Thrombin-Derived C-Terminal Peptide Reduces Candida-Induced Inflammation and Infection In Vitro and In Vivo.

Infections due to the opportunistic fungus Candida have been on the rise in the last decades, especially in immunocompromised individuals and hospital settings. Unfortunately, the treatments available today are limited. Thrombin-derived C-terminal peptide (TCP-25) is an antimicrobial peptide (AMP) with antibacterial and immunomodulatory effects. In this work, we, for the first time, demonstrate the ability of TCP-25 ability to counteract Candida in vitro and in vivo. Using a combination of viable count assay (VCA), radial diffusion assay (RDA), and fluorescence and transmission electron microscopy analyses, TCP-25 was found to exert a direct fungicidal activity. An inhibitory activity of TCP-25 on NF-κB activation induced by both zymosan alone and heat-killed C. albicans was demonstrated in vitro using THP-1 cells, and in vivo using NF-κB reporter mice. Moreover, the immunomodulatory property of TCP-25 was further substantiated in vitro by analyzing cytokine responses in human blood stimulated with zymosan, and in vivo employing a zymosan-induced peritonitis model in C57BL/6 mice. The therapeutic potential of TCP-25 was demonstrated in mice infected with luminescent C. albicans. Finally, the binding between TCP-25 and zymosan was investigated using circular dichroism spectroscopy and intrinsic fluorescence analysis. Taken together, our results show that TCP-25 has a dual function by inhibiting Candida as well as the associated zymosan-induced inflammation. The latter function is accompanied by a change in secondary structure upon binding to zymosan. TCP-25, therefore, shows promise as a novel drug candidate against Candida infections.

Exhaustion disorder and altered brain activity in frontal cortex detected with fNIRS.

Patients with stress-related Exhaustion Disorder (ED) have problems with memory and executive function. These problems have been associated with deviant activity in prefrontal cortex (PFC). We investigated cognitive performance and functional activity in the PFC during prolonged mental activity in patients with ED (n = 20, 16 women) with a mean duration since diagnosis of 46 ± 23 months in comparison to healthy individuals (n = 20, 12 women). A block of six neuropsychological tests was performed in a sequence that was repeated once. The brain imaging technique, functional near infrared spectroscopy (fNIRS) was used for all tests. There were no differences between the groups in terms of changes over time, i.e. difference between first and second test block. In the Stroop-Simon test, the controls showedhigher functional activity in the frontal cortex. In the left ventrolateral PFC, we observed an increased activity in controls in the incongruent compared to the congruent trials, whereas no changes were detected in the ED patient group. During processing speed tasks, only ED patients showed higher functional activity in right dorsolateral PFC. The ED patients reported lower subjective energy level and they also performed less well on a mental control task compared to healthy individuals. In conclusion, ED patients showed altered functional activity compared to controls, indicating that ED patients process information differently in the prefrontal cortex, but the functional activity did not change during the 2½ hr procedure, as revealed by the test-retest design. Lay summary In this paper we show that patient with exhaustion disorder have a reduced functional activity in the prefrontal cortex. This functional activity was not affected by 2.5 hours mental activity.

A novel care guide for personalised palliative care - a national initiative for improved quality of care.

BACKGROUND: Even when palliative care is an integrated part of the healthcare system, the quality is still substandard for many patients and often initiated too late. There is a lack of structured guidelines for identifying and caring for patients; in particular for those with early palliative care needs. A care guide can act as a compass for best practice and support the care of patients throughout their palliative trajectory. Such a guide should both meet the needs of health care professionals and patients and families, facilitating discussion around end-of-life decision-making and enabling them to plan for the remaining time in life. The aim of this article is to describe the development and pilot testing of a novel Swedish palliative care guide. METHODS: The Swedish Palliative Care Guide (S-PCG) was developed according to the Medical Research Council framework and based on national and international guidelines for good palliative care. An interdisciplinary national advisory committee of over 90 health care professionals together with patient, family and public representatives were engaged in the process. The feasibility was tested in three pilot studies in different care settings. RESULTS: After extensive multi-unit and interprofessional testing and evaluation, the S-PCG contains three parts that can be used independently to identify, assess, address, follow up, and document the individual symptoms and care-needs throughout the whole palliative care trajectory. The S-PCG can provide a comprehensive overview and shared understanding of the patients' needs and possibilities for ensuring optimal quality of life, the family included. CONCLUSIONS: Based on broad professional cooperation, patients and family participation and clinical testing, the S-PCG provides unique interprofessional guidance for assessment and holistic care of patients with palliative care needs, promotes support to the family, and when properly used supports high-quality personalised palliative care throughout the palliative trajectory. Future steps for the S-PCG, entails scientific evaluation of the clinical impact and effect of S-PCG in different care settings - including implementation, patient and family outcomes, and experiences of patient, family and personnel.

In-vehicle fragrance administration as a countermeasure for driver fatigue.

Driver fatigue is a contributing factor in about 10-30% of all fatal crashes. Prevention of fatigue-related crashes relies on robust detection of driver fatigue and application of effective countermeasures. A potential countermeasure is fragrance administration since odors can have alerting effects on humans. The aim here was to investigate if a fragrance incorporating trigeminal components could be used as an in-vehicle countermeasure for driver fatigue. The fragrance was tested in a driving simulator with 21 healthy but sleep-deprived participants. Each participant performed a monotonous driving task twice, once with active fragrance containing a trigeminal component and once with olfactory fragrance, in a cross-over single-blind design. The order of trigeminal/olfactory fragrance was randomized and blinded to the participants. Both fragrances (trigeminal/olfactory) were administered either when the participant fell asleep (defined as eye closure > 3 s) or after approximately 45 min if the participant did not fall asleep. Self-reported sleepiness was assessed using the Karolinska Sleepiness Scale (KSS) every 5 min during driving. Variability in speed and lateral position and line crossing frequency were logged for each drive to measure driving performance. Heart rate measurements (ECG) and eye blinks (EOG) were collected to investigate potential arousing effects of the fragrance and to track objective signs of sleepiness. Mean blink duration, which was used as an objective measure of sleepiness, decreased significantly, after fragrance exposure, as did the frequency of line crossings, but there were no statistically significant differences between the fragrance with trigeminal stimulus and the pure olfactory fragrance. The results are in line with the effects found for other commonly used fatigue countermeasures, like playing loud music. These countermeasures can restore alertness and driving performance for a short while. Whether this is sufficient to support driving performance until the driver can make a safe stop in real traffic remains a topic for future studies.

Seat belt usage in buses - An observation study of usage and travellers' perspectives.

This study aims to evaluate seat belt usage in buses and to understand travellers' incentives of seat belt usage. Methods used are observational studies (10 cities, with 328 bus observations), focus group discussion (7 groups with a total of 32 participants) and a web survey (n = 1737 respondents). The results show that the seat belt use among bus passengers can be improved especially in regional and commercial bus traffic. It is more common to buckle up on long trips than on short trips. However, even though observations show high usage during long trips, travellers report that they remove the seat belt after a while if they want to sleep or for comfort reasons. For the bus drivers it is not possible to control passengers' usage. Dirty seat belts and technical malfunction might deter some passengers from using them and therefore systematic cleaning and control of seats and belts are recommended. On short trips one reason for not using the belt is related to worries about getting stuck and not being ready to get off in time. In general, it is most important to increase the usage on high-speed roads (>60 km/h), in lower speed it might be more important to provide a seat for each passenger. Based on the results a list of recommendations is presented.

The Bispecific Tumor Antigen-Conditional 4-1BB x 5T4 Agonist, ALG.APV-527, Mediates Strong T-Cell Activation and Potent Antitumor Activity in Preclinical Studies.

4-1BB (CD137) is an activation-induced costimulatory receptor that regulates immune responses of activated CD8 T and natural killer cells, by enhancing proliferation, survival, cytolytic activity, and IFNγ production. The ability to induce potent antitumor activity by stimulating 4-1BB on tumor-specific cytotoxic T cells makes 4-1BB an attractive target for designing novel immuno-oncology therapeutics. To minimize systemic immune toxicities and enhance activity at the tumor site, we have developed a novel bispecific antibody that stimulates 4-1BB function when co-engaged with the tumor-associated antigen 5T4. ALG.APV-527 was built on the basis of the ADAPTIR bispecific platform with optimized binding domains to 4-1BB and 5T4 originating from the ALLIGATOR-GOLD human single-chain variable fragment library. The epitope of ALG.APV-527 was determined to be located at domain 1 and 2 on 4-1BB using X-ray crystallography. As shown in reporter and primary cell assays in vitro, ALG.APV-527 triggers dose-dependent 4-1BB activity mediated only by 5T4 crosslinking. In vivo, ALG.APV-527 demonstrates robust antitumor responses, by inhibiting growth of established tumors expressing human 5T4 followed by a long-lasting memory immune response. ALG.APV-527 has an antibody-like half-life in cynomolgus macaques and was well tolerated at 50.5 mg/kg. ALG.APV-527 is uniquely designed for 5T4-conditional 4-1BB-mediated antitumor activity with potential to minimize systemic immune activation and hepatotoxicity while providing efficacious tumor-specific responses in a range of 5T4-expressing tumor indications as shown by robust activity in preclinical in vitro and in vivo models. On the basis of the combined preclinical dataset, ALG.APV-527 has potential as a promising anticancer therapeutic for the treatment of 5T4-expressing tumors.

Seroprevalence of SARS-CoV-2 antibodies among public transport workers in Sweden.

Introduction: Public transportation is an essential societal function in crisis situations like the coronavirus disease 2019 (COVID-19) pandemic. Bus drivers and other public transport workers are essential workers that need to keep working despite the risk of contagion. The SARS-CoV-2 virus may pose an occupational health risk to public transport workers and especially to bus drivers as they interact with passengers in a confined area. By analyzing antibodies towards SARS-CoV-2 proteins in blood samples it is possible to measure if an individual has been infected by COVID-19. Here, we report the prevalence of antibodies among bus drivers and other public transport employees in Stockholm, Sweden and relate it to socio-demographic factors. Methods: Seroprevalence of IgG antibodies towards SARS-CoV-2 proteins was investigated in a sample of 262 non-vaccinated public transport workers (182 men and 40 women) recruited between April 26 and May 7, 2021. Most of the participants were bus drivers (n = 222). The relationship between socio-demographic factors and seroprevalence was investigated with logistic regression. Results: The seroprevalence was 50% in the total sample of public transport workers. Among bus drivers, 51% were seropositive compared to 44% seropositive among the other public transport workers. The difference was not significant. The seroprevalence was higher than the national seroprevalence in Sweden during the same period (18.3% in non-vaccinated people aged 20-64 years). The logistic regression model using Wald forward selection showed that men had a higher risk of being seropositive (OR 2.7, 95% CI 1.3 - 5.8) and there was a higher risk with increasing number of people in the household (OR 1.3, 95% CI 1.1 - 1.6). Conclusions: These findings could imply an occupational risk for COVID-19 infection among public transport workers. Infection control measures are warranted during virus epidemics to assure bus drives' safety and reduce transmission in public transport.

Detecting driver fatigue using heart rate variability: A systematic review.

Driver fatigue detection systems have potential to improve road safety by preventing crashes and saving lives. Conventional driver monitoring systems based on driving performance and facial features may be challenged by the application of automated driving systems. This limitation could potentially be overcome by monitoring systems based on physiological measurements. Heart rate variability (HRV) is a physiological marker of interest for detecting driver fatigue that can be measured during real life driving. This systematic review investigates the relationship between HRV measures and driver fatigue, as well as the performance of HRV based fatigue detection systems. With the applied eligibility criteria, 18 articles were identified in this review. Inconsistent results can be found within the studies that investigated differences of HRV measures between alert and fatigued drivers. For studies that developed HRV based fatigue detection systems, the detection performance showed a large variation, where the detection accuracy ranged from 44% to 100%. The inconsistency and variation of the results can be caused by differences in several key aspects in the study designs. Progress in this field is needed to determine the relationship between HRV and different fatigue causal factors and its connection to driver performance. To be deployed, HRV-based fatigue detection systems need to be thoroughly tested in real life conditions with good coverage of relevant driving scenarios and a sufficient number of participants.

External communication of automated shuttles: Results, experiences, and lessons learned from three European long-term research projects.

Automated shuttles are already seeing deployment in many places across the world and have the potential to transform public mobility to be safer and more accessible. During the current transition phase from fully manual vehicles toward higher degrees of automation and resulting mixed traffic, there is a heightened need for additional communication or external indicators to comprehend automated vehicle actions for other road users. In this work, we present and discuss the results from seven studies (three preparatory and four main studies) conducted in three European countries aimed at investigating and providing a variety of such external communication solutions to facilitate the exchange of information between automated shuttles and other motorized and non-motorized road users.

LRIG1 and the liar paradox in prostate cancer: a study of the expression and clinical significance of LRIG1 in prostate cancer.

The course of prostate cancer varies greatly, and additional prognostic markers are needed. Leucine-rich repeats and immunoglobulin-like domains protein 1 (LRIG1) is an endogenous inhibitor of growth factor signaling and a proposed tumor suppressor. Publicly available gene expression datasets indicate that LRIG1 may be overexpressed in prostate cancer. In our study, the expression of LRIG1 protein in prostate cancer was evaluated for the first time. Immunohistochemistry was performed on tissue microarrays from two different patient series: 355 Swedish patients diagnosed by transurethral resection and 293 American patients who underwent radical prostatectomy. In the Swedish series, high expression of LRIG1 correlated with Gleason score, T-stage, tumor cell proliferation, vascular density and epidermal growth factor receptor (EGFR) phosphorylation. Among the 256 Swedish patients, followed by watchful waiting, high LRIG1 expression was significantly associated with short overall and prostate cancer-specific survival. In contrast, in the US series, high LRIG1 expression was significantly associated with long overall survival. In vitro cell experiments showed that LRIG1 was induced by androgen stimulation, and its expression inhibited prostate cancer cell proliferation. Thus, LRIG1 expression was an independent marker for poor survival in the untreated patient series, perhaps as a secondary marker of androgen receptor and/or EGFR activation. On the contrary, LRIG1 was a marker for good prognosis after prostatectomy, which might be due to its growth inhibiting properties. We propose that LRIG1 is an important determinant of prostate cancer growth, and the implications of its expression on patient outcome depend on the clinical and biological circumstances.

Evaluation of the prognostic significance of MSMB and CRISP3 in prostate cancer using automated image analysis.

Despite prostate cancer being the most frequent cancer in men in the Western world, tissue biomarkers for predicting disease recurrence after surgery have not been incorporated into clinical practice. Our group has previously identified ß-microseminoprotein (MSMB) and cysteine-rich secretory protein-3 (CRISP3) as independent predictors of biochemical recurrence after radical prostatectomy. The purpose of the present study was to use automated image analysis, enabling quantitative determination of MSMB and CRISP3 expressions in a large cohort and to validate the previous findings. MSMB and CRISP3 protein expressions were assessed on tissue microarrays constructed from 3268 radical prostatectomy specimens. Whole-slide digital images were captured, and a novel cytoplasmic algorithm was used to develop a quantitative scoring model for cytoplasmic staining. Classification regression tree analysis was used to group patients, with different risk for biochemical recurrence, depending on level of protein expression. Patients with tumors expressing high levels of MSMB had a significantly reduced risk for biochemical recurrence after radical prostatectomy (HR=0.468; 95% CI 0.394-0.556; P<0.001). Multivariate analysis adjusted for clinicopathological parameters revealed that MSMB expression was an independent predictor of decreased risk of recurrence (HR=0.710; 95% CI 0.578-0.872; P<0.001). We found no correlation between CRISP3 expression and biochemical recurrence. In this current study, we applied a novel image analysis on a large independent cohort and successfully verified that MSMB is a strong independent factor, predicting favorable outcome after radical prostatectomy for localized prostate cancer.

Cysteine-rich secretory protein 3 and ß-microseminoprotein on prostate cancer needle biopsies do not have predictive value for subsequent prostatectomy outcome.

OBJECTIVES: • To investigate whether cysteine-rich secretory protein 3 (CRISP-3) and/or ß-microseminoprotein (ß-MSP) expression in diagnostic prostate needle biopsies have predictive value for prostate cancer (PC) on radical prostatecomy (RP). • To evaluate their potential clinical implementation in a preoperative setting. PATIENTS AND METHODS: • In total, 174 participants from the European Randomized Study of Screening for Prostate Cancer, Rotterdam section, treated by RP for PC were included in the present study. • CRISP-3 and ß-MSP immunohistochemistry was performed on corresponding diagnostic needle biopsies. • Outcome was correlated with clinicopathological parameters (prostate-specific-antigen, PSA; number of positive biopsies; Gleason score, GS; pT-stage; surgical margins at RP) and significant PC at RP (pT3/4, or GS > 6, or tumour volume ≥ 0.5 mL) in the total cohort (n= 174) and in a subgroup with low-risk features at biopsy (PSA ≤ 10 ng/ml, cT ≤ 2, PSA density <0.20 ng/mL/g, GS < 7 and ≤ 2 positive biopsy cores; n= 87). RESULTS: • ß-MSP and CRISP-3 expression in PC tissue was heterogeneous, with variable staining intensities occurring in the same tissue specimen. • High expression of ß-MSP significantly correlated with GS < 7 at RP; it was not a predictor for significant PC at RP neither in the total group (n= 174; odds ratio, OR, 0.319; 95% confidence interval, CI, 0.060-1.695; P= 0.180), nor in the low-risk group (n= 87; OR, 0.227; 95% CI, 0.040-1.274; P= 0.092). • CRISP-3 expression was not related to clinicopathological parameters, and did not predict significant PC at RP in the total group (n= 174; OR, 1.056; 95% CI, 0.438-2.545; P= 0.904) or the low-risk group (n= 87; OR, 1.856; 95% CI, 0.626-5.506; P= 0.265). CONCLUSIONS: • High ß-MSP expression correlated with low GS in subsequent RP specimens, supporting the view that ß-MSP exerts a tumour-suppressive effect. • No significant prognostic value of ß-MSP or CRISP-3 in prostate needle biopsies for significant PC at RP was found. • ß-MSP or CRISP-3 do not have additional value in the therapeutic stratification of patients with PC.

The hypothalamo-pituitary-adrenal axis and the autonomic nervous system in burnout.

Burnout constitutes a serious health concern in the modern working environment. It is a stress-related condition that has developed as a result of a prolonged psychosocial stress exposure causing a persistent mismatch between demands and resources. The main symptom is emotional exhaustion, but physical fatigue, diminished professional efficacy, cynicism, and cognitive impairments are also associated with this condition. Burnout has been used both as a psychologic term in occupational settings and as a clinical diagnosis in patient populations, and there is currently no universally accepted definition and diagnostic criteria of burnout. It has been hypothesized that the two main stress response systems, the autonomic nervous system (ANS) and the hypothalamus-pituitary-adrenal axis (HPA axis), are involved in the pathogenesis of burnout. A common hypothesis is that in the early stages of chronic stress, the HPA axis and sympathetic ANS activity tend to be higher, while it will decrease with a longer duration of chronic stress to ultimately reach a state of hypoactivity in clinical burnout. The current research in this field shows many contradictory results. Thus there is no compelling evidence of either ANS or HPA dysfunction in burnout. However, there is partial support for the hypothesis of HPA and sympathetic hyperactivity in early stages, and HPA hyporeactivity and low vagal activity in more severe burnout cases, but high-quality studies investigating the causal links are still lacking.

Physiological and Cognitive Performance in F-22 Pilots During Day and Night Flying.

BACKGROUND: Many workers routinely transition between day and night shiftsincluding pilots, where night flights are commonly considered more stressful. The physiological toll from this transition is not fully understood, though fatigue is a factor in many aviation accidents. This research investigated the changes in physiological markers of stress and cognitive performance as F-22 pilots transitioned from day flying to night flying.METHODS: There were 17 fully-qualified F-22 pilots who took part in a 2-wk data collection using salivary swabs, wrist-worn activity monitors, the National Aeronautics and Space Administration-Task Load Index (NASA-TLX) inventory, and a go/no-go (GNG) test.RESULTS: No differences were found in comparing day and night flying on the GNG reaction time/accuracy, NASA-TLX scores, or sleep quantity. Cortisol levels were significantly higher than civilian levels in all experimental conditions and control days. Participants had higher than predicted cortisol levels postflight in the day-flying condition and lower than predicted cortisol levels postflight in the night-flying condition, relative to levels from control day patterns. We also found smaller changes in cortisol (pre- to postflight) in the day-flying condition for those with more F-22 experience. Finally, we found a negative correlation between Perceived Stress Scale scores and age of pilots (r 0.72).DISCUSSION: We hypothesized that the night-flying environment would be more stressful, but our results disputed this claim. Our results suggest day flying elicits more of a stress response; however, a larger sample size is required to verify results. Preliminary findings of potential stress adaptation may suggest stress adaptation in the F-22 community needs further investigation.Combs EK, Dahlman AS, Shattuck NL, Heissel JA, Whitaker LR. Physiological and cognitive performance in F-22 pilots during day and night flying. Aerosp Med Hum Perform. 2021; 92(5):303311.

The effects of exercise training on hypothalamic-pituitary-adrenal axis reactivity and autonomic response to acute stress-a randomized controlled study.

BACKGROUND: Exercise training is suggested to have a stress-buffering effect on physiological reactions to acute stress. The so-called cross-stressor adaptation hypothesis is one of many theories behind the plausible effects, proposing that the attenuated physiological reaction seen in trained individuals in response to acute exercise is also seen when the individual is exposed to acute psychosocial stress. However, few randomized controlled trials (RCT) are available in this field. Therefore, the aim of the present trial was to study the effects of a 6-month aerobic exercise intervention on the physiological response to acute laboratory stress. METHODS: A two-armed RCT including untrained but healthy individuals aged 20-50 years was conducted. Assessments included a peak oxygen uptake test and a psychosocial stress test (the Trier Social Stress Test). A total of 88 participants went through both baseline and follow-up measures (48 in the intervention group and 40 in the control group) with a similar proportion of women and men (20 women and 28 men in the intervention group and 18 women and 22 men in the control group). Outcome measures were adrenocorticotrophic hormone, cortisol, systolic and diastolic blood pressure, and heart rate responses to acute psychosocial stress. RESULTS: Oxygen uptake and time-to-exhaustion increased significantly following the intervention, while a decrease was seen in the control group. The analyses showed attenuated responses to acute psychosocial stress for all variables in both groups at follow-up, with no differences between the groups. No correlation was seen between amount of exercise training and reactivity to the stress test. Despite the increased oxygen uptake in the intervention group, no differences were seen between the groups for any of the outcome variables at follow-up. CONCLUSIONS: In this study, the cross-stressor adaptation hypothesis could not be confirmed. Both groups showed decreased reactions indicating a habituation to the stress test. TRIAL REGISTRATION: ClinicalTrials.gov NCT02051127 . Registered on 31 January 2014-retrospectively registered.

MECHANISMS IN ENDOCRINOLOGY: Endocrine and immunological aspects of burnout: a narrative review.

Burnout has several different definitions, and attempts have been made to discriminate between burnout as a psychological construct and burnout as a clinical entity. A large body of research has focused on elucidating the biological link between stress exposure and burnout and/or finding a clinically usable biomarker for burnout. The objective of this narrative review is to summarize the main endocrine and immune findings in relation to burnout. The literature has primarily focused on dysregulation of the hypothalamus-pituitary-adrenal (HPA) axis. However, albeit the large body of studies, it cannot be concluded that clear effects are seen on HPA axis function in people with burnout. The HPA axis and anabolic acute reactivity to stress might be affected in clinical burnout. Plausible, effects of chronic stress might rather be seen when measuring responses to acute stress rather than resting state hormonal levels. Studies on other hormones, including thyroid hormones, prolactin and growth hormone in burnout subjects are inconclusive. It is important to note that this field is faced with many methodological challenges, one being the diurnal and pulsatile nature of many of the hormones of interest, including cortisol, which is not always considered. Another challenge is the heterogeneity regarding definitions and measurements of stress and burnout. Existing studies on burnout and immune function are heterogeneous regarding the results and no firm conclusion can be made if clinically relevant immune changes are present in burnout subjects. An overall conclusion is that existing research cannot confirm any homogenous reliable endocrinological or immunological changes related to burnout.

Growth factors and neurotrophins in patients with stress-related exhaustion disorder.

Growth factors, such as vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF), and neurotrophic factors, including brain-derived neurotophic factor (BDNF), have attracted attention in studies of the biological effects of long-term stress exposure due to their neuroprotective roles. This study investigated whether circulating levels of EGF, VEGF and BDNF were altered in individuals with stress-related exhaustion disorder. Forty patients diagnosed with exhaustion disorder and 40 healthy subjects (50% women) provided fasting blood samples for analysis of EGF, VEGF, and BDNF in plasma. We found significantly lower levels of EGF, VEGF, and BDNF in patients with ED compared to healthy controls. This pattern was seen in both male and female patients. Given the important roles of BDNF and VEGF for brain plasticity and neurogenesis, decreased levels after long-term stress exposure could indicate increased risk of neuronal damage and cognitive impairments in this patient group.

The CTLA-4 x OX40 bispecific antibody ATOR-1015 induces anti-tumor effects through tumor-directed immune activation.

BACKGROUND: The CTLA-4 blocking antibody ipilimumab has demonstrated substantial and durable effects in patients with melanoma. While CTLA-4 therapy, both as monotherapy and in combination with PD-1 targeting therapies, has great potential in many indications, the toxicities of the current treatment regimens may limit their use. Thus, there is a medical need for new CTLA-4 targeting therapies with improved benefit-risk profile. METHODS: ATOR-1015 is a human CTLA-4 x OX40 targeting IgG1 bispecific antibody generated by linking an optimized version of the Ig-like V-type domain of human CD86, a natural CTLA-4 ligand, to an agonistic OX40 antibody. In vitro evaluation of T-cell activation and T regulatory cell (Treg) depletion was performed using purified cells from healthy human donors or cell lines. In vivo anti-tumor responses were studied using human OX40 transgenic (knock-in) mice with established syngeneic tumors. Tumors and spleens from treated mice were analyzed for CD8+ T cell and Treg frequencies, T-cell activation markers and tumor localization using flow cytometry. RESULTS: ATOR-1015 induces T-cell activation and Treg depletion in vitro. Treatment with ATOR-1015 reduces tumor growth and improves survival in several syngeneic tumor models, including bladder, colon and pancreas cancer models. It is further demonstrated that ATOR-1015 induces tumor-specific and long-term immunological memory and enhances the response to PD-1 inhibition. Moreover, ATOR-1015 localizes to the tumor area where it reduces the frequency of Tregs and increases the number and activation of CD8+ T cells. CONCLUSIONS: By targeting CTLA-4 and OX40 simultaneously, ATOR-1015 is directed to the tumor area where it induces enhanced immune activation, and thus has the potential to be a next generation CTLA-4 targeting therapy with improved clinical efficacy and reduced toxicity. ATOR-1015 is also expected to act synergistically with anti-PD-1/PD-L1 therapy. The pre-clinical data support clinical development of ATOR-1015, and a first-in-human trial has started (NCT03782467).

Exercise training and physiological responses to acute stress: study protocol and methodological considerations of a randomised controlled trial.

BACKGROUND: This paper describes the protocol and methodological prerequisites for a randomised controlled exercise intervention. Selected baseline data from the study are also presented, demonstrating some methodological challenges related to exercise intervention trials. The aim of the trial was to study the effects of exercise training on physiological responses to acute psychosocial stress in untrained individuals. METHODS: Individuals with a low level of physical activity were invited to participate in an exercise intervention lasting for 6 months. A total of 119 participants were included and went through a peak oxygen uptake test and a psychosocial stress test at baseline. Adrenocorticotropic hormone (ACTH) and cortisol were measured in connection to the stress test to identify the physiological response. RESULTS: Almost 90% of the participants reported themselves as untrained, but results from the objectively measured oxygen uptake did not seem to correspond to the reported sedentary lifestyle. The primary outcome measures at baseline varied between individuals. The mean change from pre-test to peak value was 214% for ACTH and 94% for cortisol. Of these, 13 individuals did not respond in ACTH and/or and cortisol. DISCUSSION: Supposedly untrained individuals seeking participation in an exercise intervention might not be as untrained as they report, a methodological consideration of importance when evaluating the effects of training. Another important consideration is related to the primary outcome measure, which should be measurable and possible to affect. Absence of reaction at baseline means that changes can only be detected as an increased reaction.

Simultaneous Targeting of Two Distinct Epitopes on MET Effectively Inhibits MET- and HGF-Driven Tumor Growth by Multiple Mechanisms.

Increased MET activity is linked with poor prognosis and outcome in several human cancers currently lacking targeted therapies. Here, we report on the characterization of Sym015, an antibody mixture composed of two humanized IgG1 antibodies against nonoverlapping epitopes of MET. Sym015 was selected by high-throughput screening searching for antibody mixtures with superior growth-inhibitory activity against MET-dependent cell lines. Synergistic inhibitory activity of the antibodies comprising Sym015 was observed in several cancer cell lines harboring amplified MET locus and was confirmed in vivo Sym015 was found to exert its activity via multiple mechanisms. It disrupted interaction of MET with the HGF ligand and prompted activity-independent internalization and degradation of the receptor. In addition, Sym015 induced high levels of CDC and ADCC in vitro The importance of these effector functions was confirmed in vivo using an Fc-effector function-attenuated version of Sym015. The enhanced effect of the two antibodies in Sym015 on both MET degradation and CDC and ADCC is predicted to render Sym015 superior to single antibodies targeting MET. Our results demonstrate strong potential for use of Sym015 as a therapeutic antibody mixture for treatment of MET-driven tumors. Sym015 is currently being tested in a phase I dose escalation clinical trial (NCT02648724). Mol Cancer Ther; 16(12); 2780-91. ©2017 AACR.