Stem cell-related "self-renewal" signature and high epidermal growth factor receptor expression associated with resistance to concomitant chemoradiotherapy in glioblastoma

PURPOSE: Glioblastomas are notorious for resistance to therapy, which has been attributed to DNA-repair proficiency, a multitude of deregulated molecular pathways, and, more recently, to the particular biologic behavior of tumor stem-like cells. Here, we aimed to identify molecular profiles specific for treatment resistance to the current standard of care of concomitant chemoradiotherapy with the alkylating agent temozolomide. PATIENTS AND METHODS: Gene expression profiles of 80 glioblastomas were interrogated for associations with resistance to therapy. Patients were treated within clinical trials testing the addition of concomitant and adjuvant temozolomide to radiotherapy. RESULTS: An expression signature dominated by HOX genes, which comprises Prominin-1 (CD133), emerged as a predictor for poor survival in patients treated with concomitant chemoradiotherapy (n = 42; hazard ratio = 2.69; 95% CI, 1.38 to 5.26; P = .004). This association could be validated in an independent data set. Provocatively, the HOX cluster was reminiscent of a "self-renewal" signature (P = .008; Gene Set Enrichment Analysis) recently characterized in a mouse leukemia model. The HOX signature and EGFR expression were independent prognostic factors in multivariate analysis, adjusted for the O-6-methylguanine-DNA methyltransferase (MGMT) methylation status, a known predictive factor for benefit from temozolomide, and age. Better outcome was associated with gene clusters characterizing features of tumor-host interaction including tumor vascularization and cell adhesion, and innate immune response. CONCLUSION: This study provides first clinical evidence for the implication of a "glioma stem cell" or "self-renewal" phenotype in treatment resistance of glioblastoma. Biologic mechanisms identified here to be relevant for resistance will guide future targeted therapies and respective marker development for individualized treatment and patient selection.

Retinoic acid signaling organizes endodermal organ specification along the entire antero-posterior axis

BACKGROUND: Endoderm organ primordia become specified between gastrulation and gut tube folding in Amniotes. Although the requirement for RA signaling for the development of a few individual endoderm organs has been established a systematic assessment of its activity along the entire antero-posterior axis has not been performed in this germ layer. METHODOLOGY/PRINCIPAL FINDINGS: RA is synthesized from gastrulation to somitogenesis in the mesoderm that is close to the developing gut tube. In the branchial arch region specific levels of RA signaling control organ boundaries. The most anterior endoderm forming the thyroid gland is specified in the absence of RA signaling. Increasing RA in anterior branchial arches results in thyroid primordium repression and the induction of more posterior markers such as branchial arch Hox genes. Conversely reducing RA signaling shifts Hox genes posteriorly in endoderm. These results imply that RA acts as a caudalizing factor in a graded manner in pharyngeal endoderm. Posterior foregut and midgut organ primordia also require RA, but exposing endoderm to additional RA is not sufficient to expand these primordia anteriorly. We show that in chick, in contrast to non-Amniotes, RA signaling is not only necessary during gastrulation, but also throughout gut tube folding during somitogenesis. Our results show that the induction of CdxA, a midgut marker, and pancreas induction require direct RA signaling in endoderm. Moreover, communication between CdxA(+) cells is necessary to maintain CdxA expression, therefore synchronizing the cells of the midgut primordium. We further show that the RA pathway acts synergistically with FGF4 in endoderm patterning rather than mediating FGF4 activity. CONCLUSIONS/SIGNIFICANCE: Our work establishes that retinoic acid (RA) signaling coordinates the position of different endoderm organs along the antero-posterior axis in chick embryos and could serve as a basis for the differentiation of specific endodermal organs from ES cells.

Forcing of stratospheric chemistry and dynamics during the Dalton Minimum

The response of atmospheric chemistry and dynamics to volcanic eruptions and to a decrease in solar activity during the Dalton Minimum is investigated with the fully coupled atmosphere–ocean chemistry general circulation model SOCOL-MPIOM (modeling tools for studies of SOlar Climate Ozone Links-Max Planck Institute Ocean Model) covering the time period 1780 to 1840 AD. We carried out several sensitivity ensemble experiments to separate the effects of (i) reduced solar ultra-violet (UV) irradiance, (ii) reduced solar visible and near infrared irradiance, (iii) enhanced galactic cosmic ray intensity as well as less intensive solar energetic proton events and auroral electron precipitation, and (iv) volcanic aerosols. The introduced changes of UV irradiance and volcanic aerosols significantly influence stratospheric dynamics in the early 19th century, whereas changes in the visible part of the spectrum and energetic particles have smaller effects. A reduction of UV irradiance by 15%, which represents the presently discussed highest estimate of UV irradiance change caused by solar activity changes, causes global ozone decrease below the stratopause reaching as much as 8% in the midlatitudes at 5 hPa and a significant stratospheric cooling of up to 2 °C in the mid-stratosphere and to 6 °C in the lower mesosphere. Changes in energetic particle precipitation lead only to minor changes in the yearly averaged temperature fields in the stratosphere. Volcanic aerosols heat the tropical lower stratosphere, allowing more water vapour to enter the tropical stratosphere, which, via HOx reactions, decreases upper stratospheric and mesospheric ozone by roughly 4%. Conversely, heterogeneous chemistry on aerosols reduces stratospheric NOx, leading to a 12% ozone increase in the tropics, whereas a decrease in ozone of up to 5% is found over Antarctica in boreal winter. The linear superposition of the different contributions is not equivalent to the response obtained in a simulation when all forcing factors are applied during the Dalton Minimum (DM) – this effect is especially well visible for NOx/NOy. Thus, this study also shows the non-linear behaviour of the coupled chemistry-climate system. Finally, we conclude that especially UV and volcanic eruptions dominate the changes in the ozone, temperature and dynamics while the NOx field is dominated by the energetic particle precipitation. Visible radiation changes have only very minor effects on both stratospheric dynamics and chemistry.

Impact of geomagnetic excursions on atmospheric chemistry and dynamics

Geomagnetic excursions, i.e. short periods in time with much weaker geomagnetic fields and substantial changes in the position of the geomagnetic pole, occurred repeatedly in the Earth's history, e.g. the Laschamp event about 41 kyr ago. Although the next such excursion is certain to come, little is known about the timing and possible consequences for the state of the atmosphere and the ecosystems. Here we use the global chemistry climate model SOCOL-MPIOM to simulate the effects of geomagnetic excursions on atmospheric ionization, chemistry and dynamics. Our simulations show significantly increased concentrations of nitrogen oxides (NOx) in the entire stratosphere, especially over Antarctica (+15%), due to enhanced ionization by galactic cosmic rays. Hydrogen oxides (HOx) are also produced in greater amounts (up to +40%) in the tropical and subtropical lower stratosphere, while their destruction by reactions with enhanced NOx prevails over the poles and in high altitudes (by −5%). Stratospheric ozone concentrations decrease globally above 20 km by 1–2% and at the northern hemispheric tropopause by up to 5% owing to the accelerated NOx-induced destruction. A 5% increase is found in the southern lower stratosphere and troposphere. In response to these changes in ozone and the concomitant changes in atmospheric heating rates, the Arctic vortex intensifies in boreal winter, while the Antarctic vortex weakens in austral winter and spring. Surface wind anomalies show significant intensification of the southern westerlies at their poleward edge during austral winter and a pronounced northward shift in spring. Major impacts on the global climate seem unlikely.

Steroidogenesis of the testis - new genes and pathways

Defects of androgen biosynthesis cause 46,XY disorder of sexual development (DSD). All steroids are produced from cholesterol and the early steps of steroidogenesis are common to mineralocorticoid, glucocorticoid and sex steroid production. Genetic mutations in enzymes and proteins supporting the early biosynthesis pathways cause adrenal insufficiency (AI), DSD and gonadal insufficiency. The classic androgen biosynthesis defects with AI are lipoid CAH, CYP11A1 and HSD3B2 deficiencies. Deficiency of CYP17A1 rarely causes AI, and HSD17B3 or SRD5A2 deficiencies only cause 46,XY DSD and gonadal insufficiency. All androgen biosynthesis depends on 17,20 lyase activity of CYP17A1 which is supported by P450 oxidoreductase (POR) and cytochrome b5 (CYB5). Therefore 46,XY DSD with apparent 17,20 lyase deficiency may be due to mutations in CYP17A1, POR or CYB5. Illustrated by patients harboring mutations in SRD5A2, normal development of the male external genitalia depends largely on dihydrotestosterone (DHT) which is converted from circulating testicular testosterone (T) through SRD5A2 in the genital skin. In the classic androgen biosynthetic pathway, T is produced from DHEA and androstenedione/-diol in the testis. However, recently found mutations in AKR1C2/4 genes in undervirilized 46,XY individuals have established a role for a novel, alternative, backdoor pathway for fetal testicular DHT synthesis. In this pathway, which has been first elucidated for the tammar wallaby pouch young, 17-hydroxyprogesterone is converted directly to DHT by 5α-3α reductive steps without going through the androgens of the classic pathway. Enzymes AKR1C2/4 catalyse the critical 3αHSD reductive reaction which feeds 17OH-DHP into the backdoor pathway. In conclusion, androgen production in the fetal testis seems to utilize two pathways but their exact interplay remains to be elucidated.

Does the (structural) context matter? Analysing individual and structural factors of members’ commitment to sports clubs

Introduction: Over the last decades, Swiss sports clubs have lost their "monopoly" in the market for sports-related services and increasingly are in competition with other sports providers. For many sport clubs long-term membership cannot be seen as a matter of course. Current research on sports clubs in Switzerland – as well as for other European countries – confirms the increasing difficulties in achieving long-term member commitment. Looking at recent findings of the Swiss sport clubs report (Lamprecht, Fischer & Stamm, 2012), it can be noted, that a decrease in memberships does not equally affect all clubs. There are sports clubs – because of their specific situational and structural conditions – that have few problems with member fluctuation, while other clubs show considerable declines in membership. Therefore, a clear understanding of individual and structural factors that trigger and sustain member commitment would help sports clubs to tackle this problem more effectively. This situation poses the question: What are the individual and structural determinants that influence the tendency to continue or to quit the membership? Methods: Existing research has extensively investigated the drivers of members’ commitment at an individual level. As commitment of members usually occurs within an organizational context, the characteristics of the organisation should be also considered. However, this context has been largely neglected in current research. This presentation addresses both the individual characteristics of members and the corresponding structural conditions of sports clubs resulting in a multi-level framework for the investigation of the factors of members’ commitment in sports clubs. The multilevel analysis grant a adequate handling of hierarchically structured data (e.g., Hox, 2002). The influences of both the individual and context level on the stability of memberships are estimated in multi-level models based on a sample of n = 1,434 sport club members from 36 sports clubs. Results: Results of these multi-level analyses indicate that commitment of members is not just an outcome of individual characteristics, such as strong identification with the club, positively perceived communication and cooperation, satisfaction with sports clubs’ offers, or voluntary engagement. It is also influenced by club-specific structural conditions: stable memberships are more probable in rural sports clubs, and in clubs that explicitly support sociability, whereas sporting-success oriented goals in clubs have a destabilizing effect. Discussion/Conclusion: The proposed multi-level framework and the multi-level analysis can open new perspectives for research concerning commitment of members to sports clubs and other topics and problems of sport organisation research, especially in assisting to understand individual behavior within organizational contexts. References: Hox, J. J. (2002). Multilevel analysis: Techniques and applications. Mahwah: Lawrence Erlbaum. Lamprecht, M., Fischer, A., & Stamm, H.-P. (2012). Die Schweizer Sportvereine – Strukturen, Leistungen, Herausforderungen. Zurich: Seismo.

"Does the context matter?” Individuelle und strukturelle Faktoren der Mitgliederbindung in Sportvereinen

Einleitung Folgt man den aktuellen Sportentwicklungsberichten, dann sehen sich zunehmend mehr Sportvereine mit Mitgliederfluktuationen sowie stagnierenden bzw. zurückgehenden Mitglie- derzahlen konfrontiert (Lamprecht et al. 2012). Jedoch werden nicht alle Vereine in gleichem Maße mit instabilen Mitgliedschaftsverhältnissen konfrontiert. So gibt es mit Blick auf die Mitgliederstruktur Vereine, die – aufgrund ihrer spezifischen situativen und strukturellen Bedingungen – kaum Probleme mit Mitgliederfluktuation und Vereinsaustritten haben, wohin- gegen andere Vereine mitunter erhebliche Mitgliederrückgänge verzeichnen. Demnach ist zu vermuten, dass sich das soziale Handeln der Vereinsmitglieder je nach Organisationsprofil der Vereine unterscheidet. Zwar werden Verknüpfungen von Individual- und korrespondierenden Strukturdaten innerhalb der Sportvereinsforschung bereits seit geraumer Zeit gefordert (z.B. Nagel, 2007), aber bis heute nicht konsequent umgesetzt. Es stellt sich deshalb die Frage, welche organisations- und individuumsbezogenen Faktoren für die Mitgliederbindung in Sportvereinen eine Rolle spielen? Theoretisch-methodisches Vorgehen Im Zusammenhang mit der Frage der Mitgliederbindung wird davon ausgegangen, dass kontextuelle Bedingungen individuelle Entscheidungen strukturieren und somit persönliche Handlungsketten beeinflussen können (Coleman, 1990). Auf dieser Grundlage wird ein Mehr- ebenenmodell entwickelt, das neben individuellen Merkmalen auch die Strukturbedingungen von Sportvereinen berücksichtigt, die im Zusammenhang mit der individuellen Wahlhandlung zwischen stabiler Mitgliedschaft oder Austritt stehen. Der organisationale Kontext Sportverein wird dabei als Interessenorganisation konzeptualisiert, der mit seinen Kontexteigenschaften als Gelegenheits- und Opportunitätsstruktur, als kultureller sowie sozialer Bezugsrahmen gewisse Anreize schafft („Logik der Situation“), die gemäss individueller Präferenzen („Logik der Selektion“) zu Parametern des Mitgliederhandelns werden können. Die aus dem Modell abgeleiteten Annahmen werden auf der Grundlage einer Mitglieder- befragung (n = 1.434) in 36 Schweizer Sportvereinen empirisch geprüft. Die adäquate Methode, welche die hierarchische Datenstruktur (jede Messung auf der Individualebene kann eindeutig einer Messung auf der Vereinsebene zugeordnet werden) adäquat berücksichtigt und folglich das entwickelte theoretische Modell statistisch umsetzt, ist die Mehrebenenanalyse (z.B. Hox, 2002). Entsprechend wird der Einfluss der Individual- und Kontextebene auf die Mitglieder- bindung in Sportvereinen anhand unterschiedlicher Mehrebenenmodelle (Random Intercept, Random Slope sowie Cross-Level Interaktionen) geschätzt. Ergebnisse Die Analysen machen deutlich, dass sich die dauerhafte Mitgliedschaft in Sportvereinen nicht allein auf individuelle Merkmale der Mitgliedschaft, wie eine ausgeprägte Verbundenheit, ein positiv wahrgenommenes soziales Miteinander, die Zufriedenheit mit der Vereinsarbeit sowie die ehrenamtliche Mitarbeit zurückführen lässt. Darüber hinaus nehmen auch vereinsspezi-fische Strukturbedingungen Einfluss auf die Mitgliederbindung, wobei in ländlich geprägten Sportvereinen und in Vereinen, die Geselligkeit explizit fordern und in denen das Vereinsziel sportlicher Erfolg eher eine untergeordnete Rolle spielt, die Austrittswahrscheinlichkeit geringer ist. Diskussion Die Befunde machen deutlich, dass für eine dauerhafte Mitgliedschaft sowohl zweckorientierte Nutzenüberlegungen als auch solidargemeinschaftliche Handlungsorientierungen eine zentrale Rolle spielen, so dass eine ausschließliche Dienstleistungs- bzw. Kundenorientierung als Strategie der Mitgliederbindung in Sportvereinen, wie sie vielfach (auch von Verbänden) nahegelegt wird, zu kurz greifen dürfte. Weiterhin zeigt sich, dass der Sportverein als Ort der Geselligkeit nicht nur Werte des sozialen Miteinanders und solidarischen Verhaltens vermittelt, sondern auch als Katalysator der Stabilität der Mitgliedschaft wirkt, sofern entsprechende Gelegenheiten zur Verfügung stehen. Im Zusammenhang mit der Mitgliederbindung scheint damit gerade jene vereinskulturelle Orientierung von Vereinen bedeutsam, die im Zuge der Modernisierung von Vereinsangeboten gern als überholt erachtet wird. Literatur Coleman, J. S. (1990). Foundations of social theory. Cambridge, MA: Belknap. Hox, J. (2002). Multilevel analysis. Techniques and applications. Mahwah: Erlbaum. Lamprecht, M., Fischer, A. & Stamm, H.-P. (2012). Die Schweizer Sportvereine – Strukturen, Leistungen, Herausforderungen. Zürich: Seismo. Nagel, S. (2007). Akteurtheoretische Analyse der Sportvereinsentwicklung – ein theoretisch- methodischer Bezugsrahmen. Sportwissenschaft, 37, 186–201.

Ectopic Meis1 expression in the mouse limb bud alters P-D patterning in a Pbx1-independent manner

During limb development, expression of the TALE homeobox transcription factor Meis1 is activated by retinoic acid in the proximal-most limb bud regions, which give rise to the upper forelimb and hindlimb. Early subdivision of the limb bud into proximal Meis-positive and distal Meis-negative domains is necessary for correct proximo-distal (P-D) limb development in the chick, since ectopic Meis1 overexpression abolishes distal limb structures, produces a proximal shift of limb identities along the P-D axis, and proximalizes distal limb cell affinity properties. To determine whether Meis activity is also required for P-D limb specification in mammals, we generated transgenic mice ectopically expressing Meis1 in the distal limb mesenchyme under the control of the Msx2 promoter. Msx2:Meis1 transgenic mice display altered P-D patterning and shifted P-D Hox gene expression domains, similar to those previously described for the chicken. Meis proteins function in cooperation with PBX factors, another TALE homeodomain subfamily. Meis-Pbx interaction is required for nuclear localization of both proteins in cell culture, and is important for their DNA-binding and transactivation efficiency. During limb development, Pbx1 nuclear expression correlates with the Meis expression domain, and Pbx1 has been proposed as the main Meis partner in this context; however, we found that Pbx1 deficiency did not modify the limb phenotype of Msx2:Meis1 mice. Our results indicate a conserved role of Meis activity in P-D specification of the tetrapod limb and suggest that Pbx function in this context is either not required or is provided by partners other than Pbx1.