β2-adrenergic agonists modulate TNF-α induced astrocytic inflammatory gene expression and brain inflammatory cell populations

Background: The NF-kappa B signaling pathway orchestrates many of the intricate aspects of neuroinflammation. Astrocytic beta(2)-adrenergic receptors have emerged as potential regulators in central nervous system inflammation and are potential targets for pharmacological modulation. The aim of this study was to elucidate the crosstalk between astrocytic beta(2)-adrenergic receptors and the TNF-alpha induced inflammatory gene program. Methods: Proinflammatory conditions were generated by the administration of TNF-alpha. Genes that are susceptible to astrocytic crosstalk between beta(2)-adrenergic receptors (stimulated by clenbuterol) and TNF-alpha were identified by qPCR-macroarray-based gene expression analysis in a human 1321 N1 astrocytoma cell line. Transcriptional patterns of the identified genes in vitro were validated by RT-PCR on the 1321 N1 cell line as well as on primary rat astrocytes. In vivo expression patterns were examined by intracerebroventricular administration of clenbuterol and/or TNF-alpha in rats. To examine the impact on the inflammatory cell content of the brain we performed extensive FACS analysis of rat brain immune cells after intracerebroventricular clenbuterol and/or TNF-alpha administration. Results: Parallel transcriptional patterns in vivo and in vitro confirmed the relevance of astrocytic beta(2)-adrenergic receptors as modulators of brain inflammatory responses. Importantly, we observed pronounced effects of beta(2)-adrenergic receptor agonists and TNF-alpha on IL-6, CXCL2, CXCL3, VCAM1, and ICAM1 expression, suggesting a role in inflammatory brain cell homeostasis. Extensive FACS-analysis of inflammatory cell content in the brain demonstrated that clenbuterol/TNF-alpha co-administration skewed the T cell population towards a double negative phenotype and induced a shift in the myeloid brain cell population towards a neutrophilic predominance. Conclusions: Our results show that astrocytic beta(2)-adrenergic receptors are potent regulators of astrocytic TNF-alpha-activated genes in vitro and in vivo, and ultimately modulate the molecular network involved in the homeostasis of inflammatory cells in the central nervous system. Astrocytic beta(2)-adrenergic receptors and their downstream signaling pathway may serve as potential targets to modulate neuroinflammatory responses

Change-point detection of peak tibial acceleration in overground running retraining

A method is presented for detecting changes in the axial peak tibial acceleration while adapting to self-discovered lower-impact running. Ten runners with high peak tibial acceleration were equipped with a wearable auditory biofeedback system. They ran on an athletic track without and with real-time auditory biofeedback at the instructed speed of 3.2 m·s−1. Because inter-subject variation may underline the importance of individualized retraining, a change-point analysis was used for each subject. The tuned change-point application detected major and subtle changes in the time series. No changes were found in the no-biofeedback condition. In the biofeedback condition, a first change in the axial peak tibial acceleration occurred on average after 309 running gait cycles (3′40″). The major change was a mean reduction of 2.45 g which occurred after 699 running gait cycles (8′04″) in this group. The time needed to achieve the major reduction varied considerably between subjects. Because of the individualized approach to gait retraining and its relatively quick response due to a strong sensorimotor coupling, we want to highlight the potential of a stand-alone biofeedback system that provides real-time, continuous, and auditory feedback in response to the axial peak tibial acceleration for lower-impact running

Tibial acceleration-based prediction of maximal vertical loading rate during overground running : a machine learning approach

Ground reaction forces are often used by sport scientists and clinicians to analyze the mechanical risk-factors of running related injuries or athletic performance during a running analysis. An interesting ground reaction force-derived variable to track is the maximal vertical instantaneous loading rate (VILR). This impact characteristic is traditionally derived from a fixed force platform, but wearable inertial sensors nowadays might approximate its magnitude while running outside the lab. The time-discrete axial peak tibial acceleration (APTA) has been proposed as a good surrogate that can be measured using wearable accelerometers in the field. This paper explores the hypothesis that applying machine learning to time continuous data (generated from bilateral tri-axial shin mounted accelerometers) would result in a more accurate estimation of the VILR. Therefore, the purpose of this study was to evaluate the performance of accelerometer-based predictions of the VILR with various machine learning models trained on data of 93 rearfoot runners. A subject-dependent gradient boosted regression trees (XGB) model provided the most accurate estimates (mean absolute error: 5.39 +/- 2.04 BW.s(-1), mean absolute percentage error: 6.08%). A similar subject-independent model had a mean absolute error of 12.41 +/- 7.90 BW.s(-1) (mean absolute percentage error: 11.09%). All of our models had a stronger correlation with the VILR than the APTA (p < 0.01), indicating that multiple 3D acceleration features in a learning setting showed the highest accuracy in predicting the lab-based impact loading compared to APTA

How the venom from the ectoparasitoid wasp Nasonia vitripennis exhibits anti-inflammatory properties on mammalian cell lines

With more than 150,000 species, parasitoids are a large group of hymenopteran insects that inject venom into and then lay their eggs in or on other insects, eventually killing the hosts. Their venoms have evolved into different mechanisms for manipulating host immunity, physiology and behavior in such a way that enhance development of the parasitoid young. The venom from the ectoparasitoid Nasonia vitripennis inhibits the immune system in its host organism in order to protect their offspring from elimination. Since the major innate immune pathways in insects, the Toll and Imd pathways, are homologous to the NF-kappa B pathway in mammals, we were interested in whether a similar immune suppression seen in insects could be elicited in a mammalian cell system. A well characterized NF-kappa B reporter gene assay in fibrosarcoma cells showed a dose-dependent inhibition of NF-kappa B signaling caused by the venom. In line with this NF-kappa B inhibitory action, N. vitripennis venom dampened the expression of IL-6, a prototypical proinflammatory cytokine, from LPS-treated macrophages. The venom also inhibited the expression of two NF-kappa B target genes, I kappa B alpha and A20, that act in a negative feedback loop to prevent excessive NF-kappa B activity. Surprisingly, we did not detect any effect of the venom on the early events in the canonical NF-kappa B activation pathway, leading to NF-kappa B nuclear translocation, which was unaltered in venom-treated cells. The MAP kinases ERK, p38 and JNK are other crucial regulators of immune responses. We observed that venom treatment did not affect p38 and ERK activation, but induced a prolonged JNK activation. In summary, our data indicate that venom from N. vitripennis inhibits NF-kappa B signaling in mammalian cells. We identify venom-induced up regulation of the glucocorticoid receptor-regulated GILZ as a most likely molecular mediator for this inhibition

Ubiquitin D regulates IRE1 α/c-Jun N-terminal kinase (JNK) protein-dependent apoptosis in pancreatic beta cells

Pro-inflammatory cytokines contribute to pancreatic beta cell apoptosis in type 1 diabetes at least in part by inducing endoplasmic reticulum (ER) stress and the consequent unfolded protein response (UPR). It remains to be determined what causes the transition from "physiological" to "apoptotic" UPR, but accumulating evidence indicates that signaling by the ER transmembrane protein IRE1 alpha is critical for this transition. IRE1 alpha activation is regulated by both intra-ER and cytosolic cues. We evaluated the role for the presently discovered cytokine-induced and IRE1 alpha-interacting protein ubiquitin D (UBD) on the regulation of IRE1 alpha and its downstream targets. UBD was identified by use of a MAPPIT (mammalian protein-protein interaction trap)-based IRE1 alpha interactome screen followed by comparison against functional genomic analysis of human and rodent beta cells exposed to pro-inflammatory cytokines. Knockdown ofUBDin human and rodent beta cells and detailed signal transduction studies indicated that UBD modulates cytokine-induced UPR/IRE1 alpha activation and apoptosis. UBD expression is induced by the pro-inflammatory cytokines interleukin (IL)-1 beta and interferon (IFN)-gamma in rat and human pancreatic beta cells, and it is also up-regulated in beta cells of inflamed islets from non-obese diabetic mice. UBD interacts with IRE1 alpha in human and rodent beta cells, modulating IRE1 alpha-dependent activation of JNK and cytokine-induced apoptosis. Our data suggest that UBD provides a negative feedback on cytokine-induced activation of the IRE1 alpha/JNK pro-apoptotic pathway in cytokine-exposed beta cells

Evaluation of an Immunoturbidimetric Assay for Haemoglobin A1c on a Cobas® Mira S Analyser

Peer Reviewe

Interleukin-1 as innate mediator of T cell immunity

The three-signal paradigm tries to capture how the innate immune system instructs adaptive immune responses in three well-defined actions: (1) presentation of antigenic peptides in the context of MHC molecules, which allows for a specific T cell response; (2) T cell co-stimulation, which breaks T cell tolerance; and (3) secretion of polarizing cytokines in the priming environment, thereby specializing T cell immunity. The three-signal model provides an empirical framework for innate instruction of adaptive immunity, but mainly discusses STAT-dependent cytokines in T cell activation and differentiation, while the multi-faceted roles of type I IFNs and IL-1 cytokine superfamily members are often neglected. IL-1 alpha and IL-1 beta are pro-inflammatory cytokines, produced following damage to the host (release of DAMPs) or upon innate recognition of PAMPs. IL-1 activity on both DCs and T cells can further shape the adaptive immune response with variable outcomes. IL-1 signaling in DCs promotes their ability to induce T cell activation, but also direct action of IL-1 on both CD4(+) and CD8(+) T cells, either alone or in synergy with prototypical polarizing cytokines, influences T cell differentiation under different conditions. The activities of IL-1 form a direct bridge between innate and adaptive immunity and could therefore be clinically translatable in the context of prophylactic and therapeutic strategies to empower the formation of T cell immunity. Understanding the modalities of IL-1 activity during T cell activation thus could hold major implications for rational development of the next generation of vaccine adjuvants

On the whereabouts of SARS-CoV-2 in the human body : a systematic review

Author summary Since the beginning of 2020, SARS-CoV-2 quickly spread throughout the human population and caused a pandemic with devastating consequences at a global scale. The scientific community is challenged to find good strategies for the containment and treatment of this virus. In this context, an important step is charting the viral presence in the human body to improve diagnostics, prevention or treatment. Here, we bring together the current scientific knowledge on SARS-CoV-2 detection in the human body and body fluids. We observe that SARS-CoV-2 impacts the human body well beyond the lungs and shows a complex interplay with the human host that is not always correlated with its entry receptor (ACE2) expression levels. Many studies identified viral components (RNA, proteins) of SARS-CoV-2 in multiple organs (pharynx, trachea, lungs, blood, heart, vessels, intestines, brain, male genitals and kidneys) and body fluids (mucus, saliva, urine, cerebrospinal fluid, semen and breast milk). However, besides the lungs, researchers were only able to detect infectious virus in stool and urine in a limited set of SARS-CoV-2 patients. By combining these studies, our study provides an eagle's view on the current status of SARS-CoV-2 pathogenesis and lays the foundation for better diagnosis and treatment of COVID-19 patients. Since SARS-CoV-2 appeared in the human population, the scientific community has scrambled to gather as much information as possible to find good strategies for the containment and treatment of this pandemic virus. Here, we performed a systematic review of the current (pre)published SARS-CoV-2 literature with a focus on the evidence concerning SARS-CoV-2 distribution in human tissues and viral shedding in body fluids. In addition, this evidence is aligned with published ACE2 entry-receptor (single cell) expression data across the human body to construct a viral distribution and ACE2 receptor body map. We highlight the broad organotropism of SARS-CoV-2, as many studies identified viral components (RNA, proteins) in multiple organs, including the pharynx, trachea, lungs, blood, heart, vessels, intestines, brain, male genitals and kidneys. This also implicates the presence of viral components in various body fluids such as mucus, saliva, urine, cerebrospinal fluid, semen and breast milk. The main SARS-CoV-2 entry receptor, ACE2, is expressed at different levels in multiple tissues throughout the human body, but its expression levels do not always correspond with SARS-CoV-2 detection, indicating that there is a complex interplay between virus and host. Together, these data shed new light on the current view of SARS-CoV-2 pathogenesis and lay the foundation for better diagnosis and treatment of COVID-19 patients

Maternal age effects on myometrial expression of contractile proteins, uterine gene expression, and contractile activity during labor in the rat

Advanced maternal age of first time pregnant mothers is associated with prolonged and dysfunctional labor and significant risk of emergency cesarean section. We investigated the influence of maternal age on myometrial contractility, expression of contractile associated proteins (CAPs), and global gene expression in the parturient uterus. Female Wistar rats either 8 (YOUNG n = 10) or 24 (OLDER n = 10) weeks old were fed laboratory chow, mated, and killed during parturition. Myometrial strips were dissected to determine contractile activity, cholesterol (CHOL) and triglycerides (TAG) content, protein expression of connexin-43 (GJA1), prostaglandin endoperoxide synthase 2 (PTGS2), and caveolin 1 (CAV-1). Maternal plasma concentrations of prostaglandins PGE2, PGF2a, and progesterone were determined by RIA. Global gene expression in uterine samples was compared using Affymetrix Genechip Gene 2.0 ST arrays and Ingenuity Pathway analysis (IPA). Spontaneous contractility in myometrium exhibited by YOUNG rats was threefold greater than OLDER animals (P < 0.027) but maternal age had no significant effect on myometrial CAP expression, lipid profiles, or pregnancy related hormones. OLDER myometrium increased contractile activity in response to PGF2a, phenylephrine, and carbachol, a response absent in YOUNG rats (all P < 0.002). Microarray analysis identified that maternal age affected expression of genes related to immune and inflammatory responses, lipid transport and metabolism, steroid metabolism, tissue remodeling, and smooth muscle contraction. In conclusion YOUNG laboring rat myometrium seems primed to contract maximally, whereas activity is blunted in OLDER animals and requires stimulation to meet contractile potential. Further work investigating maternal age effects on myometrial function is required with focus on lipidmetabolism and inflammatory pathways

Hunting for Serine 276-Phosphorylated p65

The transcription factor nuclear factor kappaB (NF-κB) is one of the central mediators of inflammatory gene expression. Several posttranslational modifications of NF-κB, regulating its transactivation ability, have been described. Especially phosphorylation of the NF-κB subunit p65 has been investigated in depth and several commercial phosphospecific antibodies, targeting selected p65 residues, are available. One of the p65 residues, that is subject to phosphorylation by protein kinase A (PKA) as well as by mitogen-stimulated kinase-1 (MSK-1), is the serine at position 276. Here, we have performed a detailed analysis of the performance of the most commonly used commercial anti-P-p65 Ser276 antibodies. Our findings indicate that at least three widely used anti-P-p65 Ser276 antibodies do not detect p65 in vivo via Western Blot, but instead crossreact with PKA-regulated proteins. As PKA is one of the main kinases responsible for phosphorylation of p65 at Ser276, this observation warrants cautious interpretation of data generated using the tested antibodies