Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes

Research in autophagy continues to accelerate,(1) and as a result many new scientists are entering the field. Accordingly, it is important to establish a standard set of criteria for monitoring macroautophagy in different organisms. Recent reviews have described the range of assays that have been used for this purpose.(2,3) There are many useful and convenient methods that can be used to monitor macroautophagy in yeast, but relatively few in other model systems, and there is much confusion regarding acceptable methods to measure macroautophagy in higher eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers of autophagosomes versus those that measure flux through the autophagy pathway; thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from fully functional autophagy that includes delivery to, and degradation within, lysosomes (in most higher eukaryotes) or the vacuole (in plants and fungi). Here, we present a set of guidelines for the selection and interpretation of the methods that can be used by investigators who are attempting to examine macroautophagy and related processes, as well as by reviewers who need to provide realistic and reasonable critiques of papers that investigate these processes. This set of guidelines is not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to verify an autophagic response.

Identification of novel mutations in X-linked retinitis pigmentosa families and implications for diagnostic testing

PURPOSE: The goal of this study was to identify mutations in X-chromosomal genes associated with retinitis pigmentosa (RP) in patients from Germany, The Netherlands, Denmark, and Switzerland. METHODS: In addition to all coding exons of RP2, exons 1 through 15, 9a, ORF15, 15a and 15b of RPGR were screened for mutations. PCR products were amplified from genomic DNA extracted from blood samples and analyzed by direct sequencing. In one family with apparently dominant inheritance of RP, linkage analysis identified an interval on the X chromosome containing RPGR, and mutation screening revealed a pathogenic variant in this gene. Patients of this family were examined clinically and by X-inactivation studies. RESULTS: This study included 141 RP families with possible X-chromosomal inheritance. In total, we identified 46 families with pathogenic sequence alterations in RPGR and RP2, of which 17 mutations have not been described previously. Two of the novel mutations represent the most 3'-terminal pathogenic sequence variants in RPGR and RP2 reported to date. In exon ORF15 of RPGR, we found eight novel and 14 known mutations. All lead to a disruption of open reading frame. Of the families with suggested X-chromosomal inheritance, 35% showed mutations in ORF15. In addition, we found five novel mutations in other exons of RPGR and four in RP2. Deletions in ORF15 of RPGR were identified in three families in which female carriers showed variable manifestation of the phenotype. Furthermore, an ORF15 mutation was found in an RP patient who additionally carries a 6.4 kbp deletion downstream of the coding region of exon ORF15. We did not identify mutations in 39 sporadic male cases from Switzerland. CONCLUSIONS: RPGR mutations were confirmed to be the most frequent cause of RP in families with an X-chromosomal inheritance pattern. We propose a screening strategy to provide molecular diagnostics in these families.

Steuer-, Devisen- und Einziehungsakten als neue Quellen der Zeitgeschichtsforschung

Die Unterlagen der Finanzverwaltung sind in den letzten Jahren verstärkt als neue Quellengattung in das Blickfeld zeithistorischer Forschungen geraten. Für den Themenkomplex 'Raub und Wiedergutmachung' sind die zeitgenössischen Quellen besonders der Jahre 1933 bis 1945 vor allem wegen ihrer Unmittelbarkeit von großer Bedeutung. Zur verstärkten Nutzung der Finanzakten in der Forschung hat nicht zuletzt die liberaler gehandhabte Archivbenützungspraxis entscheidend beigetragen, die deswegen in diesem Beitrag näher erläutert wird. Zudem werden exemplarisch drei Quellengruppen aus dem Bereich der bayerischen Finanzverwaltung näher beleuchtet: Steuer-, Devisen- und Einziehungsakten. Sie dokumentieren den schrittweisen Vermögensentzug der jüdischen Bevölkerung von der Erfassung über die Entziehung bis zur Verwertung aus der Täterperspektive und werden hier nach Inhalt, Quantität, Aufbau und Aussagewert analysiert. Der Beitrag konzentriert sich dabei auf die Überlieferung dieser Akten in den staatlichen Archiven Bayerns.

Short-term feed intake is regulated by macronutrient oxidation in lactating Holstein cows

In addition to plasma metabolites and hormones participating as humoral signals in the control of feed intake, oxidative metabolic processes in peripheral organs also generate signals to terminate feeding. Although the degree of oxidation over longer periods is relatively constant, recent work suggests that the periprandial pattern of fuel oxidation is involved in regulating feeding behavior in the bovine. However, the association between periprandial oxidative metabolism and feed intake of dairy cows has not yet been studied. Therefore, the aim of this study was to elucidate possible associations existing between single feed intake events and whole-body net fat and net carbohydrate oxidation as well as their relation to plasma metabolite concentrations. To this end, 4 late-lactating cows equipped with jugular catheters were kept in respiratory chambers with continuous and simultaneous recording of gas exchange and feed intake. Animals were fed ad libitum (AL) for 24h and then feed restricted (RE) to 50% of the previous AL intake for a further 24h. Blood samples were collected hourly to analyze β-hydroxybutyrate (BHBA), glucose, nonesterified fatty acids (NEFA), insulin, and acylated ghrelin concentrations. Cross-correlation analysis revealed an offset ranging between 30 and 42 min between the maximum of a feed intake event and the lowest level of postprandial net fat oxidation (FOX(net)) and the maximum level of postprandial net carbohydrate oxidation (COX(net)), respectively. During the AL period, FOX(net) did not increase above -0.2g/min, whereas COX(net) did not decrease below 6g/min before the start of the next feed intake event. A strong inverse cross-correlation was obtained between COX(net) and plasma glucose concentration. Direct cross-correlations were observed between COXnet and insulin, between heat production and BHBA, between insulin and glucose, and between BHBA and ghrelin. We found no cross-correlation between FOX(net) and NEFA. During RE, FOX(net) increased with an exponential slope, exceeded the threshold of -0.2g/min as indicated by increasing plasma NEFA concentrations, and approached a maximum rate of 0.1g/min, whereas COX(net) decayed in an exponential manner, approaching a minimal COX(net) rate of about 2.5 g/min in all cows. Our novel findings suggest that, in late-lactating cows, postprandial increases in metabolic oxidative processes seem to signal suppression of feed intake, whereas preprandially an accelerated FOX(net) rate and a decelerated COX(net) rate initiate feed intake.

Erwerbsbeteiligung und Berufschancen von Frauen. Theoretische Ansätze, methodische Verfahren und empirische Ergebnisse aus der Lebensverlaufsperspektive

Mit Hilfe der Daten der Lebensverlaufsstudie des Max-Planck-Instituts für Bildungsforschung gehen die Autoren der Frage nach, welche arbeitsmarktbezogenen und institutionellen Faktoren die Erwerbsbeteiligung von Frauen im Lebenslauf beeinflussen und inwieweit sich für unterschiedliche Geburtenkohorten neue Erwerbsbeteiligungsmuster etablieren. Die zugrundliegenden dynamischen Betrachtungen beziehen die Wirkungen vorausgegangener Ereignisse auf die zukünftigen Verlaufsstrukturen mit ein, berücksichtigen bedeutsame Strukturmerkmale für zentrale Lebensbedingungen und versuchen Interdependenzen verschiedener gesellschaftlicher Bereiche zu integrieren. Das Eintreten von für den Lebens- und Berufsverlauf von Frauen typischen Ereignissen wird mit Survival-Analysen vorausgeschätzt.

Estilos en las redes sociales en línea: un estudio basado en corpora

Las nuevas tecnologías y el procesamiento digital han facilitado considerablemente la lingüística de corpus, por ejemplo Internet es una herramienta fácil y barata para recopilar corpus. Internet es cada vez más popular y más importante para la comunicación a causa de la enorme influencia de los nuevos medios y ha afectado la vida y la sociedad de muchas maneras y en parte, de manera fundamental. No sorprende por eso que la lengua y la comunicación misma se vean afectadas. Uno de los fenómenos más interesantes dentro de la comunicación mediada por ordenadores (CMC) son las redes sociales en línea, que en pocos años se han convertido en un medio de comunicación muy difundido y en expansión continua. Su estudio es particularmente interesante porque debido al desarrollo constante de la tecnología las redes sociales en línea no son una entidad estática, sino que cambian incesantemente, introduciéndose frecuentemente novedades para su uso. Estas novedades están condicionadas por el medio electrónico que a su vez influye decisivamente en el estilo de comunicación empleado en redes sociales como Facebook y Tuenti. Al ser un nuevo medio de interacción social, las redes sociales en línea producen un estilo de comunicación propio. El objetivo de análisis de mi tesis es cómo los usuarios de Facebook y Tuenti de la ciudad de Málaga crean este estilo mediante el uso de rasgos fónicos propios de la variedad andaluza y de qué manera la actitud lingüística de los usuarios influye en el uso de dichos rasgos fónicos. Este estudio se basa en un corpus elaborado a partir de enunciados de informantes en Facebook y Tuenti. Un corpus constituido por transcripciones amplias de grabaciones de hablantes malagueños me sirve de corpus de comparación. Otra herramienta metodológica empleada para recopilar datos será la encuesta: un tipo de encuesta estará destinada a captar las actitudes de los participantes frente a diversos rasgos del habla andaluza/malagueña y otro a examinar por qué la gente utiliza estos rasgos en Facebook y Tuenti. Este estudio se apoya en los resultados de un estudio piloto que muestran que los factores sociales y lingüísticos analizados funcionan de manera distinta en el habla real y virtual. Debido a estos usos diferentes podemos considerar la comunicación electrónica de Facebook y Tuenti como un estilo condicionado por el tipo de espacio virtual. Se trata de un estilo que sirve a los usuarios para crear significado social y para expresar sus identidades a partir de la lengua.