Preferential expansion and survival of B lymphocytes based on VH framework 1 and framework 3 expression: "positive" selection in appendix of normal and VH-mutant rabbits.

B cells with a rearranged heavy-chain variable region VHa allotype-encoding VH1 gene segment predominate throughout the life of normal rabbits and appear to be the source of the majority of serum immunoglobulins, which thus bear VHa allotypes. The functional role(s) of these VH framework region (FR) allotypic structures has not been defined. We show here that B cells expressing surface immunoglobulin with VHa2 allotypic specificities are preferentially expanded and positively selected in the appendix of young rabbits. By flow cytometry, a higher proportion of a2+ B cells were progressing through the cell cycle (S/G2/M) compared to a2- B cells, most of which were in the G1/G0 phase of the cell cycle. The majority of appendix B cells in dark zones of germinal centers of normal 6-week-old rabbits were proliferating and very little apoptosis were observed. In contrast, in 6-week-old VH-mutant ali/ali rabbits, little cell proliferation and extensive apoptosis were observed. Nonetheless even in the absence of VH1, B cells with a2-like surface immunoglobulin had developed and expanded in the appendix of 11-week-old mutants. The numbers and tissue localization of B cells undergoing apoptosis then appeared similar to those found in 6-week-old normal appendix. Thus, B cells with immunoglobulin receptors lacking the VHa2 allotypic structures were less likely to undergo clonal expansion and maturation. These data suggest that "positive" selection of B lymphocytes through FR1 and FR3 VHa allotypic structures occurs during their development in the appendix.

Dual transcriptional control by Ear3/COUP: negative regulation through the DR1 direct repeat and positive regulation through a sequence downstream of the transcriptional start site of the mouse mammary tumor virus promoter.

Ear3/COUP is an orphan member of the steroid/thyroid hormone receptor superfamily of transcription factors and binds most tightly to a direct repeat of AGGTCA with 1 nucleotide in between (DR1). Ear3/COUP also binds with a similar affinity to the palindromic thyroid hormone response element (TRE). This binding preference of Ear3/COUP is same as that of the retinoid X receptor (RXR), which is another member of the superfamily. In the present study, we identified a sequence responsible for Ear3/COUP-mediated transactivation in the region downstream of the transcription start site of the mouse mammary tumor virus promoter. This cis-acting sequence was unresponsive to RXR. When the DR1 or TRE sequence was added upstream of the promoter, transactivation by Ear3/COUP was completely abolished, whereas RXR enhanced transcription from the promoter. The mode of action of Ear3/COUP could be utilized to control complex gene expressions in morphogenesis, homeostasis, and development.

Innovative analytical strategies for the development of sensor devices and mass spectrometry methods

Il lavoro presentato in questa tesi di Dottorato è incentrato sullo sviluppo di strategie analitiche innovative basate sulla sensoristica e su tecniche di spettrometria di massa in ambito biologico e della sicurezza alimentare. Il primo capitolo tratta lo studio di aspetti metodologici ed applicativi di procedure sensoristiche per l’identificazione e la determinazione di biomarkers associati alla malattia celiaca. In tale ambito, sono stati sviluppati due immunosensori, uno a trasduzione piezoelettrica e uno a trasduzione amperometrica, per la rivelazione di anticorpi anti-transglutaminasi tissutale associati a questa malattia. L’innovazione di questi dispositivi riguarda l’immobilizzazione dell’enzima tTG nella conformazione aperta (Open-tTG), che è stato dimostrato essere quella principalmente coinvolta nella patogenesi. Sulla base dei risultati ottenuti, entrambi i sistemi sviluppati si sono dimostrati una valida alternativa ai test di screening attualmente in uso per la diagnosi della celiachia. Rimanendo sempre nel contesto della malattia celiaca, ulteriore ricerca oggetto di questa tesi di Dottorato, ha riguardato lo sviluppo di metodi affidabili per il controllo di prodotti “gluten-free”. Il secondo capitolo tratta lo sviluppo di un metodo di spettrometria di massa e di un immunosensore competitivo per la rivelazione di prolammine in alimenti “gluten-free”. E’ stato sviluppato un metodo LC-ESI-MS/MS basato su un’analisi target con modalità di acquisizione del segnale selected reaction monitoring per l’identificazione di glutine in diversi cereali potenzialmente tossici per i celiaci. Inoltre ci si è focalizzati su un immunosensore competitivo per la rivelazione di gliadina, come metodo di screening rapido di farine. Entrambi i sistemi sono stati ottimizzati impiegando miscele di farina di riso addizionata di gliadina, avenine, ordeine e secaline nel caso del sistema LC-MS/MS e con sola gliadina nel caso del sensore. Infine i sistemi analitici sono stati validati analizzando sia materie prime (farine) che alimenti (biscotti, pasta, pane, etc.). L’approccio sviluppato in spettrometria di massa apre la strada alla possibilità di sviluppare un test di screening multiplo per la valutazione della sicurezza di prodotti dichiarati “gluten-free”, mentre ulteriori studi dovranno essere svolti per ricercare condizioni di estrazione compatibili con l’immunosaggio competitivo, per ora applicabile solo all’analisi di farine estratte con etanolo. Terzo capitolo di questa tesi riguarda lo sviluppo di nuovi metodi per la rivelazione di HPV, Chlamydia e Gonorrhoeae in fluidi biologici. Si è scelto un substrato costituito da strips di carta in quanto possono costituire una valida piattaforma di rivelazione, offrendo vantaggi grazie al basso costo, alla possibilità di generare dispositivi portatili e di poter visualizzare il risultato visivamente senza la necessità di strumentazioni. La metodologia sviluppata è molto semplice, non prevede l’uso di strumentazione complessa e si basa sull’uso della isothermal rolling-circle amplification per l’amplificazione del target. Inoltre, di fondamentale importanza, è l’utilizzo di nanoparticelle colorate che, essendo state funzionalizzate con una sequenza di DNA complementare al target amplificato derivante dalla RCA, ne permettono la rivelazione a occhio nudo mediante l’uso di filtri di carta. Queste strips sono state testate su campioni reali permettendo una discriminazione tra campioni positivi e negativi in tempi rapidi (10-15 minuti), aprendo una nuova via verso nuovi test altamente competitivi con quelli attualmente sul mercato.

The role of the social environment in non-suicidal self-injury among LGBTQ youth: A mixed methods study

Non-suicidal self-injury (NSSI), such as cutting and burning, is a widespread social problem among lesbian, gay, bisexual, transgender, queer, and questioning (LGBTQ) youth. Extant research indicates that this population is more than twice as likely to engage in NSSI than heterosexual and cisgender (non-transgender) youth. Despite the scope of this social problem, it remains relatively unexamined in the literature. Research on other risk behaviors among LGBTQ youth indicates that experiencing homophobia and transphobia in key social contexts such as families, schools, and peer relationships contributes to health disparities among this group. Consequently, the aims of this study were to examine: (1) the relationship between LGBTQ youth's social environments and their NSSI behavior, and (2) whether/how specific aspects of the social environment contribute to an understanding of NSSI among LGBTQ youth. This study was conducted using an exploratory, sequential mixed methods design with two phases. The first phase of the study involved analysis of transcripts from interviews conducted with 44 LGBTQ youth recruited from a community-based organization. In this phase, five qualitative themes were identified: (1) Violence; (2) Misconceptions, Stigma, and Shame; (3) Negotiating LGBTQ Identity; (4) Invisibility and Isolation; and (5) Peer Relationships. Results from the qualitative phase were used to identify key variables and specify statistical models in the second, quantitative, phase of the study, using secondary data from a survey of 252 LGBTQ youth. The qualitative phase revealed how LGBTQ youth, themselves, described the role of the social environment in their NSSI behavior, while the quantitative phase was used to determine whether the qualitative findings could be used to predict engagement in NSSI among a larger sample of LGBTQ youth. The quantitative analyses found that certain social-environmental factors such as experiencing physical abuse at home, feeling unsafe at school, and greater openness about sexual orientation significantly predicted the likelihood of engaging in NSSI among LGBTQ youth. Furthermore, depression partially mediated the relationships between family physical abuse and NSSI and feeling unsafe at school and NSSI. The qualitative and quantitative results were compared in the interpretation phase to explore areas of convergence and incongruence. Overall, this study's findings indicate that social-environmental factors are salient to understanding NSSI among LGBTQ youth. The particular social contexts in which LGBTQ youth live significantly influence their engagement in this risk behavior. These findings can inform the development of culturally relevant NSSI interventions that address the social realities of LGBTQ youth's lives.

Examining the Alliance-Outcome Relationship: Reverse Causation, Third Variables, and Treatment Phase Artifacts

Psychotherapy research reveals consistent associations between therapeutic alliance and treatment outcomes in the youth and adult literatures. Despite these consistent findings, prospective associations are not sufficient to support the claim that the alliance is a change mechanism in psychotherapy. The current study examined the direction of effect of the alliance- outcome relationship, the contribution of early symptom change in treatment to the development of therapeutic alliance, and the potential for pretreatment interpersonal functioning characteristics to be third variables that account for the association between alliance and outcome. Participants were adolescents with depression and a history of interpersonal trauma that presented to a community mental health center for treatment. Findings demonstrated that a more positive therapeutic alliance predicted greater subsequent symptom improvement, even after removing symptom change occurring before the measurement of alliance. Results also suggested that early change only slightly contributed to alliance development. Finally, though pretreatment interpersonal functioning was related to the first session alliance, these pretreatment client characteristics were not related to later alliance or symptom change. Overall, results provided some support for therapeutic alliance as a mechanism of change in psychotherapy. Methodological and clinical issues are discussed.

Identifying the role of natural resources in knowledge-based strategies of development

Knowledge has adopted a preferential role in the explanation of development while the evidence about the effect of natural resources in countries’ performance is more controversial in the economic literature. This paper tries to demonstrate that natural resources may positively affect growth in countries with a strong natural resources specialization pattern although the magnitude of these effects depend on the type of resources and on other aspects related to the production and innovation systems. The positive trajectory described by a set of national economies mainly specialized in natural resources and low-tech industries invites us to analyze what is the combination of factors that serves as engine for a sustainable development process. With panel data for the period 1996-2008 we estimate an applied growth model where both traditional factors and other more related to innovation and absorptive capabilities are taken into account. Our empirical findings show that according to the postulates of a knowledge-based approach, a framework that combines physical and intangible factors is more suitable for the definition of development strategies in those prosperous economies dominated by natural resources and connected activities, while the internationalization process of activities and technologies become also a very relevant aspect.

The Development of an Intervention Tool to Address Challenges in the Medical Treatment of Children with Autism .

Challenges in treating children with an autism spectrum disorder (ASD) in medical settings are identified and discussed. Although research supports interventions for children with ASD including positive reinforcement, environmental modification, and visual supports and systems, limited research on the efficacy of these interventions in medical environments and with specific procedures exists. Based on the available intervention literature, this project proposes a picture schedule reinforcement system for use during blood draw procedures for ASD children with diabetes. Future efforts should include increased education for medical providers and health professionals as psychological interventions continue to inform best practices in care for children with ASD in medical settings.

Applying Positive Psychology in Early Childhood.

The purpose of this paper is to introduce a framework for applying positive psychology in elementary classrooms. The target age group is children in grades K-3 (ages 5 to 8) because this age group can benefit the most from an early introduction to strategies that promote positive development (Cowne & Hightower, 1989; White, 1996). The following sections will: (a) introduce constructs of positive psychology; (b) present developmental data on how these constructs can be applied to children ages 5 to 8 years; (c) present ideas for incorporating positive psychology practice into K-3 classrooms; (d) present strategies for incorporating positive psychology with multicultural considerations; and (e) present ideas on how to implement strategies based on positive psychology that are compatible with grade level standards and sociopolitical teaching expectations.

Invisible Suffering: Practitioner Reflections on Peacebuilding Programs with Youth Exposed to Traumatic Stressors in Intergroup Conflict

For decades, the international community has recognized that youth are some of the most vulnerable to mental and emotional distress within the intractable and cyclical nature of identity-based violent conflict. Exposure to traumatic stressors within these intergroup conflicts poses unique risks not only to the neurological and social development of youth, but also to the capacities of youth to fully participate in peacebuilding interventions. The peacebuilding field has yet to strongly consider how traumatic stress affects dynamics within programs for youth and how these programs may need to modify expectations of youth’s cognitive, social, and emotional functioning to account for the traumatic dimensions of political and social violence. Through a qualitative analysis of practitioner reflections gathered from an online survey distributed worldwide, this study explores how practitioners conceptualize and approach issues of traumatic stress in peacebuilding programs focused on youth in conflict-affected contexts. The objective is to identify the working assumptions undergirding practitioner conceptualizations and approaches to traumatic stress and gaps in trauma interventions in peacebuilding programs for youth. The implications of these findings will support efforts to enhance trauma-sensitive peacebuilding practice by revisiting and reconsidering preexisting norms.

Examining the relationship among genes, attention bias to emotion, and depression in youth

The investigation of biologically initiated pathways to psychological disorder is critical to advance our understanding of mental illness. Research has suggested that attention bias to emotion may be an intermediate trait for depression associated with biologically plausible candidate genes, such as the serotonin transporter (5-HTTLPR) and catechol-o-methyl-transferase (COMT) genes, yet there have been mixed findings in regards to the precise direction of effects. The experience of recent stressful life events (SLEs) may be an important, yet currently unstudied, moderator of the relationship between genes and attention bias as SLEs have been associated with both gene expression and attention to emotion. Additionally, although attention biases to emotion have been studied as a possible intermediate trait associated with depression, no study has examined whether attention biases within the context of measured genetic risk lead to increased risk for clinical depressive episodes over time. Therefore, this research investigated both whether SLEs moderate the link between genetic risk (5-HTTLPR and COMT) and attention bias to emotion and whether 5-HTTLPR and COMT moderated the relationship between attention biases to emotional faces and clinical depression onset prospectively across 18 months within a large community sample of youth (n= 467). Analyses revealed a differential effect of gene. Youth who were homozygous for the low expressing allele of 5-HTTLPR (S/S) and had experienced more recent SLEs within the last three months demonstrated preferential attention toward negative emotional faces (angry and sad). However, youth who were homozygous for the high expressing COMT genotype (Val/Val) and had experienced more recent SLEs showed attentional avoidance of positive facial expressions (happy). Additionally, youth who avoided negative emotion (i.e., anger) and were homozygous for the S allele of the 5-HTTLPR gene were at greater risk for prospective depressive episode onset. Increased risk for depression onset was specific to the 5-HTTLPR gene and was not found when examining moderation by COMT. These findings highlight the importance of examining risk for depression across multiple levels of analysis, such as combined genetic, environmental, and cognitive risk, and is the first study to demonstrate clear evidence of attention biases to emotion functioning as an intermediate trait predicting depression.

Examining the Complex Relationships of Early Intervention on Language and Psychosocial Development in Families Facing Multiple Risks with a Focus on Children of Hispanic Immigrants

Abundant research has shown that poverty has negative influences on young child academic and psychosocial development, and unfortunately, disparities in school readiness between low and high income children can be seen as early the first year of life. The largest federal early care and education intervention for these vulnerable children is Early Head Start (EHS). To diminish these disparate child outcomes, EHS seeks to provide community based flexible programming for infants and toddlers and their families. Given how relatively recent these programs have been offered, little is known about the nuances of how EHS impacts infant and toddler language and psychosocial development. Using a framework of Community Based Participatory Research (CBPR) this paper had 5 goals: 1) to characterize the associations between domain specific and cumulative risk and child outcomes 2) to validate and explore these risk-outcome associations separately for Children of Hispanic immigrants (COHIs), 3) to explore relationships among family characteristics, multiple environmental factors, and dosage patterns in different EHS program types, 4) to examine the relationship between EHS dosage and child outcomes, and 5) to examine how EHS compliance impacts child internalizing and externalizing behaviors and emerging language abilities. Results of the current study showed that risks were differentially related to child outcomes. Poor maternal mental health was related to child internalizing and externalizing behaviors, but not related to emerging child language skills. Although child language skills were not related to maternal mental health, they were related to economic hardship. Additionally, parent level Spanish use and heritage orientation were associated with positive child outcomes. Results also showed that these relationships differed when COHIs and children with native-born parents were examined separately. Further, unique patterns emerged for EHS program use, for example families who participated in home-based care were less likely to comply with EHS attendance requirements. These findings provide tangible suggestions for EHS stakeholders: namely, the need to develop effective programming that targets engagement for diverse families enrolled in EHS programs.

Development of multidisciplinary workshops between different subjects as an innovative teaching methodology for the acquisition of professional skills of the future architect

The implantation of the new Architecture Degree and the important normative changes in the building sector imply the need to use new teaching methodologies that enhance skills and competences in order to response to the increasing requirements demanded by society to the future architect. The aim of this paper is to present, analyze and discuss the development of multidisciplinary workshops as a new teaching methodology used in several Construction subjects of the Architecture Degree in the University of Alicante. Workshops conceived with the aim to synthesize and complement the technical knowledge acquired by the students during the Degree and to enhance the skills and competencies necessary for the professional practice. With that purpose, we decided to experiment on current subjects of the degree during this academic year, by applying the requirements defined in the future Architecture Degree in a practical way, through workshops between different subjects, superposing the technical knowledge with the resolution of constructive problems in the development of an architectural project. Developing these workshops between subjects we can dissolve the traditional boundaries between different areas of the Degree. This multidisciplinary workshop methodology allows the use of all the global knowledge acquired by students during their studies and at the same time, it enhances students’ ability to communicate and discuss their ideas and solutions in public. It also increases their capacity of self-criticism, and it foments their ability to undertake learning strategies and research in an autonomous way. The used methodology is based on the development of a practical work common to several subjects of different knowledge areas within the "Technology Block" of the future Architecture Degree. Thus, students work approaching the problem in a global way discussing simultaneously with teachers from different areas. By using these new workshops we stimulate an interactive class versus a traditional lecture. Work is evaluated continuously, valuing the participative pupil´s attitude, working in groups in class time, reaching weekly objectives and stimulating the individual responsibility and positive interdependence of the pupil inside the working group. The exercises are designed to improve students’ ability to transmit their ideas and solutions in public, knowing how to discuss and defend their technical resolutions to peers and teachers (Peer Reviewing), their capacity for self-criticism and their capacity to undertake strategies and autonomous learning processes at the same time they develop a personal research into new technologies, systems and materials. Students have shown their majority preference for this teaching methodology by the multidisciplinary workshops offered in the last years, with very satisfactory academic results. In conclusion, it can be verified nowadays the viability of the introduction of new contents and new teaching methodologies necessary for the acquisition of the skills in the future Architecture Degree, through workshops between several subjects that have had a great acceptance in students and positive contrasted academic results.

Community Size in Youth Sport Settings: Examining Developmental Assets and Sport Withdrawal

Despite the popularity of youth sport programs, little research has examined the psychosocial benefits assumed to stem from involvement. Some studies suggest birthplace influences the development of elite athletes, but little work has examined other influences of community contexts. The purpose of this study was to examine relationships between young athletes’ community size, developmental assets, and sport involvement. Current and recently withdrawn competitive swimmers (N = 181) completed the Developmental Assets Profile (Search Institute, 2004). Athletes from smaller cities had significantly higher developmental asset scores for support, commitment to learning, and boundaries/expectations. Further, community size was a significant predictor of withdrawal. Findings suggest community context should be given additional attention in youth sport literature.

Developmental Profiles of Successful High School Coaches

The purpose of this exploratory study was to compare the developmental profiles of successful high-school sport coaches, and to determine if elements of a coach’s developmental profile were associated with coaching success. Sixteen high-school coaches in the United States – nine who coach basketball and seven cross-country running – participated in structured retrospective quantitative interviews. All coaches had accumulated extensive experience as an athlete (M = 19.6 seasons; 2,428.8 hours) and were better than average athletes in relation to their peers. Positive significant relationships were found between time (seasons and hours) spent as an athlete in the sport that the participants now coach and five measures of coaching success. The results are discussed in relation to the ongoing dialogue about coach development, coaching effectiveness, and coach education.

Transfer of Pattern Recall Skills May Contribute to the Development of Sport Expertise

Superior recall of domain-specific patterns is well established as a defining attribute of expert performers. Recent studies on the developmental histories of expert team ball sport players (e.g. Baker, Côté, & Abernethy, 2003a) also suggest that experts characteristically receive exposure to a wide range of sports in their developing years and that this related sports experience may reduce the amount of sport-specific training needed to become an expert. This study examined whether the facilitation of expertise associated with other sport experience might arise from positive transfer of pattern recall skills from one sport to another. Expert netball, basketball and field hockey players and experienced non-experts performed a recall task for patterns of play derived from each of these sports. Experts from sports different to those shown in the presented pattern consistently outperformed non-experts in their recall of defensive player positions, suggesting some selective transfer of pattern recall skills may indeed be possible