Single cell analysis of Vibrio harveyi uncovers functional heterogeneity in response to quorum sensing signals

Background: Vibrio harveyi and closely related species are important pathogens in aquaculture. A complex quorum sensing cascade involving three autoinducers controls bioluminescence and several genes encoding virulence factors. Single cell analysis of a V. harveyi population has already indicated intercellular heterogeneity in the production of bioluminescence. This study was undertaken to analyze the expression of various autoinducer-dependent genes in individual cells. Results: Here we used reporter strains bearing promoter::gfp fusions to monitor the induction/repression of three autoinducer-regulated genes in wild type conjugates at the single cell level. Two genes involved in pathogenesis - vhp and vscP, which code for an exoprotease and a component of the type III secretion system, respectively, and luxC (the first gene in the lux operon) were chosen for analysis. The lux operon and the exoprotease gene are induced, while vscP is repressed at high cell density. As controls luxS and recA, whose expression is not dependent on autoinducers, were examined. The responses of the promoter:: gfp fusions in individual cells from the same culture ranged from no to high induction. Importantly, simultaneous analysis of two autoinducer induced phenotypes, bioluminescence (light detection) and exoproteolytic activity (fluorescence of a promoter:: gfp fusion), in single cells provided evidence for functional heterogeneity within a V. harveyi population. Conclusions: Autoinducers are not only an indicator for cell density, but play a pivotal role in the coordination of physiological activities within the population

Playful training of numerical skills in kindergarten for prevention of mathematical disability

In der vorliegenden Evaluationsstudie wurde untersucht, ob numerische Kompetenzen im Vorschulalter durch in den Kindergartenalltag integrierte zahlen- und mengenbezogene Spiele gefördert werden können. Darüber hinaus war es von Interesse herauszufinden, ob Kindergartenkinder mit weniger gut entwickelten numerischen Kompetenzen insofern von dieser Fördermethode profitieren, als dass sie ihren Leistungsrückstand aufholen können. Realisiert wurde ein Prätest-Posttest-Design mit einer Fördergruppe, einer Kontrollgruppe mit Kontrollintervention sowie einer Wartekontrollgruppe. Insgesamt nahmen 142 Kinder im Alter von vier bis fünf Jahren an der Studie teil. Die Ergebnisse kovarianzanalytischer Verfahren mit Messwiederholung sprechen dafür, dass sich die numerischen Kompetenzen von Kindergartenkindern anhand des verwendeten Förderkonzepts steigern lassen. Außerdem konnte gezeigt werden, dass jene Kinder der Fördergruppe, deren numerische Leistung zum Prätestzeitpunkt unter dem Median der Gesamtstichprobe lag, einen höheren Leistungszuwachs aufwiesen als die leistungsstärkeren Kinder, die nicht mit den zahl- und mengenbezogenen Spielen gefördert wurden. Die Ergebnisse weisen somit auch auf eine Eignung des spielerischen Förderkonzepts zur Kompensation von Entwicklungsnachteilen bezüglich der numerischen Kompetenz hin. (DIPF/Orig.)In this study, we investigated if preschool children´s numerical skills can be fostered by playing number- and quantity-related games during children´s daily playtimes. Furthermore, we wanted to find out if preschoolers with poorly developed numerical skills catch up on their developmental delay by playing these games. In order to do so, we used a pretest-posttest-design with (a) an intervention group playing number- and quantity-related games, (b) a control group participating in another intervention with the same basic conditions but other intervention goals and (c) a control group without an intervention. One hundred forty-two children aged 4 to 5 years participated in the study. The results show that preschoolers’ numerical skills can be increased by playing number- and quantity-related games. Furthermore, the numerical skills of children with poorly developed numerical skills increased more significantly than the numerical skills of children with better developed numerical skills who did not play the number- and quantity-related games. Thus, results indicate that playing these number- and quantity-related games might be used to compensate for early developmental disadvantage in numerical skills. (DIPF/Orig.

Towards transnational interoperable PPDR communications: the European ISI cloud network

The European Council has been stressing the need for interoperability among technologies used for Public Protection and Disaster Relief (PPDR) communications across Europe for a long time. Nevertheless, while the introduction of TETRA and TETRAPOL technologies in the last two decades has increased the possibility to talk cross agency internally in a country, cross border communication for the public safety forces is not well solved as of today. This paper describes the communications interoperability solution that is being developed in the framework of the ISITEP project. This solution, referred to as the European Inter-System Interface (ISI) Cloud Network, aims to integrate the PPDR national/regional infrastructures to allow migration (i.e., roaming) and communication services between networks within a secure framework. The ISI Cloud Network involves, among other components, the specification of a new ISI interface to be deployed over IP transport networks and the development of a number of different gateways to cover the use of TETRA and TETRAPOL technologies as well as the use of legacy TETRA ISI by some networks.Peer ReviewedPostprint (author's final draft

Social Policy Responses of the Chilean State to the Earthquake and Tsunami of 2010

This is the author accepted manuscript. The final version is available from SAGE via https://doi.org/10.1177/0094582X16648955 Decades of neoliberal policy have left Chile with a skeletal state that administers social policy through targeting and outsourcing in public-private partnerships that lack coordination. The reconstruction after the 2010 earthquake and tsunami responded to the emergency largely according to these same principles. While official reports on the reconstruction effort show a state that is complying with its goals, evidence from fieldwork in the city of Constitución illustrates that this method is highly inadequate in the context of a natural disaster. Chile should establish a social policy structure for natural disasters that allows for a rapid response to a social emergency based on universal or near-universal allocation criteria. Varias décadas de políticas neoliberales han dejado a Chile con un estado débil que simplemente administra las políticas sociales por medio de la focalización y las subcontrataciones en alianzas público-privadas que carecen de coordinación. La reconstrucción después del terremoto y el sunami de 2010 respondió a la catástrofe mayormente de acuerdo a estos mismos principios. Mientras los informes oficiales sobre los esfuerzos de reconstrucción muestran a un estado que cumple con sus metas, el trabajo de campo en la ciudad de Constitución demuestra que este método es decididamente insuficiente en el contexto de un desastre natural. Chile debe establecer unas estructuras de política pública para lidiar con los desastres naturales que permitan dar una respuesta rápida a las emergencias sociales en base a criterios de asignación universales o casi universales. </jats:p

Styles of leadership, fears of compassion, and competing to avoid inferiority.

There is general agreement that styles of leadership evolved from mammalian group living strategies that form social ranks. In both non-human primates and humans, different styles of hierarchical dominant-subordinate and leader-follower behavior can be observed. These can be described in terms of dimensions of antisocial (relatively self-focused, aggressive and threat-based) and prosocial (relatively empathic, caring, and supportive) interpersonal styles. The aim of this study was to explore how a set of established self-report questionnaires might relate to these two dimensions. Two hundred and nineteen students completed questionnaires assessing ruthless self-advancement, coalition building, and dominant leadership styles, as well as hypercompetitiveness, narcissism, striving to avoid inferiority, compassion focused and ego focused goals, fears of compassion, social safeness and attachment (in)security. A principal component analysis supported an antisocial leadership style factor which comprised of ruthless self-advancement, narcissism and hypercompetitiveness. This was significantly correlated with fears of compassion, ego focused goals, insecure striving (striving to avoid inferiority), fears of losing out, fears of being overlooked, fears of being rejected, and avoidant relating in close relationships. It was significantly negatively correlated with compassionate goals. As the results did not reveal a clear factor solution for a prosocial leadership style, we chose to use the coalition building leadership style variable. This showed the opposite pattern, being significantly negatively correlated with narcissism, hypercompetitiveness, fears of compassion, fears of active rejection, and avoidance in close relationships. It was significantly positively correlated with secure striving, compassionate goals, and social safeness. We also found that fears of compassion for others was a partial mediator of the relationship between insecure striving with antisocial leadership style. Moreover, lower fears of compassion for the self emerged as a key mediator for the relationship between non-avoidant attachment with coalition building leadership style and, secure non-striving with coalition building leadership style. While the motive to accumulate social power, resources and dominance may be linked to antisocial forms of leadership, the intensity of the drive may also be linked to unaddressed threats and fears of rejection and fears of compassion. Efforts to promote more ethical, moral and prosocial forms of leadership may falter if such fears are left unaddressed.This research was supported by a donation received by the Compassionate Mind Foundation charity (https://compassionatemind.co.uk/) from Slimming World

Sex determining region Y-box 2 (SOX2) amplification is an independent indicator of disease recurrence in sinonasal cancer.

The transcription factor SOX2 (3q26.3-q27) is an embryonic stem cell factor contributing to the induction of pluripotency in terminally differentiated somatic cells. Recently, amplification of the SOX2 gene locus has been described in squamous cell carcinoma (SCC) of different organ sites. Aim of this study was to investigate amplification and expression status of SOX2 in sinonasal carcinomas and to correlate the results with clinico-pathological data. A total of 119 primary tumor samples from the sinonasal region were assessed by fluorescence in-situ hybridization and immunohistochemistry for SOX2 gene amplification and protein expression, respectively. Of these, 59 were SSCs, 18 sinonasal undifferentiated carcinomas (SNUC), 10 carcinomas associated with an inverted papilloma (INVC), 19 adenocarcinomas (AD) and 13 adenoid cystic carcinomas (ACC). SOX2 amplifications were found in subsets of SCCs (37.5%), SNUCs (35.3%), INVCs (37.5%) and ADs (8.3%) but not in ACCs. SOX2 amplification resulted in increased protein expression. Patients with SOX2-amplified sinonasal carcinomas showed a significantly higher rate of tumor recurrences than SOX2 non-amplified tumors. This is the first study assessing SOX2 amplification and expression in a large cohort of sinonasal carcinomas. As opposed to AD and ACC, SOX2 amplifications were detected in more than 1/3 of all SCCs, SNUCs and INVCs. We therefore suggest that SNUCs are molecularly closely related to SCCs and INVCs and that these entities represent a subgroup of sinonasal carcinomas relying on SOX2 acquisition during oncogenesis. SOX2 amplification appears to identify sinonasal carcinomas that are more likely to relapse after primary therapy, suggesting that these patients might benefit from a more aggressive therapy regime

Identification of late assembly domains of the human endogenous retrovirus-K(HML-2)

Background: Late assembly (L)-domains are protein interaction motifs, whose dysfunction causes characteristic budding defects in enveloped viruses. Three different amino acid motifs, namely PT/SAP, PPXY and YPXnL have been shown to play a major role in the release of exogenous retroviruses. Although the L-domains of exogenous retroviruses have been studied comprehensively, little is known about these motifs in endogenous human retroviruses. Results: Using a molecular clone of the human endogenous retrovirus K113 that had been engineered to reverse the presumed non-synonymous postinsertional mutations in the major genes, we identified three functional L-domains of the virus, all located in the Gag p15 protein. A consensus PTAP tetrapeptide serves as the core of a main L-domain for the virus and its inactivation reduces virus release in HEK 293T cells by over 80%. Electron microscopy of cells expressing the PTAP mutant revealed predominantly late budding structures and budding chains at the plasma membrane. The fact that this motif determines subcellular colocalization with Tsg101, an ESCRT-I complex protein known to bind to the core tetrapeptide, supports its role as an L-domain. Moreover, two YPXnL motifs providing additional L-domain function were identified in the p15 protein. One is adjacent to the PTAP sequence and the other is in the p15 N-terminus. Mutations in either motif diminishes virus release and induces an L-domain phenotype while inactivation of all three L-domains results in a complete loss of particle release in HEK 293T cells. The flexibility of the virus in the use of L-domains for gaining access to the ESCRT machinery is demonstrated by overexpression of Tsg101 which rescues the release of the YPXnL mutants. Similarly, overexpression of Alix not only enhances release of the PTAP mutant by a factor of four but also the release of a triple mutant, indicating that additional cryptic YPXnL domains with a low affinity for Alix may be present. No L-domain activity is provided by the proline-rich peptides at the Gag C-terminus. Conclusions: Our data demonstrate that HERV-K(HML-2) release is predominantly mediated through a consensus PTAP motif and two auxiliary YPXnL motifs in the p15 protein of the Gag precursor

Exploring how family carers of a person with dementia manage pre-death grief: A mixed methods study

Objectives: Many family carers of a person with dementia experience pre-death grief. We aimed to identify strategies that help carers manage pre-death grief. We hypothesised that emotion and problem focussed styles would be associated with lower, and dysfunctional coping with higher grief intensity. Methods: Mixed methods observational study using structured and semi-structured interviews with 150 family carers of people with dementia living at home or in a care home. Most participants were female (77%), caring for a parent (48%) or partner/spouse (47%) with mild (25%), moderate (43%) or severe (32%) dementia. They completed the Marwit-Meuser Caregiver Grief Inventory Short Form and the Brief Coping Orientation to Problems Experienced (Brief-COPE) questionnaire. We asked carers to identify strategies used for managing grief. We recorded field notes for 150 interviews and audio-recorded additional interviews with a sub-sample of 16 participants. Results: Correlations indicated that emotion-oriented coping was associated with lower grief (R = −0.341), and dysfunctional coping with higher grief (R = 0.435), with a small association with problem-focused strategies (R = −0.109), partly supporting our hypothesis. Our qualitative themes broadly match the three Brief-COPE styles. Unhelpful strategies of denial and avoidance align with dysfunctional coping strategies. Psychological strategies (including acceptance and humour) and seeking support were consistent with emotion-focused strategies, but we did not identify a theme relating to problem-focused strategies. Conclusion: Most carers identified multiple strategies for processing grief. Carers could readily identify supports and services that they found helpful for managing pre-death grief, yet current services appear under-resourced to meet growing demand. (ClinicalTrials.gov ID: NCT03332979)