Sex-Specific associations of brain-derived neurotrophic factor and cardiorespiratory fitness in the general population

The brain-derived neurotrophic factor (BDNF) was initially considered to be neuron-specific. Meanwhile, this neurotrophin is peripherally also secreted by skeletal muscle cells and increases due to exercise. Whether BDNF is related to cardiorespiratory fitness (CRF) is currently unclear. We analyzed the association of serum BDNF levels with CRF in the general population (Study of Health in Pomerania (SHIP-TREND) from Northeast Germany; n = 1607, 51% female; median age 48 years). Sex-stratified linear regression models adjusted for age, height, smoking, body fat, lean mass, physical activity, and depression analyzed the association between BDNF and maximal oxygen consumption (VO2peak), maximal oxygen consumption normalized for body weight (VO2peak/kg), and oxygen consumption at the anaerobic threshold (VO2@AT). In women, 1mL/min higher VO2peak, VO2peak/kg, and VO2@AT were associated with a 2.43 pg/mL (95% confidence interval [CI]: 1.16 to 3.69 pg/mL; p = 0.0002), 150.66 pg/mL (95% CI: 63.42 to 237.90 pg/mL; p = 0.0007), and 2.68 pg/mL (95% CI: 0.5 to 4.8 pg/mL; p = 0.01) higher BDNF serum concentration, respectively. No significant associations were found in men. Further research is needed to understand the sex-specific association between CRF and BDNF. © 2019 by the authors. Licensee MDPI, Basel, Switzerland

Sex-Specific Associations of Brain-Derived Neurotrophic Factor and Cardiorespiratory Fitness in the General Population

The brain-derived neurotrophic factor (BDNF) was initially considered to be neuron-specific. Meanwhile, this neurotrophin is peripherally also secreted by skeletal muscle cells and increases due to exercise. Whether BDNF is related to cardiorespiratory fitness (CRF) is currently unclear. We analyzed the association of serum BDNF levels with CRF in the general population (Study of Health in Pomerania (SHIP-TREND) from Northeast Germany; n = 1607, 51% female; median age 48 years). Sex-stratified linear regression models adjusted for age, height, smoking, body fat, lean mass, physical activity, and depression analyzed the association between BDNF and maximal oxygen consumption (VO2peak), maximal oxygen consumption normalized for body weight (VO2peak/kg), and oxygen consumption at the anaerobic threshold (VO2@AT). In women, 1mL/min higher VO2peak, VO2peak/kg, and VO2@AT were associated with a 2.43 pg/mL (95% confidence interval [CI]: 1.16 to 3.69 pg/mL; p = 0.0002), 150.66 pg/mL (95% CI: 63.42 to 237.90 pg/mL; p = 0.0007), and 2.68 pg/mL (95% CI: 0.5 to 4.8 pg/mL; p = 0.01) higher BDNF serum concentration, respectively. No significant associations were found in men. Further research is needed to understand the sex-specific association between CRF and BDNF. © 2019 by the authors. Licensee MDPI, Basel, Switzerland

Mechanismen der Pathogen-Wirt-Interaktion – Störungen der Integrität des Zellverbandes humaner Atemwegsepithelzellen durch das α-Hämolysin von Staphylococcus aureus

Staphylococcus aureus, einer der häufigsten Erreger von Pneumonien, Endokardien und Sepsen (Frank et al. 2010), gehört bei nahezu einem Drittel der Bevölkerung zur normalen Nasenschleimhautflora (van Belkum et al. 2009) und kann unter bestimmten Risikobedingungen, vor allem in nosokomialer Umgebung, weiter in die unteren Atemwege vordringen und sich dort vermehren (van Belkum et al. 2009, Ahmed et al. 2015). Da das respiratorische Epithel von einer dicken, viskösen Mukusschicht bedeckt ist (Knowles & Boucher 2002), die Bakterien aufgrund ihrer Größe kaum durchdringen können, liegt die Hypothese nahe, dass es die sehr viel kleineren, löslichen Virulenzfaktoren der Bakterien sind, die den Mukus überqueren und einen ersten Pathogen-Wirt-Kontakt herstellen können. Das lösliche, porenbildende α-Hämolysin (Hämolysin a, Hla) ist einer der Haupt-Virulenzfaktor von S. aureus (Spaulding 2012). Studien hatten gezeigt, dass Hla auch in sublytischer Konzentration zu einer Auflösung der Zell-Zell- (Inoshima et al 2012) und Zell-Matrix-Kontakte (Hermann et al. 2015) humaner Atemwegsepithelzellen führte und so eine Lückenbildung im Zellverband induzierte. In vivo könnten solche Hla-vermittelten Prozesse dazu beitragen, dass eine erste Schädigung des Epithels erfolgt und die Überwindung der epithelialen Barriere für S. aures erleichtert wird. Die vorliegende Arbeit konnte in einem ersten Teil zeigen, dass diese Unfähigkeit von humanen Atemwegsepithelzellen (16HBE14o- und S9), nach Inkubation mit rHla den epithelialen Zusammenhalt aufrecht zu erhalten und entstandene parazelluläre Lücken durch aktive Migration zu schließen, auf eine rHla-induzierte Hyperphosphorylierung des fokalen Kontaktproteins Paxillin an Tyrosin 118 (und damit erhöhten Turnover der fokalen Kontakte) und Hypophosphorylierung des Actin-depolymerisierenden Faktors Cofilin an Serin 3 (und damit verstärkten Abbau von Stressfasern) zurückzuführen war. Der Hla-Effekt konnte so in fünf Prüfgrößen quantifizierbar erfasst werden: (1) Verlust des epithelialen Zusammenhalts, (2) Reorganisation des Actinzytoskeletts, (3) Auflösung fokaler Kontakte, (4) Hyperphosphorylierung von Paxillin und (5) Hypophosphorylierung von Cofilin. Im zweiten Teil der Arbeit wurden diese Prüfgrößen herangezogen, um den Mechanismus der Hla-Wirkung genauer aufzuklären. Durch Einsatz einer nichtporenbildenden Mutante rHla-H35L und dem Porenblocker IB201 konnte zunächst gezeigt werden, dass für die schädigenden Effekte auf den epithelialen Zusammenhalt der Zellen Ausbildung einer funktionellen Hla-Pore notwendig war und nicht Bindungsereignisse der Monomere, der Vorpore oder der Pore allein den Hla-Effekt auslösen konnten. Um die porenabhängigen Ereignisse zu untersuchen, wurden Ionenströme durch die Hla-Pore identifiziert und mit Ionomycin (erzeugt einen Calciumeinstrom) und Gramicidin (erzeugt einen Natriumeinstrom und Membrandepolarisierung) nachgebildet. Beide Ionenströme zusammen konnten den Hla-Effekt nahezu vollständig erzeugen. Die Ergebnisse wiesen darüber hinaus darauf hin, dass die Hla-erzeugten ionalen Veränderungen an der Membran unterschiedliche Signalveränderungen in der Zelle vermittelten: Calciumaktivierte Signalwege schienen vor allem für die beobachtete Paxillin-Phosphorylierung verantwortlich zu sein, während ein Natriumeinstrom zu einer Cofilin-Dephosphorylierung führte. Die genaue Signaltransduktion zwischen Einstrom der Ionen und (De-)Phosphorylierungsereignissen erfordert jedoch noch eine genauere Aufklärung. Des Weiteren konnte die Modellierung der Ionenströme den Hla-Effekt nicht komplett nachbilden, sodass wahrscheinlich zusätzliche porenabhängige Signalwege nach Hla-Behandlung (z.B. Verlust von ATP, Baaske & Richter et al. 2016) aktiviert werden.Staphyloccocus (S.) aureus is one of the most prominent pathogens leading to severe pneumonia, endocarditis and sepsis (Frank et al. 2010). Normally, colonizing the nares of about one third of the human population as a harmless commensal (van Belkum et al. 2009), S. aureus is able to penetrate and grow in the lower airways under certain risky condition, namely in nosocomial settings (van Belkum et al. 2009, Ahmed et al. 2015). The human respiratory epithelium in the lower airways is covered by a layer of thick, viscid mucus (Knowles and Boucher 2002), which bacteria are hardly able to pass even in situations of a high bacterial load in the lower airways. Therefore, it is more reasonable that much smaller, soluble virulence associated factors produced by S. aureus pass through the mucus and establish a first pathogen-host-interaction with the human airway epithelial cells. The soluble, poreforming α-hemolysin (hemolysin a, Hla) is one of the main virulence associated factors of S. aureus (Spaulding 2012). Previous studies have shown that sublytic concentrations of recombinant Hla (rHla) induces a dissociation of cell-to-cell (Inoshima et al. 2012) and cell-to-matrix contacts (Hermann et al. 2015) in human airway epithelial cells (16HBE14o- and S9), leading to a formation of paracellular gaps in the cell layer. In vivo, such Hla-mediated processes may initiate a first disturbance of the airway epithelial barrier and facilitate the entrance of S. aureus into the body. In the first part, the present study shows, that the inability of human airway epithelial cells to maintain the integrity of the cell layer and close paracellular gaps after treatment with Hla was due to Hla-induced hyper-phosphorylation of the focal contact protein paxillin at tyrosine 118 (leading to a higher turnover of focal contacts) and hypo-phosphorylation of the actin depolymerizing factor cofilin at serine 3 (leading to an increased depletion of stress fibers). For a quantifiable description of these Hla effects, five test categories were defined: (1) loss of the integrity of the cell layer, (2) reorganization of the actin cytoskeleton, (3) dissociation of focal contacts, (4) hyper-phosphorylation of paxillin, and (5) hypo-phosphorylation of cofilin. These test categories were subsequently used in the second part of the study to investigate the mechanism of action of Hla in more detail. First, using a non-poreforming mutant of Hla, Hla-H35L, and the pore blocking reagent IB201, it became evident that the formation of a functionally active Hla pore was a crucial requirement for Hla to elicit its harmful effects on the integrity of the cell layer of human airway epithelial cells. To study pore-dependent actions of Hla in more detail, possible ion currents were identified and modeled with ionomycin (inducing an influx of calcium) and gramicidin (inducing an influx of sodium and depolarizing the membrane potential). Both modeled ion currents combined were able to generate the effects of Hla almost entirely. Furthermore, the results indicated that these ionic changes mediated by Hla led to different signaling events in treated cells: Paxillin phosphorylation seemed to be mediated by activating calcium dependent signal pathways, whereas an influx of sodium ions resulted in cofilin dephosphorylation. Further investigation is required on how signaling transduction between an influx of ions and these (de)phosphorylation events occurs. In addition, modeling ion influxes did not entirely simulate the effects of Hla, indicating that additional pore-dependent events (i.e. the loss of ATP through the Hla pore, Baaske & Richter et al. 2016) probably account for the distinct effects of Hla

Structural and functional consequences in the amygdala of leptin-deficient mice

On the one hand, the emotional state can influence food intake and on the other hand, hunger can have an impact on the emotional state. Leptin, which is encoded by the ob gene, is involved in the energy homeostasis and plays a role in development of obesity. Mice deficient for leptin (ob/ob) are obese and display several behavioral alterations. It has been shown that ob/ob mice display striking changes in neuronal plasticity within the limbic system, e.g., hippocampal formation. We focus on alterations in ob/ob mice that can be related to alter processing in another part of the limbic system, the amygdala. ob/ob mice have a higher food consumption than age-matched controls, which might have an impact on the emotional state of these mice. Since the amygdala is involved in emotional processing, we analyze whether ob/ob mice display alterations in plasticity at the electrophysiological and structural level. No changes were seen in dendritic spine densities in the basolateral and lateral (LA) nucleus of the amygdala. Interestingly and in contrast to the hippocampus (Porter et al. 2013), long-term potentiation in the LA was increased in ob/ob mice. Our results indicate that amygdalar and hippocampal synaptic plasticity are regulated in different ways by leptin deficiency in accordance with the different functions of these limbic structures in stress and anxiety

ATP Release from Human Airway Epithelial Cells Exposed to Staphylococcus aureus Alpha-Toxin

Airway epithelial cells reduce cytosolic ATP content in response to treatment with S. aureus alpha-toxin (hemolysin A, Hla). This study was undertaken to investigate whether this is due to attenuated ATP generation or to release of ATP from the cytosol and extracellular ATP degradation by ecto-enzymes. Exposure of cells to rHla did result in mitochondrial calcium uptake and a moderate decline in mitochondrial membrane potential, indicating that ATP regeneration may have been attenuated. In addition, ATP may have left the cells through transmembrane pores formed by the toxin or through endogenous release channels (e.g., pannexins) activated by cellular stress imposed on the cells by toxin exposure. Exposure of cells to an alpha-toxin mutant (H35L), which attaches to the host cell membrane but does not form transmembrane pores, did not induce ATP release from the cells. The Hla-mediated ATP-release was completely blocked by IB201, a cyclodextrin-inhibitor of the alpha-toxin pore, but was not at all affected by inhibitors of pannexin channels. These results indicate that, while exposure of cells to rHla may somewhat reduce ATP production and cellular ATP content, a portion of the remaining ATP is released to the extracellular space and degraded by ecto-enzymes. The release of ATP from the cells may occur directly through the transmembrane pores formed by alpha-toxin