Balázs Radnai

TRAF6 is functional in inhibition of TLR4-mediated NF-κB activation by resveratrol☆

Resveratrol was suggested to inhibit Toll-like receptor (TLR)4-mediated activation of nuclear factor-κB (NF-κB) and Toll/interleukin-1 receptor domain-containing adaptor inducing interferon-β (TRIF)-(TANK)-binding kinase 1, but the myeloid differentiation primary response gene 88-tumor necrosis factor receptor-associated factor 6 (TRAF6) pathway is not involved in this effect. However, involvement of TRAF6 in this process is still elusive since cross talk between TRIF and TRAF6 has been reported in lipopolysaccharide (LPS)-induced signaling. Using RAW 264.7 macrophages, we determined the effect of resveratrol on LPS-induced TRAF6 expression, ubiquitination as well as activation of mitogen-activated protein (MAP) kinases and Akt in order to elucidate its involvement in TLR4 signaling. LPS-induced transient elevation in TRAF6 mRNA and protein expressions is suppressed by resveratrol. LPS induces the ubiquitination of TRAF6, which has been reported to be essential for Akt activation and for transforming growth factor-β activated kinase-1-NAP kinase kinase 6 (MKK6)-mediated p38 and c-Jun N-terminal kinase (JNK) activation. We found that resveratrol diminishes the effect of LPS on TRAF6 ubiquitination and activation of JNK and p38 MAP kinases, while it has no effect on the activation of extracellular-signal-regulated kinase (ERK)1/2. The effect of resveratrol on MAP kinase inhibition is significant since TRAF6 activation was reported to induce activation of JNK and p38 MAP kinase while not affecting ERK1/2. Moreover, Akt was identified previously as a direct target of TRAF6, and we found that, similarly to MAPKs, phosphorylation pattern of Akt followed the activation of TRAF6, and it was inhibited by resveratrol at all time points. Here, we provide the first evidence that resveratrol, by suppressing LPS-induced TRAF6 expression and ubiquitination, attenuates the LPS-induced TLR4-TRAF6, MAP kinase and Akt pathways that can be significant in its anti-inflammatory effects.

Suppressing LPS-induced early signal transduction in macrophages by a polyphenol degradation product: a critical role of MKP-1

Macrophages represent the first defense line against bacterial infection and therefore, play a crucial role in early inflammatory response. In this study, we investigated the role of MAPKs and MKP‐1 activation in regulation of an early inflammatory response in RAW 264.7 macrophage cells. We induced the inflammatory response by treating the macrophages with LPS and inhibited an early inflammatory response by using ferulaldehyde, a water‐soluble end‐product of dietary polyphenol degradation that we found previously to exert its beneficial anti‐inflammatory effects during the early phase of in vivo inflammation. We found that LPS‐induced ROS and nitrogen species formations were reduced by ferulaldehyde in a concentration‐dependent manner, and ferulaldehyde protected mitochondria against LPS‐induced rapid and massive membrane depolarization. LPS induced early suppression of MKP‐1, which was accompanied by activation of JNK, ERK, and p38 MAPK. By reversing LPS‐induced early suppression of MKP‐1, ferulaldehyde diminished MAPK activation, thereby inhibiting NF‐κB activation, mitochondrial depolarization, and ROS production. Taken together, our data suggest that ferulaldehyde exerts its early anti‐inflammatory effect by preserving the mitochondrial membrane integrity and shifting the expression of MKP‐1 forward in time in macrophages.

Ferulaldehyde, a Water-Soluble Degradation Product of Polyphenols, Inhibits the Lipopolysaccharide-Induced Inflammatory Response in Mice

Antiinflammatory properties of polyphenols in natural products, traditional medicines, and healthy foods were recently attributed to highly soluble metabolites produced by the microflora of the intestines rather than the polyphenols themselves. To provide experimental basis for this hypothesis, we measured antiinflammatory properties of ferulaldehyde (FA), a natural intermediate of polyphenol metabolism of intestinal microflora, in a murine lipopolysaccharide (LPS)-induced septic shock model. We found that intraperitoneally administered FA (6 mg/kg) prolonged the lifespan of LPS-treated (40 mg/kg) mice, decreased the inflammatory response detected by T(2)-weighted in vivo MRI, decreased early proinflammatory cytokines such as tumor necrosis factor-alpha and interleukin (IL)-1beta, and increased the antiinflammatory IL-10 in the sera of the mice. Additionally, FA inhibited LPS-induced activation of nuclear factor kappaB transcription factor in the liver of the mice. According to our data, these effects were probably due to attenuating LPS-induced activation of c-Jun N-terminal kinase and Akt. Furthermore, FA decreased free radical and nitrite production in LPS plus interferon-gamma-treated primary mouse hepatocytes, whose effects are expected to contribute to its antiinflammatory property. These data provide direct in vivo evidence, that a water-soluble degradation product of polyphenols could be responsible for, or at least could significantly contribute to, the beneficial antiinflammatory effects of polyphenol-containing healthy foods, natural products, and traditional medicines.

Protective effect of the poly(ADP-ribose) polymerase inhibitor PJ34 on mitochondrial depolarization-mediated cell death in hepatocellular carcinoma cells involves attenuation of c-Jun N-terminal kinase-2 and protein kinase B/Akt activation

Background2,4-Dimethoxyphenyl-E-4-arylidene-3-isochromanone (IK11) was previously described to induce apoptotic death of A431 tumor cells. In this report, we investigated the molecular action of IK11 in the HepG2 human hepatocellular carcinoma cell line to increase our knowledge of the role of poly (ADP-ribose)-polymerase (PARP), protein kinase B/Akt and mitogen activated protein kinase (MAPK) activation in the survival and death of tumor cells and to highlight the possible role of PARP-inhibitors in co-treatments with different cytotoxic agents in cancer therapy.ResultsWe found that sublethal concentrations of IK11 prevented proliferation, migration and entry of the cells into their G2 phase. At higher concentrations, IK11 induced reactive oxygen species (ROS) production, mitochondrial membrane depolarization, activation of c-Jun N-terminal kinase 2 (JNK2), and substantial loss of HepG2 cells. ROS production appeared marginal in mediating the cytotoxicity of IK11 since N-acetyl cysteine was unable to prevent it. However, the PARP inhibitor PJ34, although not a ROS scavenger, strongly inhibited both IK11-induced ROS production and cell death. JNK2 activation seemed to be a major mediator of the effect of IK11 since inhibition of JNK resulted in a substantial cytoprotection while inhibitors of the other kinases failed to do so. Inhibition of Akt slightly diminished the effect of IK11, while the JNK and Akt inhibitor and ROS scavenger trans-resveratrol completely protected against it.ConclusionsThese results indicate significant involvement of PARP, a marginal role of ROS and a pro-apoptotic role of Akt in this system, and raise attention to a novel mechanism that should be considered when cancer therapy is augmented with PARP-inhibition, namely the cytoprotection by inhibition of JNK2.

Synthesis and study of new paramagnetic resveratrol analogues

New resveratrol analogues containing five- and six-membered nitroxides and isoindoline nitroxides were synthesized. These new compounds were compared to resveratrol based on their ABTS radical scavenging ability as well on their capacity to suppress inflammatory process in macrophages induced by lipopolysaccharides. The ABTS and ROS scavenging activities of new molecules were the same or weaker than that of resveratrol, but some of paramagnetic resveratrol derivatives suppressed nitrite and TNFα production more efficiently than resveratrol. Based on these results the new nitroxide and phenol containing hybrid molecules can be considered as new antioxidant and anti-inflammatory agents.

Knowledge about basic life support in European students

INTRODUCTION Better knowledge and skills of basic life support can save millions of lives each year in Europe. AIM The aim of this study was to measure the knowledge about basic life support in European students. METHOD From 13 European countries 1527 volunteer participated in the survey. The questionnaire consisted of socio-demographic questions and knowledge regarding basic life support. The maximum possible score was 18. RESULTS Those participants who had basic life support training earned 11.91 points, while those who had not participated in lifesaving education had 9.6 points (p<0.001). Participants from former socialist Eastern European countries reached 10.13 points, while Western Europeans had average 10.85 points (p<0.001). The best results were detected among the Swedish students, and the worst among the Belgians. CONCLUSIONS Based on the results, there are significant differences in the knowledge about basic life support between students from different European countries. Western European youth, and those who were trained had better performance.

Little lifesavers: Can we start first aid education in kindergarten? – A longitudinal cohort study:

Objectives: First aid programmes should include not only the transfer of knowledge and skills but also improve participants’ helping attitudes. The aim of this study was to evalute the immediate and long-term effects of a 3-day first aid programme for kindergarten children. Methods: One hundred and eighteen kindergarten children (5–7 years old) from an inner-city kindergarten located in Pécs, Hungary, were involved in this study between January 2016 and June 2017. Training consisted of three 45-minute sessions involving the transfer of theoretical knowledge and practical skills about first aid. Knowledge, skills and attitudes were assessed by means of a questionnaire developed for the study and through observation. Participants were tested before and immediately after the programme and at 4 and 15 months. Results: The majority of children had no prior knowledge in first aid. Knowledge and skills significantly improved immediately after training and remained significantly higher at 4 and 15 months when compared to baseline – calling the ambulance (p < 0.01), performing cardiopulmonary resuscitation (CPR; p < .01), using an automated external defibrillator (AED; p < .01), handling an unconscious patient (p < .01) and managing bleeding (p < .01). Attitudes towards first aid improved significantly after the programme (p < .01) and remained improved after 4 and 15 months when compared to baseline (p < .01). Conclusion: Beginning first aid education in kindergarten can be useful. Children cannot become professional bystanders immediately following training, but the programme can offer an introduction of first aid. However, the results decreased substantially after 15 months so regular refreshing training should be recommended.

41 Is it necessary? – opinions and experiences about first aid education in childhood – pilot study

Aim Starting first aid education in early childhood can be an effective method of knowledge transfer. Our aim was to examine first aid education related opinions of kindergarten- and primary school children, educators, and parents. Method Eight-hundred seventy-one people (700 kindergarten- and primary school children, 29 educators, 142 parents) were involved in our study. Opinions were measured with self-edited questionnaires, containing mainly open questions. Results Based on the answers, the majority of children (90%) enjoyed our programme, they would like to participate further first aid programmes in the future. General opinions of parents and educators about first aid education in childhood were mainly positive, but they also gave negative ones. The majority of participants (>90%) had a positive opinion about our completed programme.1 Based on the opinions of parents and educators first aid education is recommended by health care professionals. Opinions were independent from gender (p=0.302), age (p=0.512), time of working with children (p=0.251). Conclusion Children’s opinion about the first aid programme were mainly positive. General opinions of parents and educators were different, but were positive related our programme. Based on this results opinions can change with concrete activities and can make first aid education more popular/acceptable by laypeople. Reference . Banfai B, Pek E, Pandur A, Csonka H, Betlehem J. ‘The year of first aid’: Effectiveness of a 3-day first aid programme for 7–14-year-old primary school children. Emerg Med J2017;34:526–32. Conflict of interest None Funding None

40 The long term effects of a 3-day first aid programme for 7–14 years old primary school children

Aim Bystanders can play an important role in the event of sudden injury or illness. Our aim was to evaluate the long term effects of a 3 day first aid programme for all primary school age groups (7–14 years old). Method This study was a 15 months follow-up of our previous investigation.1 Five-hundred twenty-four primary school children were involved in this study (288 girls and 236 boys). Measurements were made in the following topics: adult basic life support, using an automated external defibrillator, handling an unconscious patient, managing bleeding and calling the ambulance. Data collection was made with a self-made questionnaire and observation. Results Knowledge and skills after 15 months were significantly higher in the majority of the measured topics than before the programme (p<0.01). However, results were significantly worse than immediately and 4 months after programme (p<0.01). Younger children overall remembered less well than older children (p<0.01). Approximately three-third of children could use an AED, know the emergency phone number and the half of them could perform correct recovery position at 15 months. Correct assessment of breathing was similar than in baseline. Conclusion This study was the most comprehensive longitudinal study so far conducted on the effects of first aid (not only resuscitation) programme among primary school children. Participants could remember some aspects of first aid long term. However, forgetting after 15 months was significant so refreshing trainings would be mandatory annually Reference . Banfai B, Pek E, Pandur A, Csonka H, Betlehem J. ‘The year of first aid’: Effectiveness of a 3-day first aid programme for 7–14-year-old primary school children. Emerg Med J2017;34:526–32. Conflict of interest None Funding None

Munkahelyi stressz és megküzdési stratégiák vizsgálata földi és légi mentésben dolgozók körében

Absztrakt Bevezetes: A magyar egeszsegugyi szferaban dolgozok koreben koztudott a magas foku stresszhatas jelenlete, amely kihathat az egyenre. Celkitűzes: A foldi es legi mentesben dolgozok koreben feltarni a munkahelyi stresszt, annak merteket, illetve a pozitiv es negativ megkuzdesi strategiaikat. Modszer: 2015. juniustol oktoberig vizsgaltak Magyarorszagon a foldi es legi mentesben dolgozokat. Az adatgyűjtest sajat szerkesztesű, illetve a Rahe-fele, Roviditett Stressz- es Megkuzdesi Kerdőiv segitsegevel online formaban vegeztek. A kutatas keresztmetszeti tipusu, kvantitativ jellegű volt (n = 141). Az adatelemzes SPSS 20.0 statisztikai szoftverrel tortent. Szignifikanciahatar p<0,05 volt. Eredmenyek: Nagyobb a munkahelyi stresszhatas jelenlete a foldi mentesben dolgozoknal (p<0,01), nagyobb mertekben jelentkeznek naluk testi es pszichologiai tunetek (p<0,05). A Globalis Stressz es Megkuzdesi Index alapjan hatekonyabb copingmechanizmus figyelhető meg a legi mentesben dolgozoknal (p<0,01). Kovetkeztetese...