptin resistance and overnutrition-related diseases. Surgical intervention remains the most efficient long-term weight-loss strategy for morbid obesity, but mechanisms underlying sustained feeding suppression remain largely elusive. This study investigated whether Roux-en-Y gastric bypass (RYGB) interacts with obesity-associated hypothalamic inflammation to restore central leptin signaling as a mechanistic account for post-operative appetite suppression. METHODS: RYGB or sham surgery was performed in high-fat diet-induced obese Wistar rats. Sham-operated rats were fed ad libitum or by weight matching to RYGB via calorie restriction (CR) before hypothalamic leptin signaling, microglia reactivity, and the inflammatory pathways were examined to be under the control of gut microbiota-derived circulating signaling. RESULTS: RYGB, other than CR-induced adiposity reduction, ameliorates hypothalamic gliosis, inflammatory signaling, and ER stress, which are linked to enhanced hypothalamic leptin signaling and responsiveness. Mechanistically, we demonstrate that RYGB interferes with hypothalamic ER stress and toll-like receptor 4 (TLR4) signaling to restore the anorexigenic action of leptin, which most likely results from modulation of a circulating factor derived from the altered gut microbial environment upon RYGB surgery. CONCLUSIONS: Our data demonstrate that RYGB interferes with hypothalamic TLR4 signaling to restore the anorexigenic action of leptin, which most likely results from modulation of a circulating factor derived from the post-surgical altered gut microbial environment.

In cis TP53 and RAD51C pathogenic variants may predispose to sebaceous gland carcinomas.

Pathogenic variants in TP53 have been classically thought to cause Li-Fraumeni syndrome (LFS), a cancer predisposition with high risks for various childhood- and adult-onset malignancies. However, increased genetic testing has lately revealed, that pathogenic variant carriers exhibit a broader range of phenotypes and that penetrance may be dependent both on variant type and modifiers. Using next generation sequencing and short tandem repeat analysis, we identified germline pathogenic variants in TP53 and RAD51C located in cis on chromosome 17 in a 43-year-old male, who has developed a rare sebaceous gland carcinoma (SGC) but so far no tumors of the LFS spectrum. This course mirrors a Trp53-Rad51c-double-mutant cis mouse-model, which similarly develops SGC, while the characteristic Trp53-associated tumor spectrum occurs with significantly lower frequency. Therefore, we propose that co-occurent pathogenic variants in RAD51C and TP53 may predispose to SGC, reminiscent of Muir-Torre syndrome. Further, this report supports the diversity of clinical presentations associated with germline TP53 alterations, and thus, the proposed expansion of LFS to heritable TP53-related cancer syndrome.

Calcium-sensing receptor-mediated NLRP3 inflammasome response to calciprotein particles drives inflammation in rheumatoid arthritis.

Increased extracellular Ca2+ concentrations ([Ca2+]ex) trigger activation of the NLRP3 inflammasome in monocytes through calcium-sensing receptor (CaSR). To prevent extraosseous calcification in vivo, the serum protein fetuin-A stabilizes calcium and phosphate into 70-100 nm-sized colloidal calciprotein particles (CPPs). Here we show that monocytes engulf CPPs via macropinocytosis, and this process is strictly dependent on CaSR signaling triggered by increases in [Ca2+]ex. Enhanced macropinocytosis of CPPs results in increased lysosomal activity, NLRP3 inflammasome activation, and IL-1ß release. Monocytes in the context of rheumatoid arthritis (RA) exhibit increased CPP uptake and IL-1ß release in response to CaSR signaling. CaSR expression in these monocytes and local [Ca2+] in afflicted joints are increased, probably contributing to this enhanced response. We propose that CaSR-mediated NLRP3 inflammasome activation contributes to inflammatory arthritis and systemic inflammation not only in RA, but possibly also in other inflammatory conditions. Inhibition of CaSR-mediated CPP uptake might be a therapeutic approach to treating RA.

Correction: Metabolites of lactic acid bacteria present in fermented foods are highly potent agonists of human hydroxycarboxylic acid receptor 3.

[This corrects the article DOI: 10.1371/journal.pgen.1008145.].

Metabolites of lactic acid bacteria present in fermented foods are highly potent agonists of human hydroxycarboxylic acid receptor 3.

The interplay of microbiota and the human host is physiologically crucial in health and diseases. The beneficial effects of lactic acid bacteria (LAB), permanently colonizing the human intestine or transiently obtained from food, have been extensively reported. However, the molecular understanding of how LAB modulate human physiology is still limited. G protein-coupled receptors for hydroxycarboxylic acids (HCAR) are regulators of immune functions and energy homeostasis under changing metabolic and dietary conditions. Most mammals have two HCAR (HCA1, HCA2) but humans and other hominids contain a third member (HCA3) in their genomes. A plausible hypothesis why HCA3 function was advantageous in hominid evolution was lacking. Here, we used a combination of evolutionary, analytical and functional methods to unravel the role of HCA3 in vitro and in vivo. The functional studies included different pharmacological assays, analyses of human monocytes and pharmacokinetic measurements in human. We report the discovery of the interaction of D-phenyllactic acid (D-PLA) and the human host through highly potent activation of HCA3. D-PLA is an anti-bacterial metabolite found in high concentrations in LAB-fermented food such as Sauerkraut. We demonstrate that D-PLA from such alimentary sources is well absorbed from the human gut leading to high plasma and urine levels and triggers pertussis toxin-sensitive migration of primary human monocytes in an HCA3-dependent manner. We provide evolutionary, analytical and functional evidence supporting the hypothesis that HCA3 was consolidated in hominids as a new signaling system for LAB-derived metabolites.

Antibacterial Silicon Oxide Thin Films Doped with Zinc and Copper Grown by Atmospheric Pressure Plasma Chemical Vapor Deposition.

Zn-doped and Cu-doped SiOx films were synthesized by atmospheric pressure plasma chemical vapor deposition to study their antibacterial efficiency against Gram-negative Escherichia coli and their cytotoxic effect on the growth of mouse cells. Zn-rich and Cu-rich particles with diameters up to several microns were found to be homogeneously distributed within the SiOx films. For both doping elements, bacteria are killed within the first three hours after exposure to the film surface. In contrast, mouse cells grow well on the surfaces of both film types, with a slight inhibition present only after the first day of exposure. The obtained results indicate that the films show a high potential for use as effective antibacterial surfaces for medical applications.

Dendritic Cells Regulate GPR34 through Mitogenic Signals and Undergo Apoptosis in Its Absence.

Dendritic cells (DCs) are specifically equipped with the G protein-coupled receptor 34 (GPR34). Tight regulation of GPR34 gene expression seems highly important for proper immunological functions, because the absence of this receptor leads to an alteration of the immune response, whereas overexpression was reported to be involved in neuroinflammation. However, the regulatory mechanism of GPR34 expression has not yet been investigated. Whole-transcriptome RNA sequencing analysis from spleens and DCs of GPR34 knockout and wild-type mice, combined with protein-protein interaction data, revealed functional modules affected by the absence of this receptor. Among these, NF-κB, MAPK, and apoptosis-signaling pathways showed high significance. Using murine DCs we experimentally show that NF-κB and MAPK pathways are involved in the downregulation of GPR34. DCs lacking GPR34 have a higher caspase-3/7 activity and increased apoptosis levels. Our study reveals a novel role of GPR34 in the fate of DCs and identifies a regulatory mechanism that could be relevant for treatment of GPR34-overexpressing pathologies, such as neuroinflammatory or cancer conditions.

Experiences of Domestic and School Violence Among Child and Adolescent Psychiatric Outpatients.

The experience of cumulative childhood adversities, such as exposure to domestic violence or abuse by caregivers, has been described as risk factor for poor mental health outcomes in adolescence and adulthood. We performed an investigation of experience of violence in all patients aged 6 to 20 years who had consulted the Department of Child and Adolescent Psychiatry, Medical University of Vienna, as outpatients during the period of one year. We were using the Childhood Trauma Interview (CTI) in order to obtain information on the kind of violence. Seventy-five percent of all patients had reported experiences of violence. These youth were significantly more often involved in acts of school violence, thus a significant correlation between experience of domestic violence and violence at school could be revealed. The results of our study emphasize the need for interventions preventing violence both in domestic and in school environments.

Challenges of river basin management: Current status of, and prospects for, the River Danube from a river engineering perspective.

In the Danube River Basin multiple pressures affect the river system as a consequence of river engineering works, altering both the river hydrodynamics and morphodynamics. The main objective of this paper is to identify the effects of hydropower development, flood protection and engineering works for navigation on the Danube and to examine specific impacts of these developments on sediment transport and river morphology. Whereas impoundments are characterised by deposition and an excess of sediment with remobilisation of fine sediments during severe floods, the remaining five free flowing sections of the Danube are experiencing river bed erosion of the order of several centimetres per year. Besides the effect of interruption of the sediment continuum, river bed degradation is caused by an increase in the sediment transport capacity following an increase in slope, a reduction of river bed width due to canalisation, prohibition of bank erosion by riprap or regressive erosion following base level lowering by flood protection measures and sediment dredging. As a consequence, the groundwater table is lowered, side-arms are disconnected, instream structures are lost and habitat quality deteriorates affecting the ecological status of valuable floodplains. The lack of sediments, together with cutting off meanders, leads also to erosion of the bed of main arms in the Danube Delta and coastal erosion. This paper details the causes and effects of river engineering measures and hydromorphological changes for the Danube. It highlights the importance of adopting a basin-wide holistic approach to river management and demonstrates that past management in the basin has been characterised by a lack of integration. To-date insufficient attention has been paid to the wide-ranging impacts of river engineering works throughout the basin: from the basin headwaters to the Danube Delta, on the Black Sea coast. This highlights the importance of new initiatives that seek to advance knowledge exchange and knowledge transfer within the basin to reach the goal of integrated basin management.

HPat a decapping activator interacting with the miRNA effector complex.

Animal miRNAs commonly mediate mRNA degradation and/or translational repression by binding to their target mRNAs. Key factors for miRNA-mediated mRNA degradation are the components of the miRNA effector complex (AGO1 and GW182) and the general mRNA degradation machinery (deadenylation and decapping enzymes). The CCR4-NOT1 complex required for the deadenylation of target mRNAs is directly recruited to the miRNA effector complex. However, it is unclear whether the following decapping step is only a consequence of deadenylation occurring independent of the miRNA effector complex or e.g. decapping activators can get recruited to the miRNA effector complex. In this study we performed split-affinity purifications in Drosophila cells and provide evidence for the interaction of the decapping activator HPat with the miRNA effector complex. Furthermore, in knockdown analysis of various mRNA degradation factors we demonstrate the importance of NOT1 for this interaction. This suggests that deadenylation and/or the recruitment of NOT1 protein precedes the association of HPat with the miRNA effector complex. Since HPat couples deadenylation and decapping, the recruitment of HPat to the miRNA effector complex provides a mechanism to commit the mRNA target for degradation.

Awareness and knowledge of child abuse amongst physicians - a descriptive study by a sample of rural Austria.

This study with a selected sample of physicians was conducted to assess their awareness and knowledge of child abuse. Two thirds (66.7%) of all participants confirmed contact with obviously abused children in the course of their professional life, whereas 87.3% did not report any prior education or training in that field. In relation to general practitioners, pediatricians had significantly more contacts with abused children (p = 0.021) and more prior education (p = 0.012). Results indicate that physicians in rural regions of Austria possess basic knowledge. Better training and further specialization is needed to facilitate diagnosing, enhance reporting, strengthen cooperation with experts and reduce fears when handling abuse victims. Austria is a rich country with excellent health care and competitive research structures. However, child abuse research in Austria still has to fill gaps in order to keep up with international developments.

The decapping activator HPat a novel factor co-purifying with GW182 from Drosophila cells.

miRNAs post-transcriptionally regulate gene expression in many eukaryotes and thereby affect a wide range of biological processes. GW182 is a key factor in translation repression and mRNA degradation by miRNAs. In this study we investigate the potential interaction of GW182 and translation or mRNA degradation factors in Drosophila S2 cells. We have identified the decapping activator HP at as a novel factor co-purifying with GW182. Furthermore, we show that the C-terminal domain of GW182, important for gene silencing, is sufficient to form a complex with HP at. Our findings implicate a potential interaction of the miRNA effector component GW182 with the decapping machinery.

[Referral success to psychotherapy of patients with personality disorders - therapeutic consequences]. / Uberweisungserfolg in Psychotherapie bei Patienten mit Persönlichkeitsstörungen - Therapeutische Konsequenzen.

BACKGROUND: Psychotherapy plays an important role in the treatment of patients suffering from a personality disorder. It is known that many patients with personality disorders do not take up psychotherapy or drop out of treatment prematurely. The aim of the present study was the detection of factors in patients with personality disorders which influence the referral to psychotherapy. METHODS: Personality characteristics (socio-demographic parameters, affect experience and regulation, quality of object relations, character traits, level of interpersonal problems) of 297 patients of a psychoanalytic-psychotherapeutic outpatient clinic were assessed. Their influence on therapy engagement were analysed by means of logistic regressions. RESULTS: Within univariate analysis certain personality traits (mature psychological functioning vs. negativistic personality features) showed predictive power. The multivariate analysis identified the patients' educational level as the principal indicator for psychotherapy utilization. CONCLUSIONS: Consequences for diagnostic initial interviews in connection with the role of the educational level for the therapeutic alliance are discussed. Further, the impact of economic aspects on therapy engagement is discussed.

[Predictive power on therapy engagement in personality disorders: SWAP- 200 versus SCID-II]. / Voraussagekraft für die Therapie-Inanspruchnahme bei Persönlichkeitsstörungen: SWAP-200 und SKID-II im Vergleich.

The study compares the predictive power of the Shedler-Westen-Assessment Procedure-200 with the Structured Clinical Interview for DSM-IV on engagement in (psychoanalytic) psychotherapy within 297 patients with personality disorders in a 4-year-follow-up. Multinomial logistic regression showed small differences between the prediction rates in the cross-validated data. Both instruments showed clinically useful prediction rates for treatment rejecters: SWAP scales led to correct predictions with dysphoric traits as semi-stable predictors for rejecters, while SCID scales led to correct predictions with Negativistic, Depressive and Schizotypal PD as stable predictors. Results are discussed under the aspect of advantages and disadvantages of the SWAP-200 diagnostic procedure, which includes the assessment of affect-experience, defence-organisation, and object-relation-style.

[Students' attitudes towards psychotherapy: changes after a course on psychic functions in health and illness]. / Einstellungen von Medizinstudenten zu Psychotherapie: Veränderungen nach Unterricht über Psychische Funktionen in Gesundheit und Krankheit.

OBJECTIVES: Little is known about the attitude of medical students towards psychotherapy and the influence of lectures on psychotherapy during the medical curriculum on these attitudes. METHODS: 2 years in a row, medical students of the Medical University of Vienna in their fourth academic year were asked regarding their attitudes, connotations and associations towards psychotherapy before and after a 5-week psychotherapeutic course. RESULTS: The attitudes of the medical students were predominantly positive, while some prejudices could be found. The students' sex and prior experience with psychotherapy influenced their attitudes. The attitudes turned out to be relatively resistant to change through teaching. CONCLUSIONS: Reasons for the small influence of the training program on the students' attitudes and possible improvements of the course (small lecture groups) are discussed.

Psychosomatic medicine in primary care: influence of training.

BACKGROUND: General practitioners (GPs) are often confronted with patients presenting somatic symptoms presumed to be decisively modulated by psychosocial factors. OBJECTIVES: We aimed to explore GPs' reported clinical routine in dealing with these patients according to the GPs' level of training in psychosomatic medicine. METHODS: A structured postal questionnaire survey was conducted among all Austrian GPs with a standardized training background in psychosomatic medicine (three levels of training; duration between one and six years) as well as in a random national sample of Austrian GPs without such training, resulting in four study subgroups. RESULTS: Respondents estimated that between 20% and 40% of their patients presenting somatic symptoms need psychosocial factors to be addressed. Study subgroups differed significantly concerning their reported diagnostic and therapeutic routine behavior patterns. Some diagnostic approaches such as clarification of lay etiology increased linearly with the level of training. The proportion of patients receiving corresponding treatment in the GP's own practice was also reported to increase with the level of training (no training: 35%, levels one and two: 46%, level three: 54%), although all subgroups estimated that over 20% of patients do not receive any corresponding treatment. CONCLUSIONS: Results point at the clinical relevance of a general training in psychosomatic medicine in primary care. They also suggest specific training effects that need to be substantiated in observational studies.