Illustrating the pathway from affect to somatic symptom: the Affective Picture Paradigm.

High levels of somatic symptom distress represent a core component of both mental and physical illness. The exact aetiology and pathogenesis of this transdiagnostic phenomenon remain largely unknown. The Affective Picture Paradigm (APP) represents an innovative experimental paradigm to study somatic symptom distress. Based on the HiTOP framework and a population-based sampling approach, associations between facets of somatic symptom distress and symptoms induced by the APP were explored in two studies (N1 = 201; N2 = 254) using structural equation bi-factor models. Results showed that the APP effect was significantly positively correlated with general somatic symptom distress (PHQ-15, HiTOP), cardio-respiratory symptoms (PHQ-15), as well as difficulties identifying feelings. In conclusion, negative affective cues in the APP can elicit somatic symptoms, particularly in people with higher levels of somatic symptom distress. Difficulties identifying feelings might contribute to this phenomenon. Results are compatible with a predictive processing account of somatic symptom perception.

Doping in Paralympic sport: perceptions, responsibility and anti-doping education experiences from the perspective of Paralympic athletes and parasport coaches.

Introduction: Limited effort has been invested in understanding doping in Paralympic sport. The limited evidence that exists suggests that factors influencing doping in parasport are similar to Olympic sport. However, based on the design and nature of the previous studies, where methods have been mostly limited to qualitative data and prevalence numbers, further research is warranted to extend previous findings. Methods: Informed by current evidence from Paralympic and Olympic sport, we aimed to investigate (1) para-athletes' perceptions of Anti-Doping Rule Violations (ADRVs) and responsibility for them, (2) descriptive norms for doping in parasport (3) perceptions of anti-doping education and legitimacy of anti-doping authorities, and (4) coach engagement in doping prevention and levels of doping confrontation efficacy using a quantitative survey approach. Results: In total, valid survey responses from 126 Paralympic athletes and 35 coaches from four countries (Germany, Austria, Switzerland, UK) were analysed for experience with anti-doping, descriptive norms, anti-doping education, perceived legitimacy, knowledge, and doping confrontation efficacy (coaches only). Across both athletes and coaches, the level of education was generally good and doping willingness was low. Classification cheating was considered a form of doping and seems to be an important issue for athletes and coaches, especially within the UK sample. For 33.3% of the athletes, doping control was their first experience with anti-doping. Coaches' engagement with doping prevention activities and their perceived efficacy to confront doping-related matters appears to be higher compared to Olympic coaches' samples. Discussion: Sport organisations/NADOs in Paralympic sport could use synergies with those organisations in Olympic sport, adopting similar approaches to anti-doping education, also focusing on a balanced communication of doping prevalence numbers and testing figures. Efforts to ensure athletes are educated about anti-doping before they are tested should be upheld. It seems that in para sport, different compared to able-bodied coaches, anti-doping organizations do not have to convince the coaches about their roles (i.e., being responsible for anti-doping education) anymore but can directly build on these resources. Overall, it seems that there are few differences between parasport and able-bodied sports and thus responsible organisations could use the existing programmes in Olympic sport and only adapt special content (e.g., boosting) which is unique to Paralympic athletes.

Confined migration promotes cancer metastasis through resistance to anoikis and increased invasiveness.

Mechanical stress is known to fuel several hallmarks of cancer, ranging from genome instability to uncontrolled proliferation or invasion. Cancer cells are constantly challenged by mechanical stresses not only in the primary tumour but also during metastasis. However, this latter has seldom been studied with regards to mechanobiology, in particular resistance to anoikis, a cell death programme triggered by loss of cell adhesion. Here, we show in vitro that migrating breast cancer cells develop resistance to anoikis following their passage through microporous membranes mimicking confined migration (CM), a mechanical constriction that cancer cells encounter during metastasis. This CM-induced resistance was mediated by Inhibitory of Apoptosis Proteins, and sensitivity to anoikis could be restored after their inhibition using second mitochondria-derived activator of caspase (SMAC) mimetics. Anoikis-resistant mechanically stressed cancer cells displayed enhanced cell motility and evasion from natural killer cell-mediated immune surveillance, as well as a marked advantage to form lung metastatic lesions in mice. Our findings reveal that CM increases the metastatic potential of breast cancer cells.

The circle of life: Phases of podosome formation, turnover and reemergence.

Podosomes are highly dynamic actin-rich structures in a variety of cell types, especially monocytic cells. They fulfill multiple functions such as adhesion, mechanosensing, or extracellular matrix degradation, thus allowing cells to detect and respond to a changing environment. These abilities are based on an intricate architecture that enables podosomes to sense mechanical properties of their substratum and to transduce them intracellularly in order to generate an appropriate cellular response. These processes are enabled through the tightly orchestrated interplay of more than 300 different components that are dynamically recruited during podosome formation and turnover. In this review, we discuss the different phases of the podosome life cycle and the current knowledge on regulatory factors that impact on the genesis, activity, dissolution and reemergence of podosomes. We also highlight mechanoregulatory processes that become important during these different stages, on the level of individual podosomes, and also at podosome sub- and superstructures.

Vacuum-Induced Tamponade for Treatment of Postpartum Hemorrhage.

BACKGROUND: Postpartum hemorrhage is a main cause of maternal mortality worldwide, with rising incidence, thus demanding new treatment approaches. Intrauterine balloon systems with application of intrauterine vacuum are a promising new method. METHOD: All women treated with vacuum-induced tamponade using a modified balloon system were included in this single-center study. Aiming to reduce uterine size for control of postpartum hemorrhage, the intrauterine balloon was filled to 50-100 mL and connected to a vacuum device. Success rate of vacuum-induced tamponade, defined as no need for additional interventional treatment, was analyzed by etiology of postpartum hemorrhage and time period of use. EXPERIENCE: Vacuum-induced tamponade was applied in 66 women. Success rate was 86% in women with uterine atony (n=44) and 73% in women with postpartum hemorrhage due to placental pathology (n=22). Success rate improved over the study period, culminating in a success rate of 100% in women with postpartum hemorrhage due to uterine atony in the second half of the observation period (n=22). CONCLUSION: This observational study supports our pathophysiologic understanding of uterine atony: to treat an atonic uterus, uterine volume must be reduced, leading to coiling of the uterine spiral arteries and, hence, reduced blood loss.

GC-MS Studies on Derivatization of Creatinine and Creatine by BSTFA and Their Measurement in Human Urine.

In consideration of its relatively constant urinary excretion rate, creatinine (2-amino-1-methyl-5H-imidazol-4-one, MW 113.1) in urine is a useful endogenous biochemical parameter to correct the urinary excretion rate of numerous endogenous and exogenous substances. Reliable measurement of creatinine by gas chromatography (GC)-based methods requires derivatization of its amine and keto groups. Creatinine exists in equilibrium with its open form creatine (methylguanidoacetic acid, MW 131.1), which has a guanidine and a carboxylic group. Trimethylsilylation and trifluoroacetylation of creatinine and creatine are the oldest reported derivatization methods for their GC-mass spectrometry (MS) analysis in human serum using flame- or electron-ionization. We performed GC-MS studies on the derivatization of creatinine (d0-creatinine), [methylo-2H3]creatinine (d3-creatinine, internal standard) and creatine (d0-creatine) with N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) using standard derivatization conditions (60 min, 60 °C), yet in the absence of any base. Reaction products were characterized both in the negative-ion chemical ionization (NICI) and in the positive-ion chemical ionization (PICI) mode. Creatinine and creatine reacted with BSTFA to form several derivatives. Their early eluting N,N,O-tris(trimethylsilyl) derivatives (8.9 min) were found to be useful for the precise and accurate measurement of the sum of creatinine and creatine in human urine (10 µL, up to 20 mM) by selected-ion monitoring (SIM) of m/z 271 (d0-creatinine/d0-creatine) and m/z 274 (d3-creatinine) in the NICI mode. In the PICI mode, SIM of m/z 256, m/z 259, m/z 272 and m/z 275 was performed. BSTFA derivatization of d0-creatine from a freshly prepared solution in distilled water resulted in formation of two lMate-eluting derivatives (14.08 min, 14.72 min), presumably creatinyl-creatinine, with the creatininyl residue existing in its enol form (14.08 min) and keto form (14.72 min). Our results suggest that BSTFA derivatization does not allow specific analysis of creatine and creatinine by GC-MS. Preliminary analyses suggest that pentafluoropropionic anhydride (PFPA) is also not useful for the measurement of creatinine in the presence of creatine. Both BSTFA and PFPA facilitate the conversion of creatine to creatinine. Specific measurement of creatinine in urine is possible by using pentafluorobenzyl bromide in aqueous acetone.

Geographical heterogeneity of doping-related knowledge, beliefs and attitude among 533 Youth Olympics participants.

OBJECTIVES: Doping-related knowledge, beliefs and attitude influence adolescent athletes' susceptibility to prohibited performance-enhancing substances. They might be modified by different cultural backgrounds. This study's aim was to analyse the geographical heterogeneity of doping-related knowledge, beliefs and attitude among adolescent elite athletes. DESIGN: Cross-sectional study. METHODS: A questionnaire was distributed to athletes participating in the Winter Youth Olympic Games 2020 in Switzerland. Main outcomes ('subjective and actual knowledge', 'beliefs' and 'attitude') were stratified for athletes' region of origin. Geographical heterogeneity was tested with a two-way analysis of variance, and two multiple regression analyses were conducted to assess independent associations of knowledge, age and athletes' geographical region with doping-related beliefs and attitude. RESULTS: 533 athletes (54% females, mean age: 16.0 ±â€¯1.0 years), completed the questionnaire (response rate: 33%). Actual knowledge was moderate-to-good (9.2 ±â€¯2.9 correct answers out of 13), and scores of attitude and beliefs showed favourable patterns. Considerable geographical heterogeneity was found for knowledge (p < 0.001), beliefs (p = 0.004) and attitude (p < 0.001). Higher subjective knowledge and actual knowledge were favourably associated with attitude (ß = -0.096, p = 0.049; ß = -0.316, p < 0.001) and beliefs (ß = 0.120, p = 0.016; ß = 0.212, p < 0.001), independent of age and geographical region. CONCLUSIONS: This study demonstrates considerable geographical heterogeneity of doping-related knowledge, beliefs and attitude, which are three essential target factors of doping prevention in adolescent elite athletes. This evidence should encourage medical doctors and other professionals to change their educative anti-doping approach from teaching knowledge about negative consequences into investigating and forming a young athlete's mind-set.

Caspase-8 deficiency induces a switch from TLR3 induced apoptosis to lysosomal cell death in neuroblastoma.

In cancer cells only, TLR3 acquires death receptor properties by efficiently triggering the extrinsic pathway of apoptosis with Caspase-8 as apical protease. Here, we demonstrate that in the absence of Caspase-8, activation of TLR3 can trigger a form of programmed cell death, which is distinct from classical apoptosis. When TLR3 was activated in the Caspase-8 negative neuroblastoma cell line SH-SY5Y, cell death was accompanied by lysosomal permeabilization. Despite caspases being activated, lysosomal permeabilization as well as cell death were not affected by blocking caspase-activity, positioning lysosomal membrane permeabilization (LMP) upstream of caspase activation. Taken together, our data suggest that LMP with its deadly consequences represents a "default" death mechanism in cancer cells, when Caspase-8 is absent and apoptosis cannot be induced.

Restoration of teeth affected by molar-incisor hypomineralisation: a systematic review.

The objective was to systematically analyse clinical studies on restorative procedures for teeth affected by molar-incisor hypomineralisation (MIH). The databases PubMed, Embase, and Cochrane Library were searched. Only retrospective and prospective clinical studies dealing with sealing or restoration of MIH-affected teeth were included. The language was restricted to English or German. Thirteen of 36 potentially eligible studies were included focusing on the following subjects: extension of enamel preparation, adhesive procedures prior to restoration, application of fissure sealants as well as restoration with conventional glass ionomer cements (GIC), resin modified glass ionomer cements (RMGIC), resin composites, and indirect restorations. Seven clinical studies were controlled trials. However, only two included MIH-unaffected teeth as control. No meta-analysis was performed due to the heterogeneity of study designs (e.g. severity of MIH or the restorative materials investigated). Based on the present analysis, the annual failure rates were in average 21% for fissure sealants, 22% for GIC, 1-6% for RMGIC, 13-32% for resin composites, and 0-7% for indirect restorations. In summary, only few tendencies can be deduced from this review at a low level of evidence (number of studies): 1) preparation margins in sound enamel seem to be superior to preparations in hypomineralised enamel (1 study), 2) RMGIC seems to be superior to GIC (3 studies), 3) resin composites may be used for restoring all severities of MIH (7 studies) with self-etch and etch-and-rinse adhesive systems generally not performing differently (3 studies), and 4) in cases of severe MIH, indirect restorations showed a good clinical success (4 studies).

Sometimes even apoptosis fails: implications for cancer.

Apoptosis is vital for the correct morphogenesis of multi-cellular organisms. However, like most physiological programs, the cell's ability to commit suicide is hijacked by cancer in its own proliferative and invasive interest. We recently showed that inefficient execution of apoptosis (or failed apoptosis) is used by cancer to boost invasiveness.

Nucleobindin-1 regulates ECM degradation by promoting intra-Golgi trafficking of MMPs.

Matrix metalloproteinases (MMPs) degrade several ECM components and are crucial modulators of cell invasion and tissue organization. Although much has been reported about their function in remodeling ECM in health and disease, their trafficking across the Golgi apparatus remains poorly understood. Here we report that the cis-Golgi protein nucleobindin-1 (NUCB1) is critical for MMP2 and MT1-MMP trafficking along the Golgi apparatus. This process is Ca2+-dependent and is required for invasive MDA-MB-231 cell migration as well as for gelatin degradation in primary human macrophages. Our findings emphasize the importance of NUCB1 as an essential component of MMP transport and its overall impact on ECM remodeling.

Status of waste-to-energy in Germany, Part I - Waste treatment facilities.

This study gives a detailed overview over the German waste-to-energy sector in 2015. The aim is to quantify the available treatment capacities and the energetic potential of waste in Germany. The work is based on an extensive data collection and evaluation, both from literature sources as well as from a survey among operators of waste treatment plants. The present Part I, gives an overview of all treatment facilities in Germany that convert waste into energy. It was found that in total, almost 320 PJ of end energy are produced in German waste treatment plants: 225 PJ a-1 of heat; and 90 PJ a-1 of electricity. This is a share of about 3.7% of the German end energy consumption.

Cisplatin unleashes Toll-like receptor 3-mediated apoptosis through the downregulation of c-FLIP in malignant mesothelioma.

Toll-like receptor 3 (TLR3) is an immune receptor that behaves like a death receptor in tumor cells, thereby providing an original target for cancer therapy. The therapeutic potential of TLR3 targeting in malignant mesothelioma, an aggressive and incurable neoplasia of the pleura and peritoneum, has so far not been addressed. We investigated TLR3 expression and sensitivity of human mesothelioma cell lines to the synthetic dsRNA Poly(I:C), alone or in combination with cisplatin, the gold standard chemotherapy in mesothelioma. Activation of TLR3 by Poly(I:C) induced apoptosis of 4/8 TLR3-positive cell lines but not of TLR3-negative cell lines. The combined cisplatin/Poly(I:C) treatment enhanced apoptosis of 3/4 Poly(I:C)-sensitive cell lines and overcame resistance to Poly(I:C) or cisplatin alone in 2/4 cell lines. Efficacy of the combined treatment relied on cisplatin-induced downregulation of c-FLIP, the main regulator of the extrinsic apoptotic pathway, leading to an enhanced caspase-8-mediated pathway. Of note, 6/6 primary cell samples isolated from patients with peritoneal mesothelioma expressed TLR3. Patient-derived cells were sensitive to Poly(I:C) alone while the combined cisplatin/Poly(I:C) treatment induced dramatic cell death. Our findings demonstrate that TLR3 targeting in combination with cisplatin presents an innovative therapeutic strategy in mesothelioma.

Toll-like receptor 3 downregulation is an escape mechanism from apoptosis during hepatocarcinogenesis.

BACKGROUND & AIMS: Low levels of toll-like receptor 3 (TLR3) in patients with hepatocellular carcinoma (HCC) are associated with poor prognosis, primarily owing to the loss of inflammatory signaling and subsequent lack of immune cell recruitment to the liver. Herein, we explore the role of TLR3-triggered apoptosis in HCC cells. METHODS: Quantitative reverse transcription PCR, western blotting, immunohistochemistry and comparative genomic hybridization were used to analyze human and mouse HCC cell lines, as well as surgically resected primary human HCCs, and to study the impact of TLR3 expression on patient outcomes. Functional analyses were performed in HCC cells, following the restoration of TLR3 by lentiviral transduction. The role of TLR3-triggered apoptosis in HCC was analyzed in vivo in a transgenic mouse model of HCC. RESULTS: Lower expression of TLR3 in tumor compared to non-tumor matched tissue was observed at both mRNA and protein levels in primary HCC, and was predictive of shorter recurrence-free survival after surgical resection in both univariate (hazard ratio [HR] 1.79; 95% CI 1.04-3.06; p = 0.03) and multivariate analyses (HR 1.73; CI 1.01-2.97; p = 0.04). Immunohistochemistry confirmed frequent downregulation of TLR3 in human and mouse primary HCC cells. None of the 6 human HCC cell lines analyzed expressed TLR3, and ectopic expression of TLR3 following lentiviral transduction not only restored the inflammatory response but also sensitized cells to TLR3-triggered apoptosis. Lastly, in the transgenic mouse model of HCC, absence of TLR3 expression was accompanied by a lower rate of preneoplastic hepatocyte apoptosis and accelerated hepatocarcinogenesis without altering the tumor immune infiltrate. CONCLUSION: Downregulation of TLR3 protects transforming hepatocytes from direct TLR3-triggered apoptosis, thereby contributing to hepatocarcinogenesis and poor patient outcome. LAY SUMMARY: Hepatocellular carcinoma (HCC) is a heterogeneous disease associated with a poor prognosis. In patients with HCC, TLR3 downregulation is associated with reduced survival. Herein, we show that the absence of TLR3 is associated with a lower rate of apoptosis, and subsequently more rapid hepatocarcinogenesis, without any change to the immune infiltrate in the liver. Therefore, the poor prognosis associated with low TLR3 expression in HCC is likely linked to tumors ability to escape apoptosis. TLR3 may become a promising therapeutic target in TLR3-positive HCC.

Stem Cell Therapy for Tendon Regeneration: Current Status and Future Directions.

In adults the healing tendon generates fibrovascular scar tissue and recovers never histologically, mechanically, and functionally which leads to chronic and to degenerative diseases. In this review, the processes and mechanisms of tendon development and fetal regeneration in comparison to adult defect repair and degeneration are discussed in relation to regenerative therapeutic options. We focused on the application of stem cells, growth factors, transcription factors, and gene therapy in tendon injury therapies in order to intervene the scarring process and to induce functional regeneration of the lesioned tissue. Outlines for future therapeutic approaches for tendon injuries will be provided.

Neuron-specific inactivation of Wt1 alters locomotion in mice and changes interneuron composition in the spinal cord.

Locomotion is coordinated by neuronal circuits of the spinal cord. Recently, dI6 neurons were shown to participate in the control of locomotion. A subpopulation of dI6 neurons expresses the Wilms tumor suppressor gene Wt1. However, the function of Wt1 in these cells is not understood. Here, we aimed to identify behavioral changes and cellular alterations in the spinal cord associated with Wt1 deletion. Locomotion analyses of mice with neuron-specific Wt1 deletion revealed a slower walk with a decreased stride frequency and an increased stride length. These mice showed changes in their fore-/hindlimb coordination, which were accompanied by a loss of contralateral projections in the spinal cord. Neonates with Wt1 deletion displayed an increase in uncoordinated hindlimb movements and their motor neuron output was arrhythmic with a decreased frequency. The population size of dI6, V0, and V2a neurons in the developing spinal cord of conditional Wt1 mutants was significantly altered. These results show that the development of particular dI6 neurons depends on Wt1 expression and that loss of Wt1 is associated with alterations in locomotion.

Nuclear RIPK3 and MLKL contribute to cytosolic necrosome formation and necroptosis.

Necroptotic signaling converges in the assembly of a cytosolic signaling platform, the necrosome, with the activation of its downstream effector, MLKL. RIPK1 and RIPK3, key components of the necrosome, act as signaling intermediates for the activation of MLKL. We report that RIPK3 and MLKL continuously shuttle between the nucleus and the cytoplasm, whereas RIPK1 is constitutively present in both compartments. During TNF-induced necroptosis, nuclear RIPK1 becomes ubiquitinated, after which nuclear MLKL becomes phosphorylated and oligomerized. Pharmacological inhibition of the nuclear export machinery leads to retention of RIPK3 and MLKL in the nucleus, prevents the nucleation of cytosolic RIPK3/MLKL oligomerization, and reduces cell death. Our results suggest that passage of necroptotic signaling components through the nucleus is a mechanism for regulating cytosolic necrosome formation and consequently necroptotic cell death.

Performance in Sound-Symbol Learning Predicts Reading Performance 3 Years Later.

To master the task of reading, children need to acquire a coding system representing speech as a sequence of visual symbols. Recent research suggested that performance in the processing of artificial script that relies on the association of sound and symbol may be associated with reading skill. The current longitudinal study examined the predictive value of a preschool sound-symbol paradigm (SSP) of reading performance 3 years later. The Morse-like SSP, IQ, and letter knowledge (LK) was assessed in young preschool children. Reading outcome measures were examined 3 years later. Word reading, pseudoword reading, and reading comprehension were predicted with age, IQ, LK, and SSP. The results showed that SSP substantially predicted reading fluency and reading comprehension 3 years later. For reading fluency measures, the influence of further predictor variables was not significant and SSP served as a sole predictor. Reading comprehension was best explained by SSP and age. The amount of variance SSP explained in reading 3 years later was remarkably high, with an explained variance between 63 and 82%, depending on the outcome reading variable. SSP turned out to be a substantial predictor of later reading performance in a language with statistically reliable spelling-to-sound relations. As LK is highly dependent on educational support, we assume that children in our socioeconomically diverse sample did not have much opportunity to acquire LK in their home environment. In contrast, the SSP challenges students to acquire new spelling-to-sound relations, simulating a core aspect of natural reading acquisition. Future work will test this paradigm in less transparent languages like English and explore its potential as a future standard assessment in the study of early reading development.

Release of c-FLIP brake selectively sensitizes human cancer cells to TLR3-mediated apoptosis.

Toll-like receptor 3 (TLR3) mediates innate immune responses by sensing viral dsRNA, but also induces apoptosis selectively in cancer cells. Our analysis by immunohistochemistry revealed that TLR3 is frequently overexpressed in 130 non-small cell lung cancer (NSCLC) patients' samples compared with normal bronchial epithelium (P < 0.0001, Mann-Whitney test), supporting the therapeutic potential of TLR3 ligand for this type of cancer. However, a proportion of TLR3-expressing cancer cell lines, including NSCLC, remain resistant to TLR3-mediated apoptosis, and the underlying mechanism of resistance remains unclear. We here investigated the molecular basis conferring resistance to non-transformed vs. transformed cells against TLR3-mediated cell death. In non-transformed epithelial cells cellular FLICE-like inhibitory protein (c-FLIP) and cellular Inhibitor of APoptosis (cIAPs) ubiquitin ligases exerted an efficient double brake on apoptosis signaling. In contrast, releasing only one of these two brakes was sufficient to overcome the resistance of 8/8 cancer cell lines tested. Remarkably, the release of the c-FLIP, but not cIAPs, brake only results in the sensitization of all human cancer cells to TLR3-mediated apoptosis. Taking advantage of the difference between transformed and non-transformed cells, we developed a rational strategy by combining the chemotherapeutic agent paclitaxel, which decreases c-FLIP expression, with TLR3 ligand. This combination was highly synergistic for triggering apoptosis in cancer cells but not in non-transformed cells. In vivo, the combination of paclitaxel with dsRNA delayed tumor growth and prolonged survival in a mouse xenograft lung tumor model. In conclusion, combining the release of the c-FLIP brake with TLR3 ligand synergizes to selectively kill cancer cells, and could represent an efficient and safe therapy against TLR3-expressing cancers such as NSCLC.