Action potentials in primary osteoblasts and in the MG-63 osteoblast-like cell line

Whole-cell patch-clamp analysis revealed a resting membrane potential of −60 mV in primary osteoblasts and in the MG-63 osteoblast-like cells. Depolarization-induced action potentials were characterized by duration of 60 ms, a minimal peak-to-peak distance of 180 ms, a threshold value of −20 mV and a repolarization between the spikes to −45 mV. Expressed channels were characterized by application of voltage pulses between −150 mV and 90 mV in 10 mV steps, from a holding potential of −40 mV. Voltages below −60 mV induced an inward current. Depolarizing voltages above −30 mV evoked two currents: (a) a fast activated and inactivated inward current at voltages between −30 and 30 mV, and (b) a delayed-activated outward current that was induced by voltages above −30 mV. Electrophysiological and pharmacological parameters indicated that hyperpolarization activated strongly rectifying K+ (Kir) channels, whereas depolarization activated tetrodotoxin sensitive voltage gated Na+ (Nav) channels as well as delayed, slowly activated, non-inactivating, and tetraethylammonium sensitive voltage gated K+ (Kv) channels. In addition, RT-PCR showed expression of Nav1.3, Nav1.4, Nav1.5, Nav1.6, Nav1.7, and Kir2.1, Kir2.3, and Kir2.4 as well as Kv2.1. We conclude that osteoblasts express channels that allow firing of action potentials. The final publication is available at Springer via https://doi.org/10.1007/s10863-011-9354-7.BMBF/NANOTOMEDFG/Transregio37Boehringer Ingelheim International GmbHBMBF/NANOTOMEDFG/Transregio/37Boehringer Ingelheim International Gmb

Spielen, Bewegen, Erkunden - Praktiken der Raumaneignung von Grundschulkindern in der Draußenschule

"In den Sozial- und Erziehungswissenschaften ist seit den 1990er-Jahren zunehmend eine neue Aufmerksamkeit für den Raum (spatial turn) zu beobachten, die sich auch auf die Ordnung kindlicher Lern- und Lebensräume richtet. Einen prominenten Stellenwert nimmt insbesondere die Betrachtung des Schulraumes ein. Mit dem Diskurs um die sozialräumliche Öffnung von Schule geraten dabei auch außerschulische Lernorte in den Fokus. Weithin vernachlässigt werden bislang jedoch zwei Perspektiven: 1) Die Sicht der Kinder auf den Unterricht an außerschulischen Lernorten und 2) die Analyse kindlicher Praktiken der Aneignung des (Draußen-)Schulraumes. Dem widmet sich der vorliegende Beitrag, indem er, ausgehend von raum- und aneignungstheoretischen Ansätzen sowie auf der empirischen Basis von Gruppengesprächen mit Grundschulkindern, deren Raumaneignungspraktiken in der Draußenschule - dem regelmäßigen Unterricht an außerschulischen Lernorten - diskutiert." (Autorenreferat)"The observable new attention for space in social and educational sciences since the 1990s (spatial turn) focuses more and more on the spatial constitution of children's learning and living spaces. Of relative importance is especially the classroom as a learning environment. But the discourse about the sociospatial opening of school also brings extracurricular educational facilities into focus. So far, two perspectives are still widely neglected: 1) The children’s point of view concerning lessons at extracurricular learning places, and 2) the analysis of the children’s practices of appropriation of the (outdoor) school space. The present article uses approaches of spatial and activity theory as well as group discussions with primary school children as empirical basis. It is about to shed light on this topic by discussing the children's practices of appropriating space in the German Outdoor School (Draußenschule) - the regular instruction at extracurricular learning places." (author's abstract

Vorstellung des Studienreformforums: Bisherige Arbeit und aktuelle Beiträge

Das Studienreform-Forum befasst sich einerseits mit der Systematisierung von Studienreformen und Studienreform-Vorhaben, andererseits mit Grundsatzfragen der Studienreform. Beides zusammen bildet die Grundlage zur Weiterentwicklung von Studiengängen.Im Jahr 2020 hat das Studienreform-Forum erneut zur Einsendung von Beiträgen zu diesen Fragen aufgerufen. Angesichts der Pandemie wurde dieser Aufruf verlängert und parallel eine Initiative zur Dokumentation und Auswertung der Lehre unter Pandemiebedingungen gestartet, deren Ergebnisse mittelfristig mit den übrigen Beiträgen in Bezug gesetzt werden sollen.Dieser Artikel dokumentiert die auf den Call for Papers eingesandten Beiträge

IND-Enabling Studies for a Clinical Trial to Genetically Program a Persistent Cancer-Targeted Immune System

PURPOSE: To improve persistence of adoptively transferred T-cell receptor (TCR)-engineered T cells and durable clinical responses, we designed a clinical trial to transplant genetically-modified hematopoietic stem cells (HSCs) together with adoptive cell transfer of T cells both engineered to express an NY-ESO-1 TCR. Here, we report the preclinical studies performed to enable an investigational new drug (IND) application. EXPERIMENTAL DESIGN: HSCs transduced with a lentiviral vector expressing NY-ESO-1 TCR and the PET reporter/suicide gene HSV1-sr39TK and T cells transduced with a retroviral vector expressing NY-ESO-1 TCR were coadministered to myelodepleted HLA-A2/Kb mice within a formal Good Laboratory Practice (GLP)-compliant study to demonstrate safety, persistence, and HSC differentiation into all blood lineages. Non-GLP experiments included assessment of transgene immunogenicity and in vitro viral insertion safety studies. Furthermore, Good Manufacturing Practice (GMP)-compliant cell production qualification runs were performed to establish the manufacturing protocols for clinical use. RESULTS: TCR genetically modified and ex vivo-cultured HSCs differentiated into all blood subsets in vivo after HSC transplantation, and coadministration of TCR-transduced T cells did not result in increased toxicity. The expression of NY-ESO-1 TCR and sr39TK transgenes did not have a detrimental effect on gene-modified HSC's differentiation to all blood cell lineages. There was no evidence of genotoxicity induced by the lentiviral vector. GMP batches of clinical-grade transgenic cells produced during qualification runs had adequate stability and functionality. CONCLUSIONS: Coadministration of HSCs and T cells expressing an NY-ESO-1 TCR is safe in preclinical models. The results presented in this article led to the FDA approval of IND 17471

Calcitonin gene-related peptide inhibits Langerhans cell-mediated HIV-1 transmission.

International audienceUpon its mucosal entry, human immunodeficiency virus type 1 (HIV-1) is internalized by Langerhans cells (LCs) in stratified epithelia and transferred locally to T cells. In such epithelia, LCs are in direct contact with peripheral neurons secreting calcitonin gene-related peptide (CGRP). Although CGRP has immunomodulatory effects on LC functions, its potential influence on the interactions between LCs and HIV-1 is unknown. We show that CGRP acts via its receptor expressed by LCs and interferes with multiple steps of LC-mediated HIV-1 transmission. CGRP increases langerin expression, decreases selected integrins, and activates NF-κB, resulting in decreased HIV-1 intracellular content, limited formation of LC-T cell conjugates, and elevated secretion of the CCR5-binding chemokine CCL3/MIP-1α. These mechanisms cooperate to efficiently inhibit HIV-1 transfer from LCs to T cells and T cell infection. In vivo, HIV-1 infection decreases CGRP plasma levels in both vaginally SHIV-challenged macaques and HIV-1-infected individuals. CGRP plasma levels return to baseline after highly active antiretroviral therapy. Our results reveal a novel path by which a peripheral neuropeptide acts at the molecular and cellular levels to limit mucosal HIV-1 transmission and suggest that CGRP receptor agonists might be used therapeutically against HIV-1

Workshop: Hochschuldidaktische Konsequenzen aus zwei Semestern Krisenlehre

Die Umstellung des Lehrbetriebs an den Hochschulen auf Online-Lehre hat nicht nur technische, sondern vor allem auch didaktische Herausforderungen mit sich gebracht, die vielfältig – und zum Teil sehr unterschiedlich – beantwortet wurden. An einer systematischen hochschulübergreifenden Auswertung fehlt es bislang aber noch. Im hir dokumentierten Workshop wurde der aktuelle Stand exemplarisch vorgestellt und diskutiert, wie es gelingen kann, dass die wertvollen Erfahrungen dieser Zeit nicht mit der Rückkehr zur Präsenzlehre verloren gehen

Autonomic modulation and antiarrhythmic therapy in a model of long QT syndrome type 3

AIMS: Clinical observations in patients with long QT syndrome carrying sodium channel mutations (LQT3) suggest that bradycardia caused by parasympathetic stimulation may provoke torsades de pointes (TdP). beta-Adrenoceptor blockers appear less effective in LQT3 than in other forms of the disease. METHODS AND RESULTS: We studied effects of autonomic modulation on arrhythmias in vivo and in vitro and quantified sympathetic innervation by autoradiography in heterozygous mice with a knock-in deletion (DeltaKPQ) in the Scn5a gene coding for the cardiac sodium channel and increased late sodium current (LQT3 mice). Cholinergic stimulation by carbachol provoked bigemini and TdP in freely roaming LQT3 mice. No arrhythmias were provoked by physical stress, mental stress, isoproterenol, or atropine. In isolated, beating hearts, carbachol did not prolong action potentials per se, but caused bradycardia and rate-dependent action potential prolongation. The muscarinic inhibitor AFDX116 prevented effects of carbachol on heart rate and arrhythmias. beta-Adrenoceptor stimulation suppressed arrhythmias, shortened rate-corrected action potential duration, increased rate, and minimized difference in late sodium current between genotypes. beta-Adrenoceptor density was reduced in LQT3 hearts. Acute beta-adrenoceptor blockade by esmolol, propranolol or chronic propranolol in vivo did not suppress arrhythmias. Chronic flecainide pre-treatment prevented arrhythmias (all P < 0.05). CONCLUSION: Cholinergic stimulation provokes arrhythmias in this model of LQT3 by triggering bradycardia. beta-Adrenoceptor density is reduced, and beta-adrenoceptor blockade does not prevent arrhythmias. Sodium channel blockade and beta-adrenoceptor stimulation suppress arrhythmias by shortening repolarization and minimizing difference in late sodium current.status: publishe

Increased Prevalence of Metabolic Syndrome in Patients with Acne Inversa

BACKGROUND: Acne inversa (AI; also designated as Hidradenitis suppurativa) is a common chronic inflammatory skin disease, localized in the axillary, inguinal and perianal skin areas that causes painful, fistulating sinuses with malodorous purulence and scars. Several chronic inflammatory diseases are associated with the metabolic syndrome and its consequences including arteriosclerosis, coronary heart disease, myocardial infraction, and stroke. So far, the association of AI with systemic metabolic alterations is largely unexplored. METHODS AND FINDINGS: A hospital-based case-control study in 80 AI patients and 100 age- and sex-matched control participants was carried out. The prevalence of central obesity (odds ratio 5.88), hypertriglyceridemia (odds ratio 2.24), hypo-HDL-cholesterolemia (odds ratio 4.56), and hyperglycemia (odds ratio 4.09) in AI patients was significantly higher than in controls. Furthermore, the metabolic syndrome, previously defined as the presence of at least three of the five alterations listed above, was more common in those patients compared to controls (40.0% versus 13.0%; odds ratio 4.46, 95% confidence interval 2.02 to 9.96; P<0.001). AI patients with metabolic syndrome also had more pronounced metabolic alterations than controls with metabolic syndrome. Interestingly, there was no correlation between the severity or duration of the disease and the levels of respective parameters or the number of criteria defining the metabolic syndrome. Rather, the metabolic syndrome was observed in a disproportionately high percentage of young AI patients. CONCLUSIONS: This study shows for the first time that AI patients have a high prevalence of the metabolic syndrome and all of its criteria. It further suggests that the inflammation present in AI patients does not have a major impact on the development of metabolic alterations. Instead, evidence is given for a role of metabolic alterations in the development of AI. We recommend monitoring of AI patients in order to correct their modifiable cardiovascular risk factors

Mosquitoes LTR Retrotransposons: A Deeper View into the Genomic Sequence of Culex quinquefasciatus

A set of 67 novel LTR-retrotransposon has been identified by in silico analyses of the Culex quinquefasciatus genome using the LTR_STRUC program. The phylogenetic analysis shows that 29 novel and putatively functional LTR-retrotransposons detected belong to the Ty3/gypsy group. Our results demonstrate that, by considering only families containing potentially autonomous LTR-retrotransposons, they account for about 1% of the genome of C. quinquefasciatus. In previous studies it has been estimated that 29% of the genome of C. quinquefasciatus is occupied by mobile genetic elements

The Digital MIQE Guidelines Update: Minimum Information for Publication of Quantitative Digital PCR Experiments for 2020

Digital PCR (dPCR) has developed considerably since the publication of the Minimum Information for Publication of Digital PCR Experiments (dMIQE) guidelines in 2013, with advances in instrumentation, software, applications, and our understanding of its technological potential. Yet these developments also have associated challenges; data analysis steps, including threshold setting, can be difficult and preanalytical steps required to purify, concentrate, and modify nucleic acids can lead to measurement error. To assist independent corroboration of conclusions, comprehensive disclosure of all relevant experimental details is required. To support the community and reflect the growing use of dPCR, we present an update to dMIQE, dMIQE2020, including a simplified dMIQE table format to assist researchers in providing key experimental information and understanding of the associated experimental process. Adoption of dMIQE2020 by the scientific community will assist in standardizing experimental protocols, maximize efficient utilization of resources, and further enhance the impact of this powerful technology