Unmasking features of the auto-epitope essential for β(1)-adrenoceptor activation by autoantibodies in chronic heart failure

AIMS: Chronic heart failure (CHF) can be caused by autoantibodies stimulating the heart via binding to first and/or second extracellular loops of cardiac β(1)-adrenoceptors. Allosteric receptor activation depends on conformational features of the autoantibody binding site. Elucidating these features will pave the way for the development of specific diagnostics and therapeutics. Our aim was (i) to fine-map the conformational epitope within the second extracellular loop of the human β(1)-adrenoceptor (β(1) EC(II)) that is targeted by stimulating β(1)-receptor (auto)antibodies and (ii) to generate competitive cyclopeptide inhibitors of allosteric receptor activation, which faithfully conserve the conformational auto-epitope. METHODS AND RESULTS: Non-conserved amino acids within the β(1) ECII loop (compared with the amino acids constituting the ECII loop of the β(2)-adrenoceptor) were one by one replaced with alanine; potential intra-loop disulfide bridges were probed by cysteine-serine exchanges. Effects on antibody binding and allosteric receptor activation were assessed (i) by (auto)antibody neutralization using cyclopeptides mimicking β(1) ECII ± the above replacements, and (ii) by (auto)antibody stimulation of human β(1)-adrenoceptors bearing corresponding point mutations. With the use of stimulating β(1)-receptor (auto)antibodies raised in mice, rats, or rabbits and isolated from exemplary dilated cardiomyopathy patients, our series of experiments unmasked two features of the β(1) ECII loop essential for (auto)antibody binding and allosteric receptor activation: (i) the NDPK(211-214) motif and (ii) the intra-loop disulfide bond C(209)↔C(215). Of note, aberrant intra-loop disulfide bond C(209)↔C(216) almost fully disrupted the functional auto-epitope in cyclopeptides. CONCLUSIONS: The conformational auto-epitope targeted by cardio-pathogenic β(1)-receptor autoantibodies is faithfully conserved in cyclopeptide homologues of the β(1) EC(II) loop bearing the NDPK(211-214) motif and the C(209)↔C(215) bridge while lacking cysteine C(216). Such molecules provide promising tools for novel diagnostic and therapeutic approaches in β(1)-autoantibody-positive CHF

Chronic Cellular Imaging of Entire Cortical Columns in Awake Mice Using Microprisms

SummaryTwo-photon imaging of cortical neurons in vivo has provided unique insights into the structure, function, and plasticity of cortical networks, but this method does not currently allow simultaneous imaging of neurons in the superficial and deepest cortical layers. Here, we describe a simple modification that enables simultaneous, long-term imaging of all cortical layers. Using a chronically implanted glass microprism in barrel cortex, we could image the same fluorescently labeled deep-layer pyramidal neurons across their entire somatodendritic axis for several months. We could also image visually evoked and endogenous calcium activity in hundreds of cell bodies or long-range axon terminals, across all six layers in visual cortex of awake mice. Electrophysiology and calcium imaging of evoked and endogenous activity near the prism face were consistent across days and comparable with previous observations. These experiments extend the reach of in vivo two-photon imaging to chronic, simultaneous monitoring of entire cortical columns.Video Abstrac

When separation is not the answer : breastfeeding mothers and infants affected by COVID-19

The World Health Organisation (WHO) has provided detailed guidance on the care of infants of women who are a person under investigation (PUI) or confirmed to have COVID-19, which supports immediate postpartum mother-infant contact and breastfeeding with appropriate respiratory precautions. Although many countries have followed WHO guidance, others have implemented infection prevention and control policies (IPC) that impose varying levels of postpartum separation and discourage or prohibit breastfeeding or provision of expressed breastmilk. These policies aim to protect infants from the potential harm of infection from their mothers, yet they may fail to fully account for the impact of separation. Global COVID-19 data are suggestive of potentially lower susceptibility and a typically milder course of disease among children, although the potential for severe disease in infancy remains. Separation causes cumulative harms, including disrupting breastfeeding and limiting its protection against infectious disease, which has disproportionate impacts on vulnerable infants. Separation also presumes the replaceability of breastfeeding – a risk that is magnified in emergencies. Moreover, separation does not ensure lower viral exposure during hospitalizations and post-discharge, and contributes to the burden on overwhelmed health systems. Finally, separation magnifies maternal health consequences of insufficient breastfeeding and compounds trauma in communities who have experienced long-standing inequities and violence, including family separation. Taken together, separating PUI/confirmed SARS-CoV-2 positive mothers and their infants may lead to excess preventable illnesses and deaths among infants and women around the world. Health services must consider the short-and-long-term impacts of separating mothers and infants in their policies. This article is protected by copyright

Electron dynamics in planar radio frequency magnetron plasmas: II. Heating and energization mechanisms studied via a 2d3v particle-in-cell/Monte Carlo code

The present work investigates electron transport and heating mechanisms using an (r, z) particle-in-cell (PIC) simulation of a typical rf-driven axisymmetric magnetron discharge with a conducting target. It is shown that for the considered magnetic field topology the electron current flows through different channels in the (r, z) plane: a ``transverse'' one, which involves current flow through the electrons' magnetic confinement region (EMCR) above the racetrack, and two ''longitudinal'' ones. Electrons gain energy from the electric field along these channels following various mechanisms, which are rather distinct from those sustaining dc-powered magnetrons. The longitudinal power absorption involves mirror-effect heating (MEH), nonlinear electron resonance heating (NERH), magnetized bounce heating (MBH), and the heating by the ambipolar field at the sheath-presheath interface. The MEH and MBH represent two new mechanisms missing from the previous literature. The MEH is caused by a reversed electric field needed to overcome the mirror force generated in a nonuniform magnetic field to ensure sufficient flux of electrons to the powered electrode, and the MBH is related to a possibility for an electron to undergo multiple reflections from the expanding sheath in the longitudinal channels connected by the arc-like magnetic field. The electron heating in the transverse channel is caused mostly by the essentially collisionless Hall heating in the EMCR above the racetrack, generating a strong ExB azimuthal drift velocity. The latter mechanism results in an efficient electron energization, i.e., energy transfer from the electric field to electrons in the inelastic range. Since the main electron population energized by this mechanism remains confined within the discharge for a long time, its contribution to the ionization processes is dominant

Nagetier-übertragene Zoonosen: Beispiele aus Untersuchungen in Süd- und Westdeutschland

Zusammenfassung Nagetiere und andere Kleinsäuger können eine Vielzahl von Krankheitserregern, RNA- und DNA-Viren, Bakterien und Parasiten, auf den Menschen übertragen, die teilweise lebensbedrohliche Erkrankungen hervorrufen. In der folgenden Übersicht soll erstmals ein Überblick über Ergebnisse aus drei Untersuchungen in Deutschland gegeben werden: eine Studie in drei Landkreisen Bayerns von 2001-2004, Untersuchungen in einem Freilandgehege im Rahmen eines Tularämieausbruchs in Niedersachsen im Jahr 2005, und schließlich im Jahr 2007 eine Untersuchung an einem Truppenübungsplatz in Baden-Württemberg. Es wurde dabei exemplarisch die Verbreitung von Zoonoseerregern in Nagetieren und anderen Kleinsäugern näher untersucht, von drei Viren (Hantaviren, Kuhpockenvirus, Frühsommer- Meningo-Enzephalitis-Virus) und vier Bakterien (Leptospiren, Francisellen, Borrelien und Rickettsien). Die hier zusammengefassten Erkenntnisse sind ein erster wichtiger Schritt auf dem Weg zur Erstellung von Verbreitungskarten für die genannten humanpathogenen Zoonoseerreger in ihren Reservoirwirten und der Definition von entsprechenden Risikogebieten. Diese Arbeit soll zudem einen Beitrag leisten, einen Anstoß zu verstärkter Zusammenarbeit von Zoologen, Ökologen, Virologen, Human- und Veterinärmedizinern, Mikrobiologen, Parasitologen, Genetikern, Epidemiologen, Forstwissenschaftlern und Klimaforschern zu geben

Tick species from cattle in the Adama Region of Ethiopia and pathogens detected

Ticks will diminish productivity among farm animals and transmit zoonotic diseases. We conducted a study to identify tick species infesting slaughter bulls from Adama City and to screen them for tick-borne pathogens. In 2016, 291 ticks were collected from 37 bulls in Adama, which were ready for slaughter. Ticks were identified morphologically. Total genomic DNA was extracted from ticks and used to test for Rickettsia spp. with real-time PCR. Species identification was done by phylogenetic analysis using sequencing that targeted the 23S-5S intergenic spacer region and ompA genes. Four tick species from two genera, Amblyomma and Rhipicephalus, were identified. Amblyomma cohaerens was the dominant species (n = 241, 82.8%), followed by Amblyomma variegatum (n = 22, 7.5%), Rhipicephalus pulchellus (n = 19, 6.5%), and Rhipicephalus decoloratus (n = 9, 3.0%). Among all ticks, 32 (11%) were positive for Rickettsia spp. and 15 (5.2%) of these were identified as R. africae comprising at least two genetic clades, occurring in A. variegatum (n = 10) and A. cohaerens (n = 5). The remainder of Rickettsia-positive samples could not be amplified due to low DNA yield. Furthermore, another 15 (5.2%) samples carried other pathogenic bacteria: Ehrlichia ruminantium (n = 9; 3.1%) in A. cohaerens, Ehrlichia sp. (n = 3; 1%) in Rh. pulchellus and A. cohaerens, Anaplasma sp. (n = 1; 0.5%) in A. cohaerens, and Neoehrlichia mikurensis (n = 2; 0.7%) in A. cohaerens. All ticks were negative for Bartonella spp., Babesia spp., Theileria spp., and Hepatozoon spp. We reported for the first time E. ruminatium, N. mikurensis, Ehrlichia sp., and Anaplasma sp. in A. cohaerens. Medically and veterinarily important pathogens were mostly detected from A. variegatum and A. cohaerens. These data are relevant for a One-health approach for monitoring and prevention of tick-borne disease transmission

Effect of radiotherapy on local recurrence, distant metastasis and overall survival in 1200 extremity soft tissue sarcoma patients.:Retrospective analysis using IPTW-adjusted models

Background and purpose: Neoadjuvant (NRTX) and adjuvant radiotherapy (ARTX) reduce local recurrence (LR) risk in extremity soft tissue sarcoma (eSTS), yet their impact on distant metastasis (DM) and overall survival (OS) is less well defined. This study aimed at analysing the influence of NRTX/ARTX on all three endpoints using a retrospective, multicentre eSTS cohort. Materials and methods: 1200 patients (mean age: 60.7 ± 16.8 years; 44.4 % females) were retrospectively included, treated with limb sparing surgery and curative intent for localised, high grade (G2/3) eSTS. 194 (16.2 %), 790 (65.8 %), and 216 (18.0 %) patients had received NRTX, ARTX and no RTX, respectively. For the resulting three groups (no RTX vs. NRTX, no RTX vs. ARTX, NRTX vs. ARTX) Fine&amp;Gray models for LR and DM, and Cox-regression models for OS were calculated, with IPTW-modelling adjusting for imbalances between groups. Results: In the IPTW-adjusted analysis, NRTX was associated with lower LR-risk in comparison to no RTX (SHR [subhazard ratio]: 0.236; p = 0.003), whilst no impact on DM-risk (p = 0.576) or OS (p = 1.000) was found. IPTW-weighted analysis for no RTX vs. ARTX revealed a significant positive association between ARTX and lower LR-risk (SHR: 0.479, p = 0.003), but again no impact on DM-risk (p = 0.363) or OS (p = 0.534). IPTW-weighted model for NRTX vs. ARTX showed significantly lower LR-risk for NRTX (SHR for ARTX: 3.433; p = 0.003) but no difference regarding DM-risk (p = 1.000) or OS (p = 0.639). Conclusion: NRTX and ARTX are associated with lower LR-risk, but do not seem to affect DM-risk or OS. NRTX may be favoured over ARTX as our results indicate better local control rates.</p

The 'lost child' as figure of trauma and recovery in early post-war cinema: Fred Zinnemann's The Search (1948) and Natan Gross's Unzere Kinder (1948)

The article examines the figure of the ‘lost child’ in feature films of the immediate post-war period. The figure’s enormous symbolic value as innocent victim and future generation, granted the ‘lost child’ a key position in post-war discourse, including films which tried to grapple with the moral and physical destruction of the continent after 1945. National film industries, particularly of the perpetrator nation, employed the ‘lost child’ for genre stories in which the post-war chaos is being mastered and a new, masculine national self is re-built. However, films made by victim groups outside a national context rely on the ‘lost child’ to broach the destruction of their identity by war and persecution. Analysing two films, Fred Zinnemann’s The Search (1948) and Nata Gross’s Unzere Kinder (1948), I argue that they use the child figure to deal with traumatization and make it part of the reconstruction of communal intergenerational relations. This does not result in stories of masculine mastery but in narratives that incorporate moments of trauma process emerging around destroyed mother-child relations. The films, encoding traumatization in film language, develop a rich cinematic language along questions of identity and form a first instance of posttraumatic cinema

Re-Emergence of Crimean-Congo Hemorrhagic Fever Virus in Central Africa

Crimean-Congo hemorrhagic fever virus (CCHFV) is transmitted to humans through tick-bite or contact with infected blood or tissues from livestock, the main vertebrate hosts in a peri-domestic natural cycle. With numerous outbreaks, a high case fatality rate (3%–30%) and a high risk for nosocomial transmission, CCHFV became a public health concern in Europe and Asia. However virus surveillance in Africa is difficult due to the limited sanitary facilities. Especially, CCHFV occurrence in Central Africa is very poorly described and seems highly in contrast with the temperate to dry environments to which the virus is usually associated with. We described a single human infection that occurred in Democratic Republic of the Congo after nearly 50 years of absence. The phylogenetic analysis suggests that CCHFV enzootic circulation in the area is still ongoing despite the absence of notification, and thus reinforces the need for the medical workers and authorities to be aware of the outbreak risk. The source of infection seemed associated with a forest environment while no link with the usual agro-pastoral risk factors could be identified. More accurate ecological data about CCHFV enzootic cycle are required to assess the risk of emergence in developing countries subjected to deforestation

Measurement of the Neutron Spin Structure Function g1ng_1^n with a Polarized ^3He Target

Results are reported from the HERMES experiment at HERA on a measurement of the neutron spin structure function $g_1^n(x,Q^2)$ in deep inelastic scattering using 27.5 GeV longitudinally polarized positrons incident on a polarized $^3$He internal gas target. The data cover the kinematic range $0.023<x<0.6$ and $1 (GeV/c)^2 < Q^2 <15 (GeV/c)^2$. The integral $\int_{0.023}^{0.6} g_1^n(x) dx$ evaluated at a fixed $Q^2$ of $2.5 (GeV/c)^2$ is $-0.034\pm 0.013(stat.)\pm 0.005(syst.)$. Assuming Regge behavior at low $x$, the first moment $\Gamma_1^n=\int_0^1 g_1^n(x) dx$ is $-0.037\pm 0.013(stat.)\pm 0.005(syst.)\pm 0.006(extrapol.)$.Comment: 4 pages TEX, text available at http://www.krl.caltech.edu/preprints/OAP.htm