Successful Development of a Flow Cytometric Assay to Analyze Cytokine-Induced Phosphorylation Pathways [CIPP] Within Peripheral Blood Leukocytes

Current strategies designed to assess cells in the peripheral blood are limited to evaluation of phenotype or delayed measurement [>6 hours] of function, usually quantifying cytokine production, cytolytic activity, or response to antigens. Furthermore, these assays often do not assess the individual cell types comprising peripheral blood, instead providing a bulk-readout for the total population. We reasoned that functional abnormalities in distinct peripheral blood cells, which act as immune sentinels capable of rapidly reacting to various injurious agents while circulating throughout the body, could serve as migratory biomarkers, reflecting pathological environs that cells experience in the setting of inflammatory states, including cancer. Two major pathways regulating immune responses are the JAK/STAT and MAPK pathways. These pathways are initiated by ligand-receptor binding, and are rapidly propagated by subsequent protein phosphorylation cascades. Additionally, these pathways are often abnormally activated within cancer cells themselves. We applied flow cytometric strategies, evaluating the brief application of cytokines in vitro to interrogate the early phosphorylation events of these signaling pathways in normal peripheral blood mononuclear cells (PBMC). Individual cytokine doses and time intervals of treatment were assessed to identify conditions useful in a clinical laboratory and as an initial goal, to induce maximal phosphorylation. Surprisingly, all of the STAT proteins assessed and ERK1/2 are maximally phosphorylated within 15 minutes in human PBMC, simply following addition of cytokines alone. Within two hours, cells typically return to their basal phosphorylation states. Confirmation of these results was achieved by Western blotting. Increased phosphorylation usually correlated with increased concentrations of individual cytokines. Furthermore, we developed conditions to enable simultaneous staining of cell surface proteins, identifiable with distinct PBMC subpopulations: CD4, CD8, CD14, CD19, and CD56, together with intracellular phosphorylated proteins. Of the permeabilizing conditions tested, 75% methanol enabled superior simultaneous detection of both cell surface and intracellular epitopes. Using this technique, we were able to identify differential responsiveness between individual cell types. These strategies will enable robust development of simple blood analyses to identify normal activation as well as impairments in STAT and MAPK signaling pathways associated with various human disease states including acute and chronic inflammatory conditions throughout clinical immunology

Foreign investors and domestic company growth: Evidence from US venture capital investments in Sweden

Do foreign venture capitalists help the domestic economy, or hamper it by slowing down growth, potentially moving economic activity away? This paper addresses this long-standing policy question by examining the differential effects of US venture capital investments on the growth of Swedish start-up companies. It finds that US venture capital results in more employment, not less. These findings continue to hold after controlling for endogenous selection effects. US investments are also accompanied by increases in local employment and start-up rates. The paper also examines effect on wages, sales, earnings, foreign subsidiaries, subsequent funding rounds, and exits. Overall there is no evidence that US venture capital investments hamper the domestic growth of Swedish companies

Aspects of outpatient palliative care in Germany - a survey among office-based physicians

Background: Currently, palliative care is widely discussed. The aim of this study is to explore different aspects of the ambulatory treatment environment of palliative care patients and its structure in Germany from the viewpoint of caring physicians. Methods: A structured questionnaire was developed and sent to 1,003 office-based physicians in Germany. Topics were the delivery and effort of care, cooperation among health care providers, the subjective assessment of existing health care structures, the reimbursement system, as well as an assessment of future developments. Sub-group analyses were conducted. Results: Three hundred fourteen questionnaires were included in the statistical analyses. Fifty percent of physicians estimate the additional time and effort for palliative care patients compared to non-palliative care patients to be 50% and more. The best cooperation is seen with caregivers; however, cooperation with psycho-social care providers should be improved. Fifty percent describe the outpatient health care situation as (very) insufficient, but improvements are expected in the future. Ninety percent assess German reimbursement structures as (very) insufficient. Conclusions: This study gives an insight into the German outpatient health care setting for palliative care patients. Palliative care patients have higher and more complex needs with respect to health care than other patients. Further studies should focus on the collection of longitudinal patient data for more comprehensive insight

Pregnancy rates of day 4 and day 5 embryos after culture in an integrated time-lapse incubator

Background: The aim of this study was to compare pregnancy rates in patients undergoing IVF/ICSI with embryo transfer after 4 and 5 days of culture in a closed incubation system with integrated time-lapse imaging. Methods: Out of n = 2207 in vitro fertilization (IVF)/ intracytoplasmic sperm injection (ICSI) cycles performed between January 2011 and April 2016 at a tertiary referral university hospital, a total of n = 599 IVF/ICSI cycles with prolonged embryo culture in an integrated time-lapse system (EmbryoScope© (Vitrolife)) until day 4 or 5 were retrospectively analyzed with regard to embryo morphology and pregnancy rates. Results: A transfer on day 5 compared to a transfer on day 4 did not result in higher implantation and clinical pregnancy rates (IR 29.4% on day 4 versus 33.0% on day 5, p = 0.310; CPR 45.2% on day 4 versus 45.7% on day 5, p = 1.0). The percentage of ideal embryos transferred on day 4 was comparable to the rate of ideal embryos transferred on day 5 (41.6% versus 44.1%, p = 0.508). However, on day 4 a significantly higher number of embryos was transferred (1.92 on day 4 versus 1.84 on day 5, p = 0.023), which did not result in higher rates of multiple pregnancies. Conclusions: Pregnancy rates in IVF/ICSI cycles with integrated time-lapse incubation and transfer on day 4 and 5 are comparable. This finding provides the clinician, IVF laboratory and patient with more flexibility. Trial registration: This study was retrospectively registered by the local ethics committee of the University of Heidelberg on December 19, 2016 (registration number S-649/2016)

Structural impairment and conflict load as vulnerability factors for burnout – A cross-sectional study from the German working population

IntroductionIndividual vulnerability and resilience factors are increasingly studied in burnout research. This is especially true for clinical variables that translate directly into intervention programs from a psychodynamic perspective. For example, few studies have examined the relationship between structural impairment and the individual spectrum of motivational conflicts according to the Operationalized Psychodynamic Diagnosis system (OPD) in relation to burnout. To substantiate previous findings, we hypothesized that structural impairment as well as motivational conflicts are related to burnout, but that structural impairment explained additional variance and mediated a possible relationship between conflicts and burnout.MethodThe present cross-sectional study was carried out on a sample of the German working population (N = 545). Questionnaires were used to measure structural impairment (OPD-SQS), the conflict-modes along with the category K0 (OPD-CQ), as well as burnout (BOSS-I/-II).ResultsStructural impairment, a number of conflict modes, and burnout were significantly associated. Moreover, structural impairment explained additional variance in burnout. The requirements for the conflict-specific mediation models were given for 9 of the 12 OPD conflict modes. In these models the impact of the conflict modes on burnout was mediated by structural impairment.DiscussionThe current study broadens the comprehension of the relations between structural impairment, the conflict modes and burnout. In addition it higlights the role of structural impairment in predicting burnout risk and possible prevention approaches

Study protocol for a randomised controlled trial of an e-health stepped care approach for the treatment of internet use disorders versus a placebo condition: the SCAPIT study

Introduction Excessive internet use can lead to problems for some individuals. The WHO has introduced Gaming Disorder in the International Classification of Diseases-11 (ICD-11). Previous research has shown that other internet applications can cause serious mental health problems as well. It is important to provide measures of prevention, early intervention and therapy for internet use disorders (IUDs). Methods and analysis The study ‘Stepped Care Approach for Problematic Internet use Treatment’ is a randomised, two-arm, parallel-group, observer-blind trial. The aim of the study is to investigate if a stepped care approach is effective to reduce symptom severity for IUD. The sample is primarily recruited online with a focus on employees in companies with support of health insurances. After screening, the stepped care approach depends on the success of the previous step—that is, the successful reduction of criteria—and comprise: (1) app-intervention with questionnaires and feedback, (2) two telephone counsellings (duration: 50 min) based on motivational interviewing, (3) online therapy over 17 weeks (15 weekly group sessions, eight individual sessions) based on cognitive–behavioural therapy. A follow-up is conducted after 6 months. A total of 860 participants will be randomised. Hierarchical testing procedure is used to test the coprimary endpoints number of Diagnostic and Statistical Manual of Mental Disorders, fifth edition and ICD-11 criteria. Primary analysis will be performed with a sequential logit model. Ethics and dissemination The study has been approved by the Ethics Committees of the Universities of Lübeck (file number: 21-068), Mainz (file number: 2021-15907) and Berlin (file number: 015.2021). Results will be reported in accordance to the CONSORT statement. If the approach is superior to the control condition, it may serve as part of treatment for IUD. Trial registration number DRKS00025994

Технология сборки и дуговой сварки пластин из титанового сплава

Цель работы – является разработка технологии дуговой сварки пластин из сплава ВТ5-1. В процессе исследования проводился анализ повышения эффективности и сравнение способов сварки, ручной аргонодуговой сварки неплавящимся электродом и механизированной сварки плавящимся электродом в среде защитного газа. В результате работы были рассчитаны параметры и выбран наиболее производительный способ для сварки сплава ВТ5-1.The purpose of the work is to develop a technology for arc welding of plates made of VT5-1 alloy. In the course of the study, the analysis of efficiency improvement and comparison of welding methods, manual argon arc welding with a non-melting electrode and mechanized welding with a melting electrode in a protective gas environment were carried out. As a result of the work, the parameters were calculated and the most productive method for welding the VT5-1 alloy was selected

Heterophilic Binding of L1 on Unmyelinated Sensory Axons Mediates Schwann Cell Adhesion and Is Required for Axonal Survival

This study investigated the function of the adhesion molecule L1 in unmyelinated fibers of the peripheral nervous system (PNS) by analysis of L1- deficient mice. We demonstrate that L1 is present on axons and Schwann cells of sensory unmyelinated fibers, but only on Schwann cells of sympathetic unmyelinated fibers. In L1-deficient sensory nerves, Schwann cells formed but failed to retain normal axonal ensheathment. L1-deficient mice had reduced sensory function and loss of unmyelinated axons, while sympathetic unmyelinated axons appeared normal. In nerve transplant studies, loss of axonal-L1, but not Schwann cell-L1, reproduced the L1-deficient phenotype. These data establish that heterophilic axonal-L1 interactions mediate adhesion between unmyelinated sensory axons and Schwann cells, stabilize the polarization of Schwann cell surface membranes, and mediate a trophic effect that assures axonal survival

The PHENIX Experiment at RHIC

The physics emphases of the PHENIX collaboration and the design and current status of the PHENIX detector are discussed. The plan of the collaboration for making the most effective use of the available luminosity in the first years of RHIC operation is also presented.Comment: 5 pages, 1 figure. Further details of the PHENIX physics program available at http://www.rhic.bnl.gov/phenix