Paradigm of biased PAR1 (protease-activated receptor-1) activation and inhibition in endothelial cells dissected by phosphoproteomics

Thrombin is the key serine protease of the coagulation cascade and mediates cellular responses by activation of PARs (protease-activated receptors). The predominant thrombin receptor is PAR1, and in endothelial cells (ECs), thrombin dynamically regulates a plethora of phosphorylation events. However, it has remained unclear whether thrombin signaling is exclusively mediated through PAR1. Furthermore, mechanistic insight into activation and inhibition of PAR1-mediated EC signaling is lacking. In addition, signaling networks of biased PAR1 activation after differential cleavage of the PAR1 N terminus have remained an unresolved issue. Here, we used a quantitative phosphoproteomics approach to show that classical and peptide activation of PAR1 induce highly similar signaling, that low thrombin concentrations initiate only limited phosphoregulation, and that the PAR1 inhibitors vorapaxar and parmodulin-2 demonstrate distinct antagonistic properties. Subsequent analysis of the thrombin-regulated phosphosites in the presence of PAR1 inhibitors revealed that biased activation of PAR1 is not solely linked to a specific G-protein downstream of PAR1. In addition, we showed that only the canonical thrombin PAR1 tethered ligand induces extensive early phosphoregulation in ECs. Our study provides detailed insight in the signaling mechanisms downstream of PAR1. Our data demonstrate that thrombin-induced EC phosphoregulation is mediated exclusively through PAR1, that thrombin and thrombin-tethered ligand peptide induce similar phosphoregulation, and that only canonical PAR1 cleavage by thrombin generates a tethered ligand that potently induces early signaling. Furthermore, platelet PAR1 inhibitors directly affect EC signaling, indicating that it will be a challenge to design a PAR1 antagonist that will target only those pathways responsible for tissue pathology

Reconstruction of an Extensive Midfacial Defect Using Additive Manufacturing Techniques

Published ArticleMalignant peripheral nerve sheath tumors are extremely rare tumors arising in peripheral nerves. Only 17 cases involving the trigeminal nerve have ever been reported. These tumors have a very poor prognosis and very high rates of recurrence and metastases. Their recommended treatment involves complete tumor resection followed by radiation. This can be problematic in the head and neck region. We present a clinical case involving a 33-year-old female patient presenting with a slow-growing, exophytic mass of the anterior maxilla. Incisional biopsy and subsequent histological examination revealed a diagnosis of a malignant peripheral nerve sheath tumor. Surgical resection involved a complete maxillectomy, rhinectomy, and resection of the upper lip and aspects of the left and right cheeks. Reconstruction of the subsequent defect incorporated the placement of four zygomatic oncology implants to aid in retention of a facial prosthesis. These implants, however, were subsequently lost; and an anatomical model of the hard tissues was manufactured via 3D printing. This model was used to design and manufacture a titanium frame (customized implant) for the patient. The frame was then fixated and secured intraoperative with 21 cortical screws. A maxillary denture and silicone facial prosthesis were also made to fit onto this frame. This is the first known case where additive manufacturing, via the use of rapid prototyping and 3D printing, was employed to manufacture a facial prosthesis

Associations between the urban environment and psychotic experiences in adolescents

Objective: In 2050 two-thirds of the world's population is predicted to live in cities, which asks for a better understanding of how the urban environment affects mental health. Urbanicity has repeatedly been found to be a risk factor, in particular for psychosis. Here, we explored what factors of the urban exposome underlie the association between urban characteristics and psychotic experiences (PE) in adolescents. Methods: Participants were 815 adolescents (mean age 14.84 years, SD 0.78) from an at-risk cohort (greater Rotterdam area, the Netherlands) oversampled on their self-reported emotional and behavioral problems. We used linear regression analysis to examine the association with detailed geodata on urbanicity (surrounding address density), green space density (high and low vegetation), and mixed noise levels (road, rail, air, industry, and wind power) with PE in adolescents. Analyses were adjusted for multiple socio-economic and parental confounders. Furthermore, we explored sex-interaction effects. Results:Higher surrounding address density and low greenspace density were each independently associated with more PE (B = 0.18, 95 % CI 0.02; 0.34 and B = 0.17, 95 % CI 0.01; 0.32, respectively). High mixed noise levels were only associated with more PE in boys (B = 0.23, 95 % CI 0.01; 0.46). A sex-interaction effect was found for high urbanicity (B = −0.46, 95 % CI −0.77; −0.14) and low greenspace density (B = −0.49, 95 % CI −0.73; −0.11), illustrating that these associations with PE were specific for boys. Conclusion: Multiple characteristics of living in an urban area are associated with more PE in adolescent boys. Our observations provide leads for prevention of mental health problems via urban designing.</p

How are you coping?:Stress, coping, burnout, and aggression in forensic mental healthcare workers

IntroductionPerceived stress at work has been linked to several adverse outcomes in workers, including increased risk of burnout and aggression (e.g., anger and irritability). However, much remains unknown about factors that might mitigate the negative influences of perceived stress on workers' well-being. This study focusses on coping as a possible protective factor against perceived stress and its consequences in forensic mental healthcare workers. We aimed to identify which higher-order coping factors were present in this worker sample and to investigate whether these coping factors modify the associations between perceived stress and burnout or aggression.MethodsFor this observational survey study, 116 forensic mental healthcare workers completed questionnaires assessing changes in work situation since the start of COVID-19, perceived stress, coping, burnout symptoms, and aggression.ResultsResults from principal component analysis indicated that four higher-order coping factors could be distinguished: social support and emotional coping, positive cognitive restructuring, problem-focused coping, and passive coping. Higher perceived stress levels were associated with higher levels of both burnout and aggression in workers. Problem-focused coping was associated with less burnout symptoms in workers. Furthermore, positive cognitive restructuring was associated with less aggression in workers.DiscussionIn conclusion, problem-focused coping and positive cognitive restructuring may protect workers against burnout symptoms and aggression and these results may inform future studies on preventive interventions aimed at promoting worker's well-being

Kinase Activity Profiling of Gram-Negative Pneumonia

Pneumonia is a severe disease with high morbidity and mortality. A major causative pathogen is the Gram-negative bacterium Klebsiella (K.) pneumoniae. Kinases play an integral role in the transduction of intracellular signaling cascades and regulate a diverse array of biological processes essential to immune cells. The current study explored signal transduction events during murine Gram-negative pneumonia using a systems biology approach. Kinase activity arrays enable the analysis of 1,024 consensus sequences of protein kinase substrates. Using a kinase activity array on whole lung lysates, cellular kinase activities were determined in a mouse model of K. pneumoniae pneumonia. Notable kinase activities also were validated with phospho-specific Western blots. On the basis of the profiling data, mitogen-activated protein kinase (MAPK) signaling via p42 mitogen-activated protein kinase (p42) and p38 mitogen-activated protein kinase (p38) and transforming growth factor β (TGFβ) activity were reduced during infection, whereas v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (avian) (SRC) activity generally was enhanced. AKT signaling was represented in both metabolic and inflammatory (mitogen-activated protein kinase kinase 2 [MKK], apoptosis signal-regulating kinase/mitogen-activated protein kinase kinase kinase 5 [ASK] and v-raf murine sarcoma viral oncogene homolog B1 [b-RAF]) context. This study reaffirms the importance of classic inflammation pathways, such as MAPK and TGFβ signaling and reveals less known involvement of glycogen synthase kinase 3β (GSK-3β), AKT and SRC signaling cassettes in pneumonia

Vitamin K status and physical decline in older adults—The Longitudinal Aging Study Amsterdam

Objective: We examined the association between vitamin K status and physical functioning over 13 years in the Longitudinal Aging Study Amsterdam. Study design: Longitudinal cohort study of 633 community-dwelling adults from the Longitudinal Aging Study Amsterdam (LASA) aged 55–65 years (54% women). Main outcome measures: At baseline (2002–2003), plasma desphospho-uncarboxylated matrix Gla protein (dp-ucMGP) was measured with a sandwich ELISA as a marker of vitamin K status. The outcome measures handgrip strength, calf circumference, self-reported functional limitations and functional performance were obtained at baseline and four follow-up examinations. We used generalized estimating equations to determine the relationship between dp-ucMGP tertiles and the various outcome measurements after adjusting for potential confounders. The lowest dp-ucMGP tertile reflects a high vitamin K status and was the reference. Results: Mean dp-ucMGP was 376 ± 233 pmol/L and mean follow-up was 11.1 years. Participants showed a decline in the outcome measures over time. Compared with the lowest tertile, the highest dp-ucMGP tertile had: lower handgrip strength, 1.1 kg (95% confidence interval (−2.1, −0.1; P-trend <0.001); smaller calf circumference, −0.5 cm (−0.9 −0.1; P-trend = 0.018); and, only among women, a 0.7-point poorer functional performance score (−1.1, −0.3; P-interaction = 0.002). Dp-ucMGP was not related to self-reported functional limitations. No interaction effects between time and dp-ucMGP were observed. Conclusions: Low vitamin K status was associated with lower handgrip strength, smaller calf circumference, and, in women only, with poorer functional performance score. A low vitamin K status was however not related to the 13-year decline in these measures

Trends in Frailty and Its Association With Mortality:Results From the Longitudinal Aging Study Amsterdam, 1995-2016

The aim of this study was to investigate trends in frailty and its relationship with mortality among older adults aged 64-84 years across a period of 21 years. We used data from 1995 to 2016 from the Longitudinal Aging Study Amsterdam. A total of 7,742 observations of 2,874 respondents in the same age range (64-84 years) across 6 measurement waves were included. Frailty was measured with a 32-item frailty index, with a cutpoint of ≥0.25 to indicate frailty. The outcome measure was 4-year mortality. Generalized estimating equation analyses showed that among older adults aged 64-84 years the 4-year mortality rate declined between 1995 and 2016, while the prevalence of frailty increased. Across all measurement waves, frailty was associated with 4-year mortality (odds ratio = 2.79, 95% confidence interval: 2.39, 3.26). There was no statistically significant interaction effect between frailty and time on 4-year mortality, indicating a stable association between frailty and mortality. In more recent generations of older adults, frailty prevalence rates were higher, while excess mortality rates of frailty remained the same. This is important information for health policy-makers and clinical practitioners, showing that continued efforts are needed to reduce frailty and its negative health consequences

Mother-to-Infant Bonding in Women with Postpartum Psychosis and Severe Postpartum Depression: A Clinical Cohort Study

Mother-to-infant bonding is important for long-term child development. The aim of this study was to investigate bonding in women admitted to a Mother and Baby Unit with postpartum depression (PD, n = 64) and postpartum psychosis (PP, n = 91). Participants completed the Postpartum Bonding Questionnaire (PBQ), the Edinburgh Postnatal Depression Scale (EPDS) and the Young Mania Rating Scale (YMRS) weekly during admission. At admission, 57.1% of women with PD had impaired bonding, compared to only 17.6% of women with PP (p-value < 0.001). At discharge, only 18.2% of women with PD and 5.9% of women with PP still experienced impaired bonding (p-value = 0.02). There was a strong association between decrease of depressive and manic symptoms and improved bonding over an eight-week admission period. In a small group of women (5.7%) impaired bonding persisted despite being in remission of their psychiatric disorder. The results from our study show that impaired bonding is a more present and evidently severe problem in postpartum depression but not so much in postpartum psychosis. Treatment of depressive symptoms will improve bonding in almost all women, but clinicians should assess if impaired bonding is still present after remission because for a small group special care and treatment focused on bonding might be required

Effects of Key2Teach on students' externalising and social-emotional problem behaviours, mediated by the teacher-student relationship

Teaching students with externalising problem behaviours is difficult for teachers, as it challenges the relationship that teachers engage in with their students. In this study, effects of Key2Teach on externalising and social-emotional problem behaviours and the mediating role of conflict in the teacher-student relationship were studied using a randomised controlled trial (RCT)-design. In two cohorts, 103 teacher-student dyads and peer-students (n = 1643) were assessed two times during a school year. Fifty-three dyads received the intervention (experimental group), whereas 50 dyads received no intervention (control group). Data were collected on teacher-reported externalising and social-emotional problem behaviours in students, and on teacher-student conflict. For dyad-students, results showed a direct effect of Key2Teach on conduct problems and an indirect effect, via teacher-student conflict, for hyperactivity problems. No effects on social-emotional problems were established. For peer-students, results showed indirect effects on externalising problems and direct effects on social-emotional problems. Implications for practice and future research are discussed