Effects of ketamine, s-ketamine, and MK 801 on proliferation, apoptosis, and necrosis in pancreatic cancer cells.

BACKGROUND: Adenocarcinoma of the pancreas is one of the most aggressive cancer diseases affecting the human body. The oncogenic potential of this type of cancer is mainly characterized by its extreme growth rate triggered by the activation of signaling cascades. Modern oncological treatment strategies aim at efficiently modulating specific signaling and transcriptional pathways. Recently, anti-tumoral potential has been proven for several substances that are not primarily used in cancer treatment. In some tumor entities, for example, administration of glutamate antagonists inhibits cell proliferation, cell cycle arrest, and finally cell death. To attain endogenic proof of NMDA receptor type expression in the pancreatic cancer cell lines PaTu8988t and Panc-1 and to investigate the impact of ketamine, s-ketamine, and the NMDA receptor antagonist MK 801 on proliferation, apoptosis, and necrosis in pancreatic carcinoma. METHODS: Cell proliferation was measured by means of the ELISA BrdU assay, and the apoptosis rate was analyzed by annexin V staining. Immunoblotting were also used. RESULTS: The NMDA receptor type R2a was expressed in both pancreatic carcinoma cell lines. Furthermore, ketamine, s-ketamine, and MK 801 significantly inhibited proliferation and apoptosis. CONCLUSIONS: In this study, we showed the expression of the NMDA receptor type R2a in pancreatic cancer cells. The NMDA antagonists ketamine, s-ketamine, and MK 801 inhibited cell proliferation and cell death. Further clinical studies are warranted to identify the impact of these agents on the treatment of cancer patients.

Respiratory and Metabolic Responses of CD4 + T Cells to Acute Exercise and Their Association with Cardiorespiratory Fitness.

INTRODUCTION: The study aimed to investigate to what extent acute endurance exercise, especially eccentric exercise and cardiorespiratory fitness, affects the metabolic profile of CD4 + cells. METHODS: Fifteen male, healthy adults aged between 20 and 33 yr with a maximal oxygen uptake (V̇O 2max ) between 44 and 63 mL·kg -1 ·min -1 performed a downhill run (DR) and a level run (LR) for 45 min at 70% of their V̇O 2max on a treadmill in a crossover design. Blood samples were taken before (T0), directly after (T1), 3 h after (T3), and 24 h (T24) after each exercise for analyzing leukocyte numbers and cytokine levels. Isolated CD4 + cells were incubated for 4 h in autologous resting versus 3 h after exercise serum (T3 DR and T3 LR), and subsequently, cellular respiration, transcriptomic, and metabolomics profiles were measured. RESULTS: The systemic immune inflammation index increased significantly after DR and LR at T1 and T3 ( P < 0.001). In contrast, the transcriptomic and metabolic profile of CD4 + cells showed no significant alterations after incubation in T3 exercise serum. However, cardiorespiratory fitness positively correlated with the maximal mitochondrial respiration in CD4 + cells after incubation with T3 LR serum ( r = 0.617, P = 0.033) and with gene expression of oxidative phosphorylation and levels of different metabolites. Similarly, V̇O 2max was associated with an anti-inflammatory profile on RNA level. Lower lactate, methylmalonic acid, and d -gluconic acid levels were found in CD4 + cells of participants with a high V̇O 2max ( P < 0.001). CONCLUSIONS: Acute exercise leads to a mild proinflammatory milieu with only small changes in the metabolic homeostasis of CD4 + cells. High cardiorespiratory fitness is associated with a metabolic shift to oxidative phosphorylation in CD4 + cells. Functional relevance of this metabolic shift needs to be further investigated.

Exercise-induced inflammation alters the perception and visual exploration of emotional interactions.

Introduction: The study aimed to investigate whether an exercise-induced pro-inflammatory response alters the perception as well as visual exploration of emotional body language in social interactions. Methods: In a within-subject design, 19 male, healthy adults aged between 19 and 33 years performed a downhill run for 45 min at 70% of their VO2max on a treadmill to induce maximal myokine blood elevations, leading to a pro-inflammatory status. Two control conditions were selected: a control run with no decline and a rest condition without physical exercise. Blood samples were taken before (T0), directly after (T1), 3 h after (T3), and 24 h after (T24) each exercise for analyzing the inflammatory response. 3 h after exercise, participants observed point-light displays (PLDs) of human interactions portraying four emotions (happiness, affection, sadness, and anger). Participants categorized the emotional content, assessed the emotional intensity of the stimuli, and indicated their confidence in their ratings. Eye movements during the entire paradigm and self-reported current mood were also recorded. Results: The downhill exercise condition resulted in significant elevations of measured cytokines (IL6, CRP, MCP-1) and markers for muscle damage (Myoglobin) compared to the control running condition, indicating a pro-inflammatory state after the downhill run. Emotion recognition rates decreased significantly after the downhill run, whereas no such effect was observed after control running. Participants' sensitivity to emotion-specific cues also declined. However, the downhill run had no effect on the perceived emotional intensity or the subjective confidence in the given ratings. Visual scanning behavior was affected after the downhill run, with participants fixating more on sad stimuli, in contrast to the control conditions, where participants exhibited more fixations while observing happy stimuli. Conclusion: Our study demonstrates that inflammation, induced through a downhill running model, impairs perception and emotional recognition abilities. Specifically, inflammation leads to decreased recognition rates of emotional content of social interactions, attributable to diminished discrimination capabilities across all emotional categories. Additionally, we observed alterations in visual exploration behavior. This confirms that inflammation significantly affects an individual's responsiveness to social and affective stimuli.

T-cell-derived TNF-α and a cluster of immunological parameters from plasma allow a separation between SARS-CoV-2 convalescent versus vaccinated elite athletes.

Guidelines for medical clearing after SARS-CoV-2 infection in elite athletes do not include T-cell immunity aspects despite its relevance in the course of COVID-19 disease. Therefore, we aimed to analyze T-cell-related cytokines before and after in-vitro activation of CD4+ T-cells. We sampled professional indoor sports athletes at medical clearing after SARS-CoV-2 infection obtaining clinical, fitness data, and serological data including CD4+ T-cell cytokines. All data were analyzed by principal component analysis and 2 × 2 repeated measures ANOVA. CD4+ T-cells were sampled for cell culture activation with anti-CD3/anti-CD28 tetramers. At medical clearing, CD4+ T-cells from convalescent athletes secreted increased levels of TNF-α 72 h after in-vitro activation compared to vaccinated athletes. IL-18 levels in plasma were elevated and a cluster of parameters differentiated convalescent from vaccinated athletes by 13 parameters at the timepoint of medical clearing. All clinical data indicate infection is resolved, while increased TNF-α may reflect altered proportions of peripheral T-cells as a hangover of infection.

Recovery-Stress Response of Blood-Based Biomarkers.

The purpose of this study was to investigate blood-based biomarkers and their regulation with regard to different recovery-stress states. A total of 35 male elite athletes (13 badminton, 22 soccer players) were recruited, and two venous blood samples were taken: one in a 'recovered' state (REC) after a minimum of one-day rest from exercise and another one in a 'non-recovered' state (NOR) after a habitual loading microcycle. Overall, 23 blood-based biomarkers of different physiologic domains, which address inflammation, muscle damage, and tissue repair, were analyzed by Luminex assays. Across all athletes, only creatine kinase (CK), interleukin (IL-) 6, and IL-17A showed higher concentrations at NOR compared to REC time points. In badminton players, higher levels of CK and IL-17A at NOR were found. In contrast, a higher value for S100 calcium-binding protein A8 (S100A8) at REC was found in badminton players. Similar differences were found for BDNF in soccer players. Soccer players also showed increased levels of CK, and IL-6 at NOR compared to REC state. Several molecular markers were shown to be responsive to differing recovery-stress states, but their suitability as biomarkers in training must be further validated.

Abdominal Obesity-Related Disturbance of Insulin Sensitivity Is Associated with CD8+ EMRA Cells in the Elderly.

Aging and overweight increase the risk of developing type 2 diabetes mellitus. In this cross-sectional study, we aimed to investigate the potential mediating role of T-EMRA cells and inflammatory markers in the development of a decreased insulin sensitivity. A total of 134 healthy older volunteers were recruited (age 59.2 (SD 5.6) years). T cell subpopulations were analyzed by flow cytometry. Furthermore, body composition, HOMA-IR, plasma tryptophan (Trp) metabolites, as well as cytokines and adipokines were determined. Using subgroup and covariance analyses, the influence of BMI on the parameters was evaluated. Moreover, correlation, multiple regression, and mediation analyses were performed. In the subgroup of participants with obesity, an increased proportion of CD8+EMRA cells and elevated concentrations of plasma kynurenine (KYN) were found compared to the lower-weight subgroups. Linear regression analysis revealed that an elevated HOMA-IR could be predicted by a higher proportion of CD8+EMRA cells and KYN levels. A mediation analysis showed a robust indirect effect of the Waist-to-hip ratio on HOMA-IR mediated by CD8+EMRA cells. Thus, the deleterious effects of abdominal obesity on glucose metabolism might be mediated by CD8+EMRA cells in the elderly. Longitudinal studies should validate this assumption and analyze the suitability of CD8+EMRA cells as early predictors of incipient prediabetes.

Physical Activity and Mental Health of Patients with Pulmonary Hypertension during the COVID-19 Pandemic.

The aim of the study was to analyze the effect of personal restrictions on physical activity, mental health, stress experience, resilience, and sleep quality in patients with pulmonary hypertension (PH) during the "lockdown" period of the COVID-19 pandemic. In total, 112 PH patients and 52 age-matched healthy control subjects completed a questionnaire on the topics of physical activity, mental health, resilience, and sleep quality. PH patients had significantly lower physical activity, mental health, and sleep quality compared to age-matched healthy controls. Physical activity positively correlated with mental health and sleep quality in the PH group. Mental wellbeing and life satisfaction could be predicted by total physical activity, sleep, stress level, and resilience. PH patients appeared as an especially vulnerable group, demanding interventions to promote an active lifestyle and protect mental health in these patients. This could be helpful in counseling on how to carry out physical activity while maintaining infection control.

Neoadjuvant treatment of colorectal cancer with bevacizumab: the perioperative angiogenic balance is sensitive to systemic thrombospondin-1 levels.

PURPOSE: Colorectal cancer patients receiving neoadjuvant treatment with bevacizumab, a monoclonal antibody neutralizing vascular endothelial growth factor (VEGF), may suffer from wound healing complications after surgery as the antibody persists in patient blood. We characterized the systemic angiogenic balance in the perioperative period to evaluate its effect on physiologic angiogenesis. EXPERIMENTAL DESIGN: Nineteen patients receiving combination chemotherapy and bevacizumab for six neoadjuvant cycles were compared with 14 patients receiving chemotherapy without bevacizumab. Plasma from perioperative days -1, +1, +7, and +21 was analyzed for VEGF, thrombospondin-1 (TSP-1), and PD-ECGF concentrations. The angiogenic capacity was further tested in an in vitro assay of endothelial cell proliferation and migration. RESULTS: On day +1, the onset of wound healing was reflected in a change of balance, i.e., an increase of proangiogenic factors VEGF and platelet-derived endothelial cell growth factor compared with low TSP-1 inhibitor levels in both treatment groups. Patients with bevacizumab therapy showed significantly higher blood levels of total VEGF throughout the evaluation period. However, most VEGF molecules were inactive, i.e., complexed with antibody. Nevertheless, the capacity to stimulate endothelial growth was higher for these plasma samples and was reflected in low TSP-1 levels and an altered TSP-1 sensitivity. When purified TSP-1 protein was added, plasma samples of the bevacizumab but not the chemotherapy group showed reduced endothelial growth. CONCLUSIONS: Feedback mechanisms of bevacizumab therapy are not restricted to VEGF expression but seem to involve additional factors, such as TSP-1, which influences the systemic angiogenic balance and permits endothelial growth.

[Death after refusing treatment--causality assessment using the example of a Jehovah's Witness]. / Tod nach Therapieverweigerung. Eine Kausalitätsprüfung am Beispiel eines Zeugen Jehovas.

A 39-year-old man and Jehova's Witness suffered a complex pelvic fracture in an accident at work. He died 17 days later from fulminant pulmonary embolism. For religious reasons he had refused blood transfusions which would have been necessary for an early surgical stabilization of the pelvic fracture. Alternatively, the clinicians initiated a therapy with recombinant human erythropoetin (EPO) to increase haemoglobin and postponed the surgical stabilization of the pelvic fracture to a later date. Shortly before the planned operation, the patient suddenly died. The forensic autopsy confirmed the suspected diagnosis of pulmonary embolism as the cause of death; the manner of death was classified as an accident. Because of the preceding refusal of treatment the causality between the refused blood transfusion with the subsequent alternative therapy and the patient's death had to be assessed.

Point-of-care platelet function tests: detection of platelet inhibition induced by nonopioid analgesic drugs.

Detection of platelet inhibition is of clinical relevance in the preinterventional risk-benefit assessment in chronic low-back-pain patients scheduled for invasive pain therapy. We evaluated the sensitivity of various point-of-care platelet function tests for the detection of platelet inhibition induced by nonopioid analgesic drugs. After Institutional Review Board approval and informed consent, citrated whole blood from 40 patients with chronic unspecific low back pain was investigated before and 30 min after intravenous infusion of the study medication consisting of diclofenac 75 mg (plus orphenadrin 30 mg; Neodolpasse; Fresenius Kabi Austria GmbH, Austria), parecoxib 40 mg (Dynastat; Pharmacia Europe EEIG, UK), paracetamol 1 g (Perfalgan; Bieffe Medital S.P.A., Italy), or normal saline in a randomized, cross-over, double-blinded, placebo-controlled study. Platelet function was assessed using the PFA-100 platelet function analyzer and thromboelastometry, as well as impedance aggregometry (in the last 17 patients recruited after it became commercially available). Sensitivity for detecting diclofenac-induced platelet inhibition was 85% for the PFA-100 using epinephrine as agonist and 94% for arachidonic acid-induced impedance aggregometry. ADP-induced platelet function tests, as well as cytochalasin D-modified thromboelastometry were unreliable. All tests had a low incidence of false-positive test results after normal saline. Paracetamol and parecoxib had no significant platelet inhibiting effect. The PFA-100 using epinephrine as agonist and arachidonic acid-induced impedance aggregometry are recommended for the detection of cyclooxygenase-I-inhibiting effects of nonsteroidal anti-inflammatory drugs such as diclofenac. Our findings confirm that a single rescue dose of paracetamol and parecoxib has no antiplatelet effect.