Entwicklung einer neuen Echtzeit-Methode zur Untersuchung des erytrozytärer Lebenszyklus von Plasmodium falciparum mit Schwingquarzbiosensoren

Previous studies carried out by the Biosensor Research Group of the Institute of Clinical and Experimental Transfusion Medicine at the Tübingen University had successfully demonstrated determination of various blood types through the interaction of erythrocytes with immobilised antibodies on a QCM. These studies have opened the possibility to test new applications of this technique for the study of infectious diseases. In our case, studies of the erythrocyte life cycle of P. falciparum, particularly during the last six hours preceding the merozoite release and studies related to reinfection of erythrocytes by merozoites were the focus of our investigation. Release of malaria parasites of P. falciparum from infected erythrocyte at the end of their asexual erythrocyte cycle occurs approximately every 48 hours, asynchronously in parasite culture and synchronously in vivo. This process is poorly studied due to: (I) merozoite release is a very short event (<60 s), (II) schizont stages have high sensitivity for culture conditions like pH, medium, osmotic pressure, gas atmosphere and temperature. (III) Schizont conditions are not easy to maintain under the miscrocope, making miscroscopical real time observation of the release difficult. The aim of the presented project was to study the release of Plasmodium falciparum merozoites from erythrocytes with the QCM sensor technique. In this way, the frequency shift due to the change of mass associated to the merozoite release and, on a second QCM, the reinfection of healthy erythrocytes are monitored in real time. Our QCM experiments included the following stages: (I) Adaptation and optimisation of the immobilisation of biological layers to capture the erythrocytes on the quartz. (II) Optimisation of parasite culture conditions in a QCM chamber. (III) Observation of the frequency signal both for infected and non-infected erythrocytes samples and correlation of the signal with the release of merozoites. (IV) Reinfection of healthy erythrocytes on a second quartz within one QCM system. (V) Test of inhibition of merozoite release and reinfection by antimalarial compounds. The results showed that there was significant increase of ~1000 Hz for QCM with infected erythrocyte compared to QCM with healthy erythrocytes, where the frequency remained stable. Microscopical observation of the quartz surface at different times during the experiment (TEM and optical) demonstrated a correlation between the increase in frequency and merozoite release. At this point, approximately more than 80% of the infected erythrocytes on the quartz are involved in the release. Reinfection of new erythrocytes was observed on a second QCM. pH of the system (7.2), Temperature (37°C+/-0.1), flow of the medium (9 µL/min), sterility of the process (BactAlert), gas atmosphere (O2 5%, residual N2) were established to ensure parasite development and survival. External controls using flow cytometry 24 hours after the reinfection showed a parasitemia percentage of >1% in the erythrocytes infected in situ. Our results show, that the QCM technique is an appropriate and important new tool to elucidate the biology of the re-invasion process of Plasmodia.Frühere Studien, ausgeführt durch die Biosensor-Forschungsgruppe des Instituts für Klinische und Experimentelle Transfusionsmedizin an der Universität Tübingen, hatten die Blutgruppenbestimmung durch die Wechselwirkung von Erythrozyten mit immobilisierten Antikörpern auf einem Schwingquarzbiosensor (SQ) erfolgreich demonstriert. Diese Studien haben die Möglichkeit eröffnet, neue Anwendungen dieser Technik für die Untersuchung von Infektionskrankheiten zu evaluieren. In unserem Fall waren Studien des erythrozytären Lebenszyklus von P. falciparum, besonders die letzten sechs Stunden, die der Merozoiten-Freisetzung vorangehen, und Studien verbunden mit der Reinfektion durch Merozoiten der Schwerpunkt unserer Forschung. Die Freisetzung von Malaria-Parasiten von P. falciparum aus infizierten Erythrozyten am Ende ihres asexuellen erythrozytären Lebenzyklus erfolgt alle 48 Stunden und ist asynchron in Parasiten-Kultur und ist synchron in vivo. Dieser Prozess ist wenig untersucht worden. Die Hindernisse sind: (I) Freisetzung der Merozoiten findet in einer kurzen Zeit statt (<60 s) (II) Das Schizonten-Stadium der Parasiten hat eine hohe Empfindlichkeit gegenüber den Labor Bedingungen: pH-Wert und Zusammensetzung des Mediums, osmotischer Druck, Gas-Atmosphäre und Temperatur. (III) Die Erhaltung des Schizonten-Stadiums unter dem Mikroskop ist problematisch, so dass die Freisetzung der Merozoiten schwierig in Echtzeit zu beobachten ist. Das Ziel des Projektes war, die Freisetzung von Merozoiten von Plasmodium falciparum aus Erythrozyten mit der Schwingquarzbiosensor-Technik zu studieren. Dabei wurde die Änderung der Frequenz, aufgrund der Massenänderung bei der Freisetzung von Merozoiten, und auf einem zweiten Quarz die Reinfektion der nicht infizierten Erythrozyten durch die freigesetzten Merozoiten in Echtzeit beobachtet. Unsere SQ-Experimente beinhalten die folgenden Schritte: (I) Anpassung und Optimierung der Immobilisierung von biologischen Schichten, um Erythrozyten auf dem Quarz zu binden. (II) Optimierung von Bedingungen der Parasiten-Kultur in einer Schwingquarz-Messzelle. (III) Beobachtung des Frequenzsignals für infizierte und nicht-infizierte Erythrozyten und die Korrelation des Signals mit der Freisetzung von Merozoiten. (IV) Reinfektion gesunder Erythrozyten auf einem zweiten Quarz innerhalb eines SQ-System. (V) Test der Hemmung der Freisetzung der Merozoiten und der Reinfektion durch Merozoiten unter Einwirkung von Malariamedikamenten. Die Ergebnisse zeigten einen signifikanten Anstieg von ~ 1000 Hz am SQ für infizierte Erythrozyten, im Vergleich zu einem SQ mit gesunden Erythrozyten, wo das Frequenzsignal stabil geblieben ist. Mikroskopische Beobachtung der Oberfläche des Quarzes zu unterschiedlichen Zeitpunkten während des Experiments (REM und optisch) zeigte eine Korrelation zwischen dem Anstieg der Frequenz und der Freisetzung von Merozoiten. An diesem Punkt sind etwas mehr als 80% der infizierten Erythrozyten auf dem Quarz an dem Release beteiligt. Reinfektion neuer Erythrozyten wurde auf einem zweiten Quarz beobachtet. pH-Wert des Systems (7.2), Temperatur (37 ° C + / -0,1), Durchfluss des Mediums (9 uL / min), Sterilität des Prozesses (BactAlert), Gas-Atmosphäre (5% O2, Rest N2) wurden eingerichtet, um die Entwicklung der Parasiten und ihr Überleben sicher zu stellen. Externe Kontrollen mitttels Durchflusszytometrie 24 Stunden nach der Reinfektion zeigten einen Prozentsatz von Parasitämie von >1% in den in situ infizierten Erythrozyten. Unsere Ergebnisse zeigen, dass die Schwingquarz-Technologie ein geeignetes und wichtiges neues Instrument ist, um die Biologie des Re-Invasions-Prozesses der Plasmodien aufzuklären

Izaña Atmospheric Research Center. Activity Report 2015-2016

This report is a summary of the many activities at the Izaña Atmospheric Research Center to the broader community. The combination of operational activities, research and development in state-of-the-art measurement techniques, calibration and validation and international cooperation encompass the vision of WMO to provide world leadership in expertise and international cooperation in weather, climate, hydrology and related environmental issues

Izaña Atmospheric Research Center. Activity Report 2019-2020

Editors: Emilio Cuevas, Celia Milford and Oksana Tarasova.[EN]The Izaña Atmospheric Research Center (IARC), which is part of the State Meteorological Agency of Spain (AEMET), is a site of excellence in atmospheric science. It manages four observatories in Tenerife including the high altitude Izaña Atmospheric Observatory. The Izaña Atmospheric Observatory was inaugurated in 1916 and since that date has carried out uninterrupted meteorological and climatological observations, contributing towards a unique 100-year record in 2016. This reports are a summary of the many activities at the Izaña Atmospheric Research Center to the broader community. The combination of operational activities, research and development in state-of-the-art measurement techniques, calibration and validation and international cooperation encompass the vision of WMO to provide world leadership in expertise and international cooperation in weather, climate, hydrology and related environmental issues.[ES]El Centro de Investigación Atmosférica de Izaña (CIAI), que forma parte de la Agencia Estatal de Meteorología de España (AEMET), representa un centro de excelencia en ciencias atmosféricas. Gestiona cuatro observatorios en Tenerife, incluido el Observatorio de Izaña de gran altitud, inaugurado en 1916 y que desde entonces ha realizado observaciones meteorológicas y climatológicas ininterrumpidas y se ha convertido en una estación centenaria de la OMM. Estos informes resumen las múltiples actividades llevadas a cabo por el Centro de Investigación Atmosférica de Izaña. El liderazgo del Centro en materia de investigación y desarrollo con respecto a las técnicas de medición, calibración y validación de última generación, así como la cooperación internacional, le han otorgado una reputación sobresaliente en lo que se refiere al tiempo, el clima, la hidrología y otros temas ambientales afines

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

1er. Coloquio de educación para el diseño en la sociedad 5.0

Las memorias del 1er. Coloquio de Educación para el Diseño en la Sociedad 5.0 debenser entendidas como un esfuerzo colectivo de la comunidad de académicos de la División de Ciencias y Artes para el Diseño, que pone de manifiesto los retos y oportunidades que enfrenta la educación en diseño en un contexto de cambio acelerado y rompimiento de paradigmas.El evento se realizó el pasado mes de mayo de 2018 y se recibieron más de 50 ponencias por parte de las profesoras y profesores de la División.Las experiencias y/o propuestas innovadoras en cuanto a procesos de enseñanza y aprendizaje que presentan los autores en cada uno de sus textos son una invitación a reflexionar sobre nuestra situación actual en la materia, y emprender acciones en la División para continuar brindando una educación de calidad en diseño a nuestras alumnas, alumnos y la sociedad.Adicionalmente, se organizaron tres conferencias magistrales sobre la situación actual de la educación en Diseño y de las Instituciones de Educación Superior, impartidas por el Mtro. Luis Sarale, profesor de la Universidad Nacional de Cuyo en Mendoza (Argentina), y Presidente en su momento, de la Red de Carreras de Diseño en Universidades Públicas Latinoamericanas (DISUR), el Dr. Romualdo López Zárate, Rector de la Unidad Azcapotzalco, así como del Mtro. Luis Antonio Rivera Díaz, Jefe de Departamento de Teoría y Procesos del Diseño de la División de la Ciencias de la Comunicación y Diseño, en la Unidad Cuajimalpa de nuestra institución.La publicación de estas memorias son un esfuerzo divisional, organizado desde la Coordinación de Docencia Divisional y la Coordinación de Tecnologías del Aprendizaje, del Conocimiento y la Comunicación, para contribuir a los objetivos planteados en el documento ACCIONES:Agenda CyAD2021, en particular al eje de Innovación Educativa. Es necesario impulsar a todos los niveles de la División espacios de discusión orientados a reflexionar sobre el presente y futuro en la educación del diseñador, que contribuya a mejorar la calidad de la docencia y favorezca al fortalecimiento de los procesos de enseñanza y aprendizaje.Finalmente, extiendo un amplio reconocimiento a todos los miembros de la División que hicieron posible este evento, así como a todos los ponentes y participantes por compartir su conocimiento para que la División sea cada día mejor

COVID-19 symptoms at hospital admission vary with age and sex: results from the ISARIC prospective multinational observational study

Background: The ISARIC prospective multinational observational study is the largest cohort of hospitalized patients with COVID-19. We present relationships of age, sex, and nationality to presenting symptoms. Methods: International, prospective observational study of 60 109 hospitalized symptomatic patients with laboratory-confirmed COVID-19 recruited from 43 countries between 30 January and 3 August 2020. Logistic regression was performed to evaluate relationships of age and sex to published COVID-19 case definitions and the most commonly reported symptoms. Results: ‘Typical’ symptoms of fever (69%), cough (68%) and shortness of breath (66%) were the most commonly reported. 92% of patients experienced at least one of these. Prevalence of typical symptoms was greatest in 30- to 60-year-olds (respectively 80, 79, 69%; at least one 95%). They were reported less frequently in children (≤ 18 years: 69, 48, 23; 85%), older adults (≥ 70 years: 61, 62, 65; 90%), and women (66, 66, 64; 90%; vs. men 71, 70, 67; 93%, each P &lt; 0.001). The most common atypical presentations under 60 years of age were nausea and vomiting and abdominal pain, and over 60 years was confusion. Regression models showed significant differences in symptoms with sex, age and country. Interpretation: This international collaboration has allowed us to report reliable symptom data from the largest cohort of patients admitted to hospital with COVID-19. Adults over 60 and children admitted to hospital with COVID-19 are less likely to present with typical symptoms. Nausea and vomiting are common atypical presentations under 30 years. Confusion is a frequent atypical presentation of COVID-19 in adults over 60 years. Women are less likely to experience typical symptoms than men

Auditing of Monitoring and Respiratory Support Equipment in a Level III-C Neonatal Intensive Care Unit

Background. Random safety audits (RSAs) are a safety tool but have not been widely used in hospitals. Objectives. To determine the frequency of proper use of equipment safety mechanisms in relation to monitoring and mechanical ventilation by performing RSAs. The study also determined whether factors related to the patient, time period, or characteristics of the area of admission influenced how the device safety systems were used. Methods. A prospective observational study was conducted in a level III-C Neonatal Intensive Care Unit (NICU) during 2012. 87 days were randomly selected. Appropriate overall use was defined when all evaluated variables were correctly programmed in the audited device. Results. A total of 383 monitor and ventilator audits were performed. The Kappa coefficient of interobserver agreement was 0.93. The rate of appropriate overall use of the monitors and respiratory support equipment was 33.68%. Significant differences were found with improved usage during weekends, OR 1.85 (1.12–3.06, p=0.01), and during the late shift (3 pm to 10 pm), OR 1.59 (1.03–2.4, p=0.03). Conclusions. Equipment safety systems of monitors and ventilators are not properly used. To improve patient safety, we should identify which alarms are really needed and where the difficulties lie for the correct alarm programming

Izaña Atmospheric Research Center. Activity Report 2012-2014

This report is a summary of the many activities at the Izaña Atmospheric Research Center to the broader community. The combination of operational activities, research and development in state-of-the-art measurement techniques, calibration and validation and international cooperation encompass the vision of WMO to provide world leadership in expertise and international cooperation in weather, climate, hydrology and related environmental issues