SARS-CoV-2 variant Alpha has a spike-dependent replication advantage over the ancestral B.1 strain in human cells with low ACE2 expression

Epidemiological data demonstrate that Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) Alpha and Delta are more transmissible, infectious, and pathogenic than previous variants. Phenotypic properties of VOC remain understudied. Here, we provide an extensive functional study of VOC Alpha replication and cell entry phenotypes assisted by reverse genetics, mutational mapping of spike in lentiviral pseudotypes, viral and cellular gene expression studies, and infectivity stability assays in an enhanced range of cell and epithelial culture models. In almost all models, VOC Alpha spread less or equally efficiently as ancestral (B.1) SARS-CoV-2. B.1. and VOC Alpha shared similar susceptibility to serum neutralization. Despite increased relative abundance of specific sgRNAs in the context of VOC Alpha infection, immune gene expression in infected cells did not differ between VOC Alpha and B.1. However, inferior spreading and entry efficiencies of VOC Alpha corresponded to lower abundance of proteolytically cleaved spike products presumably linked to the T716I mutation. In addition, we identified a bronchial cell line, NCI-H1299, which supported 24-fold increased growth of VOC Alpha and is to our knowledge the only cell line to recapitulate the fitness advantage of VOC Alpha compared to B.1. Interestingly, also VOC Delta showed a strong (595-fold) fitness advantage over B.1 in these cells. Comparative analysis of chimeric viruses expressing VOC Alpha spike in the backbone of B.1, and vice versa, showed that the specific replication phenotype of VOC Alpha in NCI-H1299 cells is largely determined by its spike protein. Despite undetectable ACE2 protein expression in NCI-H1299 cells, CRISPR/Cas9 knock-out and antibody-mediated blocking experiments revealed that multicycle spread of B.1 and VOC Alpha required ACE2 expression. Interestingly, entry of VOC Alpha, as opposed to B.1 virions, was largely unaffected by treatment with exogenous trypsin or saliva prior to infection, suggesting enhanced resistance of VOC Alpha spike to premature proteolytic cleavage in the extracellular environment of the human respiratory tract. This property may result in delayed degradation of VOC Alpha particle infectivity in conditions typical of mucosal fluids of the upper respiratory tract that may be recapitulated in NCI-H1299 cells closer than in highly ACE2-expressing cell lines and models. Our study highlights the importance of cell model evaluation and comparison for in-depth characterization of virus variant-specific phenotypes and uncovers a fine-tuned interrelationship between VOC Alpha- and host cell-specific determinants that may underlie the increased and prolonged virus shedding detected in patients infected with VOC Alpha

Beitraege zu Ergebnissen demografischer Forschung und zur Herausbildung und Entwicklung des Leitungs- und Planungssystems der DDR

SIGLEIAB-32121-DD AT 902 / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekDEGerman

Oxytocin and positive couple interaction affect the perception of wound pain in everyday life

A large body of animal and human laboratory research has linked social interaction and support to pain perception, with a possible role for the neuropeptide oxytocin as a neuroendocrine mediator. However so far, it has been unclear whether these effects translate to ecologically valid everyday life behavior and pain perception. In a randomized placebo-controlled study, a standard suction blister skin wound was induced to N = 80 romantic couples (N = 160 individuals). Couples then received intranasal oxytocin or placebo twice daily and were either instructed to perform a positive social interaction (partner appraisal task, PAT) once in the laboratory and two times during the following five days, or not. During these days, all participants reported their subjective pain levels multiple times a day using ecologically momentary assessment. Results from hierarchical linear modeling suggest that pain levels within the couples were inter-related. In men, but not in women, oxytocin reduced pain levels. Women reported lower pain levels in the group of positive social interaction, while this effect did not show in men. These results suggest that intranasal oxytocin might have sex-specific effects with pain reducing effects in men but the opposite effects in women. In contrast, especially women benefit from positive interaction in terms of dampened pain levels after positive interaction. The results add to the evidence for health-beneficial effects of positive couple interaction and point to underlying neuroendocrine mechanisms in everyday life pain specifically. The sex-specific effects, in particular, may have implications for psychopharmacological treatment of pain in men and women

Mobile application-based interventions for chronic pain patients: a systematic review and meta-analysis of effectiveness

Chronic pain is one of the major causes of disability in the general population. Even though there are effective treatment options available for reducing symptoms, these treatments often do not have consistent lasting effects. As the usage of mobile devices has increased enormously during the last few years, mobile application-based treatment options are widespread. Such app-based programs are not yet empirically proven but might enable patients to become more independent in their pain management in order to prevent relapse. The aim of this meta-analysis was to summarize the literature on mobile application-based interventions for chronic pain patients. Therefore, three electronic bibliographic databases, PubMed, PsycINFO, and Web of Science, were searched for studies that investigated the effectiveness of mobile application-based intervention for chronic pain on pain intensity. The final sample comprised twenty-two studies, with a total of 4679 individuals. Twelve of these twenty-two studies used a randomized control trial (RCT) design, while ten studies only used an observational design. For all twenty-two studies, a small but significant effect (d = −0.40) was found when compared to baseline measures or control groups. The results suggest that apps-based treatment can be helpful in reducing pain, especially in the long-term

Rapid Changes of mRNA-binding Protein Levels following Glucose and 3-Isobutyl-1-methylxanthine Stimulation of Insulinoma INS-1 Cells *S⃞

Glucose and cAMP-inducing agents such as 3-isobutyl-1-methylxanthine (IBMX) rapidly change the expression profile of insulin-producing pancreatic β-cells mostly through post-transcriptional mechanisms. A thorough analysis of these changes, however, has not yet been performed. By combining two-dimensional differential gel electrophoresis and mass spectrometry, we identified 165 spots, corresponding to 78 proteins, whose levels significantly change after stimulation of the β-cell model INS-1 cells with 25 mm glucose + 1 mm IBMX for 2 h. Changes in the expression of selected proteins were verified by one- and two-dimensional immunoblotting. Most of the identified proteins are novel targets of rapid regulation in β-cells. The transcription inhibitor actinomycin D failed to block changes in two-thirds of the spots, supporting their post-transcriptional regulation. More spots changed in response to IBMX than to glucose alone conceivably because of phosphorylation. Fourteen mRNA- binding proteins responded to stimulation, thus representing the most prominent class of rapidly regulated proteins. Bioinformatics analysis indicated that the mRNA 5′- and 3′-untranslated regions of 22 regulated proteins contain potential binding sites for polypyrimidine tract-binding protein 1, which promotes mRNA stability and translation in stimulated β-cells. Overall our findings support the idea that mRNA-binding proteins play a major role in rapid adaptive changes in insulin-producing cells following their stimulation with glucose and cAMP-elevating agents

Functional comparison of MERS-coronavirus lineages reveals increased replicative fitness of the recombinant lineage 5.

Middle East respiratory syndrome coronavirus (MERS-CoV) is enzootic in dromedary camels across the Middle East and Africa. Virus-induced pneumonia in humans results from animal contact, with a potential for limited onward transmission. Phenotypic changes have been suspected after a novel recombinant clade (lineage 5) caused large nosocomial outbreaks in Saudi Arabia and South Korea in 2016. However, there has been no functional assessment. Here we perform a comprehensive in vitro and ex vivo comparison of viruses from parental and recombinant virus lineages (lineage 3, n = 7; lineage 4, n = 8; lineage 5, n = 9 viruses) from Saudi Arabia, isolated immediately before and after the shift toward lineage 5. Replication of lineage 5 viruses is significantly increased. Transcriptional profiling finds reduced induction of immune genes IFNB1, CCL5, and IFNL1 in lung cells infected with lineage 5 strains. Phenotypic differences may be determined by IFN antagonism based on experiments using IFN receptor knock out and signaling inhibition. Additionally, lineage 5 is more resilient against IFN pre-treatment of Calu-3 cells (ca. 10-fold difference in replication). This phenotypic change associated with lineage 5 has remained undiscovered by viral sequence surveillance, but may be a relevant indicator of pandemic potential

Evaluation of Potential PET Imaging Probes for the Orexin 2 Receptors

A wide range of central nervous system (CNS) disorders, particularly those related to sleep, are associated with the abnormal function of orexin (OX) receptors. Several orexin receptor antagonists have been reported in recent years, but currently there are no imaging tools to probe the density and function of orexin receptors in vivo. To date there are no published data on the pharmacokinetics (PK) and accumulation of some lead orexin receptor antagonists. Evaluation of CNS pharmacokinetics in the pursuit of positron emission tomography (PET) radiotracer development could be used to elucidate the association of orexin receptors with diseases and to facilitate the drug discovery and development. To this end, we designed and evaluated carbon-11 labeled compounds based on diazepane orexin receptor antagonists previously described. One of the synthesized compounds, [11C]CW4, showed high brain uptake in rats and further evaluated in non-human primate (NHP) using PET-MR imaging. PET scans performed in a baboon showed appropriate early brain uptake for consideration as a radiotracer. However, [11C]CW4 exhibited fast kinetics and high nonspecific binding, as determined after co-administration of [11C]CW4 and unlabeled CW4. These properties indicate that [11C]CW4 has excellent brain penetrance and could be used as a lead compound for developing new CNS-penetrant PET imaging probes of orexin receptors.Chemistry and Chemical Biolog

Oxytocin and positive couple interaction affect the perception of wound pain in everyday life

A large body of animal and human laboratory research has linked social interaction and support to pain perception, with a possible role for the neuropeptide oxytocin as a neuroendocrine mediator. However so far, it has been unclear whether these effects translate to ecologically valid everyday life behavior and pain perception. In a randomized placebo-controlled study, a standard suction blister skin wound was induced to N = 80 romantic couples (N = 160 individuals). Couples then received intranasal oxytocin or placebo twice daily and were either instructed to perform a positive social interaction (partner appraisal task, PAT) once in the laboratory and two times during the following five days, or not. During these days, all participants reported their subjective pain levels multiple times a day using ecologically momentary assessment. Results from hierarchical linear modeling suggest that pain levels within the couples were inter-related. In men, but not in women, oxytocin reduced pain levels. Women reported lower pain levels in the group of positive social interaction, while this effect did not show in men. These results suggest that intranasal oxytocin might have sex-specific effects with pain reducing effects in men but the opposite effects in women. In contrast, especially women benefit from positive interaction in terms of dampened pain levels after positive interaction. The results add to the evidence for health-beneficial effects of positive couple interaction and point to underlying neuroendocrine mechanisms in everyday life pain specifically. The sex-specific effects, in particular, may have implications for psychopharmacological treatment of pain in men and women

Frühmobilisation auf der Intensivstation : sind robotergestützte Systeme die Zukunft?

'''Hintergrund:''' Bei etwa 43% aller Überlebenden der Intensivmedizin wird ein erworbenes Syndrom an Muskelschwäche beobachtet, welches Überleben und Lebensqualität vermindert. Da kausale Therapieoptionen bisher fehlen, stehen die Vermeidung der bekannten Risikofaktoren und Frühmobilisation im Vordergrund. Robotische Unterstützungssysteme werden vermehrt in der Mobilisation erprobt. '''Ziel der Arbeit:''' In diesem Übersichtsartikel wird die aktuelle Evidenz von Frühmobilisation von kritisch Kranken zusammengefasst und der Stellenwert robotischer Assistenzsysteme für Mobilisation diskutiert. '''Ergebnisse:''' Mobilisation sollte auf der Intensivstation nachMöglichkeit früh begonnen werden. Hierunter wird der Beginn in den ersten 72 h nach der Aufnahme auf die Intensivstation verstanden. Physiotherapeutische Interventionen während des Intensivaufenthalts zeigen positive Effekte auf die Lebensqualität von PatientInnen, auf die Dauer von invasiver Beatmung, Intensivaufenthalt und Delir. Strukturierte Behandlungsprotokolle führen zu mehr aktiver Mobilisation, höherer Mobilität und häufigerer funktioneller Unabhängigkeit bei Entlassung aus dem Krankenhaus. Nach Schlaganfällen erhöhen zusätzliche robotergestützte Therapieeinheiten insbesondere bei stärker eingeschränkten PatientInnen die Rate an Rückkehrern zum selbstständigen Gehen, scheinen sicher und verbesserten in kleinen Studien Muskelkraft und Lebensqualität. '''Schlussfolgerung:''' Frühmobilisation verbessert das Outcome von kritisch Erkrankten. Robotische Systeme unterstützen das Gangtraining nach einem Schlaganfall und werden auf der Intensivstation in ersten Studien zu Vertikalisierung und Frühmobilisation untersucht