Interleukin-1 beta and neurotrophin-3 synergistically promote neurite growth in vitro

Pro-inflammatory cytokines such as interleukin-1 beta (IL-1β) are considered to exert detrimental effects during brain trauma and in neurodegenerative disorders. Consistently, it has been demonstrated that IL-1β suppresses neurotrophin-mediated neuronal cell survival rendering neurons vulnerable to degeneration. Since neurotrophins are also well known to strongly influence axonal plasticity, we investigated here whether IL-1β has a similar negative impact on neurite growth. We analyzed neurite density and length of organotypic brain and spinal cord slice cultures under the influence of the neurotrophins NGF, BDNF, NT-3 and NT-4. In brain slices, only NT-3 significantly promoted neurite density and length. Surprisingly, a similar increase of neurite growth was induced by IL-1β. Additionally, both factors increased the number of brain slices displaying maximal neurite growth. Furthermore, the co-administration of IL-1β and NT-3 significantly increased the number of brain slices displaying maximal neurite growth compared to single treatments. These data indicate that these two factors synergistically stimulate two distinct aspects of neurite outgrowth, namely neurite density and neurite length from acute organotypic brain slices

Mast cells protect from post-traumatic spinal cord damage in mice by degrading inflammation-associated cytokines via mouse mast cell protease 4

AbstractMast cells (MCs) are found abundantly in the central nervous system and play a complex role in neuroinflammatory diseases such as multiple sclerosis and stroke. In the present study, we show that MC-deficient KitW-sh/W-sh mice display significantly increased astrogliosis and T cell infiltration as well as significantly reduced functional recovery after spinal cord injury compared to wildtype mice. In addition, MC-deficient mice show significantly increased levels of MCP-1, TNF-α, IL-10 and IL-13 protein levels in the spinal cord. Mice deficient in mouse mast cell protease 4 (mMCP4), an MC-specific chymase, also showed increased MCP-1, IL-6 and IL-13 protein levels in spinal cord samples and a decreased functional outcome after spinal cord injury. A degradation assay using supernatant from MCs derived from either mMCP4−/− mice or controls revealed that mMCP4 cleaves MCP-1, IL-6, and IL-13 suggesting a protective role for MC proteases in neuroinflammation. These data show for the first time that MCs may be protective after spinal cord injury and that they may reduce CNS damage by degrading inflammation-associated cytokines via the MC-specific chymase mMCP4

FO‐SPR biosensor calibrated with recombinant extracellular vesicles enables specific and sensitive detection directly in complex matrices

Extracellular vesicles (EVs) have drawn huge attention for diagnosing myriad of diseases, including cancer. However, the EV detection and analyses procedures often lack much desired sample standardization. To address this, we used well-characterized recombinant EVs (rEVs) for the first time as a biological reference material in developing a fiber optic surface plasmon resonance (FO-SPR) bioassay. In this context, EV binding on the FO-SPR probes was achieved only with EV-specific antibodies (e.g. anti-CD9 and anti-CD63) but not with non-specific anti-IgG. To increase detection sensitivity, we tested six different combinations of EV-specific antibodies in a sandwich bioassay. Calibration curves were generated with two most effective combinations (anti-CD9/(B)anti-CD81 and anti-CD63/(B)anti-CD9), resulting in 10(3) and 10(4) times higher sensitivity than the EV concentration in human blood plasma from healthy or cancer patients, respectively. Additionally, by using anti-CD63/(B)anti-CD9, we detected rEVs spiked in cell culture medium and HEK293 endogenous EVs in the same matrix without any prior EV purification or enrichment. Lastly, we selectively captured breast cancer cell EVs spiked in blood plasma using anti-EpCA M antibody on the FO-SPR surface. The obtained results combined with FO-SPR real-time monitoring, fast response time and ease of operation, demonstrate its outstanding potential for EV quantification and analysis

Predicting bee community responses to land-use changes: Effects of geographic and taxonomic biases

Land-use change and intensification threaten bee populations worldwide, imperilling pollination services. Global models are needed to better characterise, project, and mitigate bees' responses to these human impacts. The available data are, however, geographically and taxonomically unrepresentative; most data are from North America and Western Europe, overrepresenting bumblebees and raising concerns that model results may not be generalizable to other regions and taxa. To assess whether the geographic and taxonomic biases of data could undermine effectiveness of models for conservation policy, we have collated from the published literature a global dataset of bee diversity at sites facing land-use change and intensification, and assess whether bee responses to these pressures vary across 11 regions (Western, Northern, Eastern and Southern Europe; North, Central and South America; Australia and New Zealand; South East Asia; Middle and Southern Africa) and between bumblebees and other bees. Our analyses highlight strong regionally-based responses of total abundance, species richness and Simpson's diversity to land use, caused by variation in the sensitivity of species and potentially in the nature of threats. These results suggest that global extrapolation of models based on geographically and taxonomically restricted data may underestimate the true uncertainty, increasing the risk of ecological surprises

Calibration of the CMS hadron calorimeters using proton-proton collision data at root s=13 TeV

Methods are presented for calibrating the hadron calorimeter system of theCMSetector at the LHC. The hadron calorimeters of the CMS experiment are sampling calorimeters of brass and scintillator, and are in the form of one central detector and two endcaps. These calorimeters cover pseudorapidities vertical bar eta vertical bar ee data. The energy scale of the outer calorimeters has been determined with test beam data and is confirmed through data with high transverse momentum jets. In this paper, we present the details of the calibration methods and accuracy.Peer reviewe

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Neuroimmune Communication in Skin: Far from Peripheral

The precise nature of the link between stress and exacerbation of skin inflammation has puzzled researchers and clinicians alike. The traditional explanation is that immune balance is altered by activation of two stress axes:activation of the hypothalamic–pituitary–adrenal (HPA) axis raises cortisol levels, and activation of the sympathetic nervous systems raises adrenaline levels (Figure 1). In this issue, Pavlovic et al. (2008) provide evidence for a third stress axis and report that stress travels to the skin through peripheral neuropeptidergic nerve fibers and exacerbates the neurogenic inflammatory aspect of cutaneous dermatitis. Clarification of the role of this alternative stress axis may enable the design of novel therapeutic strategie

Neuroimmunology of axonal regeneration

Titelblatt und Inhaltsverzeichnis Einleitung Zusammenfassung der eigenen Arbeiten Diskussion LiteraturverzeichnisIn der vorliegenden Habilitationsschrift konnte gezeigt werden, dass die Haut hochkontrolliert sich wiederholende Zyklen von Wachstum, Regression und Ruhe durchläuft (92, 93). Dieser physiologische Umbau geht einher mit dramatischen strukturellen Veränderungen, die alle Zelltypen und Kompartimente der Haut betreffen einschließlich Vaskularisierung (95), Innervation und Neurotrophin- Millieu (90, 91, 102, 112) sowie wichtige Aspekte des Immunsystems (87, 96, 103, 107, 108, 110, 111, 115). Sowohl die physiologische als auch die Entzündungs-assoziierte neuronale Plastizität der Haut werden wiederum stark durch psycho-emotionalen Stress beeinflusst (112). Wir konnten darüber hinaus zeigen, dass die pro-inflammatorischen Zytokine IL-1b, TNFa und IFNg sowie ein Cocktail dieser drei Zytokine die NGF-, NT-3-, NT-4 und p75NTR-Expression in epithelialen und mesenchymalen Zellpopulationen der Haut hochreguliert (85). Auf diesen Daten aufbauend untersuchten wir die Hypothese, ob die im Rahmen der Axotomie hochregulierten Zytokine IL-1b, IL-4, IL-6, TNFa und IFNg primär Neurotrophin-abhängiges Axonwachstum modulieren (86). Es zeigte sich, dass durch rekombinante Neurotrophine induziertes axonales Auswachsen durch alle untersuchten Zytokine außer durch IL-1b moduliert wurde. IL-6 und IFNg steigerten die Neurotrophin-abhängige Neuriten-Extension, während TNFa einen inhibierenden Einfluss hatte. IL-4 hatte einen dosisabhängigen Effekt: in niedriger Konzentration inhibierte und in hoher Konzentration förderte es NT-4-abhängiges Axonwachstum (86). Dies ist der erste Hinweis, dass das T -Zell-Zytokin IL-4 einen förderlichen Einfluss auf axonales Auswachsen hat. Schließlich konnten wir zeigen, dass T-Helfer-Zellen via IL-4 eine entscheidende Rolle bei der axonalen Regeneration nach mechanischer ZNS- Schädigung spielen (64). Aktivierte Th2-Zellen fördern über IL-4 signifikant das Neurotrophin-abhängige axonale Auswachsen nach entorhinaler Kortex-Läsion in vitro und in vivo sowie nach einer Kontusionsverletzung des Rückenmarks in Mäusen. Dies ist der erste formale Beleg, dass Th2-Zellen durch ihr Marker- Zytokin IL-4 Neurotrophin-abhängiges axonales Auswachsen nach ZNS-Läsion fördern (64). Die hier zusammengefassten Arbeiten sind ein klarer Hinweis darauf, dass die Zytokin/Neurotrophin-Achse in der neurowissenschaftlichen Forschung ein attraktives und neues pharmakologisches Ziel darstellt, um die Wundheilung und die axonale Regeneration nach traumatischer Schädigung des PNS und ZNS zu fördern.In the studies presented here, we were able to demonstrate that skin undergoes highly controlled cycles of growth, regression and rest. This physiological remodeling is associated with dramatic structural changes involving all cell types and compartments of skin including vascularization, innervation and neurotrophin milieu, as well as important aspects of the immune system. Additionally, physiological and inflammation-associated neuronal plasticity are substantially influenced by psycho-emotional stress. Furthermore, we demonstrated that proinflammatory cytokines such as IL-1-beta, TNF-alpha and IFN-gamma as well as a cocktail of these cytokines upregulate the expression of the neurotrophins NGF, NT-3, NT-4 and their receptor p75NTR in epithelial and mesenchymal cell populations of the skin. Based on these data, we tested the hypothesis that axotomy-induced upregulation of IL-1-beta, IL-4, IL-6, TNF-alpha and IFN-gamma modulate neurotrophin-dependent axon growth. We showed that axon outgrowth induced by recombinant neurotrophins is modulated by all factors investigated except IL-1-beta. IL-6 and IFN-gamma increase neurotrophin-dependent neurite extension, while TNF-alpha shows an inhibitory effect. IL-4 displays a dose-dependent effect: low concentrations inhibit and high concentrations stimulate NT-4-dependent axon growth. Finally, we demonstrated that T helper cells play a major role in axonal regeneration via IL-4 after traumatic CNS injury, such as entorhinal cortex lesion in vitro and in vivo, and contusion injury of the spinal cord in mice in vivo. This is the first formal proof that T helper cells stimulate neurotrophin-dependent axonal outgrowth after CNS injury via their marker cytokine IL-4. The studies summarized here indicate that the cytokine/neurotrophin axis is a promising, new pharmacological target in neuroscience research on wound healing and axonal outgrowth after traumatic injury of the PNS and CNS

Both Whistleblowers and the Scientists They Accuse Are Vulnerable and Deserve Protection

Whistleblowers play an important role diagnosing research misconduct, but often experience severe negative consequences. That is also true for incorrectly accused scientists. Both categories are vulnerable and deserve protection. Whistleblowers must proceed carefully and cautiously. Anonymous whistleblowing should be discouraged but cannot be ignored when the allegations are specific, serious, and plausible. When accused of a breach of research integrity it is important to be as transparent as possible. Sometimes accusations are false in the sense that the accuser knows or should know that the allegations are untrue. A mala fide whistleblower typically does not act carefully and we postulate a typology that may help in detecting them. Striking the right balance between whistleblower protection and timely unmasking false and identifying incorrect accusations is a tough dilemma leaders of research institutions have to face