FM1-43 dye ultrastructural localization in and release from frog motor nerve terminals.

Previous work has shown that the fluorescent styryl dye FM1-43 stains nerve terminals in an activity-dependent fashion. This dye appears to label the membranes of recycled synaptic vesicles by being trapped during endocytosis. Stained terminals can subsequently be destained by repeating nerve stimulation in the absence of dye; the destaining evidently reflects escape of dye into the bathing medium from membranes of exocytosing synaptic vesicles. In the present study we tested two key aspects of this interpretation of FM1-43 behavior, namely: (i) that the dye is localized in synaptic vesicles, and (ii) that it is actually released into the bathing medium during destaining. To accomplish this, we first photolyzed the internalized dye in the presence of diaminobenzidine. This created an electron-dense reaction product that could be visualized in the electron microscope. Reaction product was confined to synaptic vesicles, as predicted. Second, using spectrofluorometry, we quantified the release of dye liberated into the medium from tubocurarine-treated nerve-muscle preparations. Nerve stimulation increased the amount of FM1-43 released, and we estimate that normally a stained synaptic vesicle contains a few hundred molecules of the dye. The key to the successful detection of released FM1-43 was to add the micelle-forming detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), which increased FM1-43 quantum yield by more than two orders of magnitude.

The secreted Ipa complex of Shigella flexneri promotes entry into mammalian cells.

The bacterial pathogen Shigella flexneri causes bacillary dysentery in humans by invading coloncytes. Upon contact with epithelial cells, S. flexneri elicits localized plasma membrane projections sustained by long actin filaments which engulf the microorganism. The products necessary for Shigella entry include three secretory proteins: IpaB, IpaC, and IpaD. Extracellular IpaB and IpaC associate in a soluble complex, the Ipa complex. We have immunopurified this Ipa complex on latex beads and found that they were efficiently internalized into HeLa cells. Like S. flexneri entry, uptake of the beads bearing the Ipa complex was associated with membrane projections and polymerization of actin at the site of cell-bead interaction and was dependent on small Rho GTPases. These results indicate that a secreted factor can promote S. flexneri entry into epithelial cells.

Enterococcus faecalis pheromone binding protein, PrgZ, recruits a chromosomal oligopeptide permease system to import sex pheromone cCF10 for induction of conjugation.

Conjugative transfer of the plasmid pCF10 by Enterococcus faecalis donor cells occurs in response to a peptide sex pheromone, cCF10, secreted by recipients. The plasmid-encoded cCF10 binding protein, PrgZ, is similar in sequence to binding proteins (OppAs) encoded by oligopeptide permease (opp) operons. Mutation of prgZ decreased the sensitivity of donor cells to pheromone, whereas inactivation of the chromosomal E. faecalis opp operon abolished response at physiological concentrations of pheromone. Affinity chromatography experiments demonstrated the interaction of the pheromone with several putative intracellular regulatory molecules, including an RNA molecule required for positive regulation of conjugation functions. These data suggest that processing of the pheromone signal involves recruitment of a chromosomal Opp system by PrgZ and that signaling occurs by direct interaction of internalized pheromone with intracellular effectors.

Agonist-induced desensitization of dopamine D1 receptor-stimulated adenylyl cyclase activity is temporally and biochemically separated from D1 receptor internalization.

The regulation of the dopamine D1 receptor was investigated by using c-myc epitope-tagged D1 receptors expressed in Sf9 (fall armyworm ovary) cells. Treatment of D1 receptors with 10 microM dopamine for 15 min led to a loss of the dopamine-detected high-affinity state of the receptor accompanying a 40% reduction in the ability of the receptor to mediate maximal dopamine stimulation of adenylyl cyclase activity. After 60 min of agonist exposure, 45 min after the occurrence of desensitization, 28% of the cell surface receptors were internalized into an intracellular light vesicular membrane fraction as determined by radioligand binding and supported by photoaffinity labeling, immunocytochemical staining, and immunoblot analysis. Pretreatment of cells with concanavalin A or sucrose completely blocked agonist-induced D1 receptor internalization without preventing agonist-induced desensitization, indicating a biochemical separation of these processes. Collectively, these findings indicate that the desensitization of D1 receptor-coupled adenylyl cyclase activity and D1 receptor internalization are temporarily and biochemically distinct mechanisms regulating D1 receptor function following agonist activation.

Folate receptors targeted to clathrin-coated pits cannot regulate vitamin uptake.

Potocytosis is an endocytic process that is specialized for the internalization of small molecules. Recent studies on the uptake of 5-methyltetrahydrofolate by the folate receptor have suggested that the glycosyl-phosphatidylinositol anchor on this protein causes it to cluster and be internalized by caveolae instead of coated pits. To test this hypothesis directly, we have constructed a chimeric folate receptor that has the glycosyl-phosphatidylinositol anchor replaced with the transmembrane domain and cytoplasmic tail of the low density lipoprotein receptor. The cells with wild-type receptors delivered 5-methyltetrahydrofolate to the cytoplasm more rapidly than did cells expressing the chimeric receptor. This suggests that efficient delivery to the cytoplasm depends on caveolae. In sharp contrast to cells with wild-type folate receptors, cells internalizing folate by clathrin-coated pits were unable to decrease vitamin uptake when they were either folate replete or confluent.

Significados da reinserção escolar de crianças após queimaduras: um estudo etnográfico

As queimaduras na infância compreendem importante causa de atendimento hospitalar e internação e podem desencadear sequelas físicas e emocionais. Dependendo da gravidade e complexidade das lesões, a hospitalização da criança é prolongada para realização de tratamento adequado e ela é afastada tanto do convívio familiar como do social. O número de crianças em idade escolar que sofrem queimaduras, e consequentemente interrompem as atividades escolares, por períodos curtos ou até as abandonam, é significativo, tornando digno de preocupação o processo de reinserção escolar. Este estudo teve como objetivo interpretar os significados do processo de reinserção escolar de crianças sobreviventes de queimaduras. Realizou-se uma pesquisa com abordagem metodológica qualitativa, fundamentada no referencial teórico da Antropologia Interpretativa e no método etnográfico. Após a aprovação do estudo pelos Comitês de Ética em Pesquisa com Seres Humanos das instituições envolvidas, convidaram-se crianças sobreviventes de queimaduras em seguimento em um hospital-escola do interior paulista. Coletaram-se dados no hospital, no domicílio e na escola das crianças, de janeiro de 2012 a dezembro de 2013, por meio de entrevistas em profundidade audiogravadas e observação participante, complementadas pelo diário de campo. Participaram da pesquisa 14 crianças e os atores sociais envolvidos neste processo, como familiares, profissionais de saúde, professores, vizinha e amigo, totalizando 57 participantes. A coleta e análise dos dados ocorreram concomitantemente, e esta última seguiu os pressupostos da análise temática indutiva. Identificaram-se códigos, os quais, posteriormente, embasaram a construção das duas unidades de sentidos: \"Fatos e fatores prévios ao retorno escolar\" e \"A volta à escola\". A partir destas, identificou-se o núcleo temático, \"O olho puxa essas coisas. A gente olha mesmo\": enxergando o outro como diferente, o qual apresenta a explicação compreensiva do processo de reinserção escolar, fornecendo os significados da experiência. Os significados foram explicados por meio dos conceitos antropológicos de estigma, identidade e corporeidade. O culto ao corpo presente no contexto cultural brasileiro foi fator intensificador do estigma sofrido pelas crianças. Identificaram-se fatores facilitadores do processo, como: dar continuidade às atividades escolares e manter contato com professores e colegas durante a hospitalização; preparar a escola para receber a criança; abordar sobre queimadura na escola para evitar curiosidade; encarar a presença da discriminação e trabalhar as diferenças. Estas estratégias fazem com que todos os envolvidos se sintam ao menos mais confortáveis durante esse difícil processo que merece atenção e envolvimento dos familiares, profissionais de saúde e equipe escolar. A partir dos resultados desta pesquisa, esperamos empoderar as crianças e seus familiares acerca da melhor maneira para lidar com as queimaduras durante a reinserção escolar e contribuir para o desenvolvimento de ações e estratégias baseadas na cultura para que este processo ocorra da forma menos traumática possível

Saúde mental e os novos paradigmas de cidadania e inclusão social na sociedade contemporânea

Este estudo tem por objetivo analisar as categorias de cidadania e inclusão social na política de desinstitucionalização nos sujeitos em sofrimento psíquico diante do processo de individualização na teoria social contemporânea. Assume como hipótese que a saída do hospital psiquiátrico por si só não garante a inclusão social e nem o livre exercício da cidadania. Considerado o objetivo desta pesquisa, optou-se por fazer uma pesquisa bibliográfica como procedimento metodológico. O material de estudo foi dividido em três conjuntos: (1) 56 artigos científicos, visando a compreender a visão da academia; (2) um conjunto de legislação, composto de 10 leis que implementaram a política de desinstitucionalização no Brasil e a reforma dos serviços de saúde psiquiátrica, visando a compreender as ações do Estado; (3) quatro Relatórios Finais das quatro Conferências Nacionais de Saúde Mental, para também compreender a participação da sociedade civil. Para a análise do material, utilizou-se uma combinação de duas técnicas complementares: leitura bibliográfica com a análise de conteúdo. Dentre os vários processos que caracterizam a sociedade contemporânea, optou-se por analisar a individualização que impacta nas formas de exercício da cidadania e na inclusão social. Na análise dos resultados da categoria de cidadania foram identificadas associações em relação à interdição civil, liberdade, moradia, saúde, trabalho, educação e participação política. Relacionadas à categoria de inclusão social foram identificadas as referências à família, estigma, laços sociais, autonomia, contratualidade e trabalho. Os resultados obtidos indicam que o campo da saúde mental não está em completa consonância com as transformações da sociedade contemporânea, o que provoca um descolamento da realidade social da própria politica de desinstitucionalização e, portanto, maior dificuldade para a efetiva inclusão social e o exercício da cidadania desses indivíduos.

Instrumento de qualidade de vida relacionada à saúde para crianças e adolescentes brasileiros: validação do módulo específico HIV-DISABKIDS

Crianças e adolescentes com HIV/AIDS necessitam de uma abordagem compreensiva e singular, pois particularidades como a revelação do diagnóstico, o estigma gerado pela doença, as consultas médicas frequentes, o uso diário de medicações e os seus efeitos colaterais são aspectos que necessitam ser avaliados, quando se trata da qualidade de vida relacionada à saúde-QVRS. A inclusão da avaliação da qualidade de vida relacionada à saúde dessa população, com instrumentos viáveis e padronizados, pode identificar parâmetros para um cuidado integral e de acordo com as necessidades individuais. A presente pesquisa teve como objetivo validar o instrumento do módulo específico do DISABKIDS® de mensuração da Qualidade de Vida Relacionada à Saúde de Crianças e Adolescentes brasileiros com HIV, denominado \"Viver com HIV\". Trata-se de um estudo metodológico, do tipo validação de instrumento para o qual utilizamos a adaptação metodológica descrita pelo projeto europeu DISABKIDS®. Participaram da validação de face e conteúdo 15 especialistas com formação diversificada na área da saúde, os quais foram convidados a analisar a importância dos itens selecionados e a adequação para a faixa etária e população, classificando-os de acordo com a relevância, clareza e aplicabilidade de seu conteúdo para a população em estudo. Na validação semântica, participaram 16 crianças e adolescentes e seus respectivos pais ou cuidadores que seguiam em tratamento em uma Unidade Especializada em Tratamento de Doenças Infecciosas de um hospital universitário do Estado de São Paulo. Esta etapa consistiu em analisar, em cada item do instrumento, os termos empregados, por meio de entrevistas com a população para a qual o instrumento se destinou. O instrumento foi validado e aceito tanto na validação de face e conteúdo quanto na semântica. A avaliação da qualidade de vida de crianças e adolescentes com HIV é primordial para melhor condução do tratamento, para isto, faz-se necessária à utilização de um instrumento adequado para a faixa etária e específico para a condição HIV. A avaliação da qualidade de vida relacionada à saúde é primordial para o manejo de crianças e adolescentes com HIV, e o instrumento \"Viver com HIV\" poderá se constituir em uma ferramenta válida e confiável para mensuração da qualidade de vida relacionada à saúde de crianças e adolescentes que vivem com HIV, podendo melhorar a qualidade da assistência em saúde e o acompanhamento dessa população

Desenvolvimento floral e do óvulo e aspectos da reprodução em Aechmea sp. e Vriesea sp. (Bromeliaceae)

A utilização de Bromélias tem sido crescente no mercado de plantas onamentais, por outro lado, muitas espécies encontram-se ameaçadas, grande parte pelos impactos humanos no ambiente. Aechmea correia-araujoi E. Pereira & Moutinho, Aechmea gamossepala Wittm, Vriesea ensiformis (Vell.) Beer e Vriesea saundersii (Carrière) E. Morren ex Mez, espécies nativas da Mata Atlântica brasileira, têm sido alvo de extrativismo. Informações básicas sobre a espécie são essenciais para subsidiar a condução de programas de conservação e melhoramento genético, que aliados a ferramentas biotecnológicas permitem a incorporação de estratégias inovadoras aos métodos de melhoramento. Neste sentido, o objetivo do presente trabalho foi descrever essas espécies, quanto à micromorfologia floral, aspectos reprodutivos envolvidos no processo de polinização, desenvolvimento floral e deesenvolvimento gametofítico, como mecanismo de preservação e produção comercial. A caracterização morfológica e anatômica das flores das espécies de Aechmea e Vriesea contribuiu para a compreensão do processo reprodutivo. As espécies apresentam grãos de pólen com alta capacidade reprodutiva, viabilidade polínica superior a 93%, germinação in vitro maior que 80% e o estigma apresenta-se receptivo da antese ao final do dia. A ontogênese floral de A. correia-araujoi é centrípeta, os primórdios desenvolvem-se na ordem, sépala, pétala, androceu e gineceu. O apêndice petalar é formado na fase final do desenvolvimento. O primórdio de óvulo tem origem placentária e caráter trizonal, o óvulo é anátropo, bitegumentado e crassinucelado. O meristema floral de A. gamosepala se desenvolve de forma centrípeta, de forma unidirecional reversa. O estigma diferencia-se na fase inicial do desenvolvimento e os apêndices petalares, na fase final. O óvulo é anátropo, crassinucelado, bitegumentado, tétrade linear, megásporo calazal funcional, desenvolvimento tipo monospórico e Polygonum. As anteras são bitecas, tetraesporangiadas, com tapete secretor. Botões florais de 8,7 - 13,0 mm são indicados no estudo de embriogênese a partir de micrósporo. As alterações celulares e o padrão de distribuição de pectinas e AGPs foram caracterizadas por análise citoquímica com azul de toluidina, KI e DAPI e imunocitoquímica por imunofluorescência com os anticorpos para RNA, pectinas esterificadas (JIM7), não esterificadas (JIM5) e AGPs (LM2, LM6, MAC207, JIM13, JIM14) e analisadas por microscopia de fluorescência. Foram caracterizados padrões de distribuição espaço-temporal de pectinas e AGP que podem ser utilizados como marcadores de desenvolvimento gametofítico masculino. As observações feitas nesse trabalho fornecem dados sobre aspectos reprodutivos das espécies que podem ser utilizados em programas de melhoramento genético, conservação e desenvolvimento de haploides

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 Impact of Child Psychologists' Stigmatization of Parents on Treatment Outcomes Through a Developmental Lens

The purpose of this paper is to examine how child psychologists' specialized training inhuman development may make them more prone to stigmatize the parents of their young clients. The stigmatization of parents may lead to fewer parents seeking treatment for their children and to poorer treatment outcomes for those who work with a child psychologist. The process of stigmatization is summarized to provide context for the method through which parents receive stigma. A commonly used theory of child development, Erik Erikson's stages of ego development, is outlined to provide background on how child psychologists may interpret and evaluate a child'sdevelopment. Child psychologists' may identify parenting practices that seem to hinder or stunt children's emotional development, which would make the psychologist more aptto stigmatize and isolate parents from the treatment process. To demonstrate the unique ways in which a child psychologist may stigmatize parents of children at different developmental stages two case studies are included. Finally, a theoretical model of treatment is described that may be more inclusive, and less stigmatizing of parents. This model outlines how the parents' concerns about and observations of their children should be validated and reflected in the treatment process. This treatment modality would allow for child psychologists to more actively involve parents in treatment and provide more education and support around their children's unique emotional development needs. Through this treatment model and child psychologists' awareness of and attempts to reduce the stigmatization of parents, treatment outcomes for young clients may improve.

An Acceptance and Commitment Approach to Issues of Behavioral Health Care in Soldiers

For over a decade, the U.S. military has been engaged in two distinct, yet equally deadly conflicts: Operation Iraqi Freedom (OIF) and Operation Enduring Freedom (OEF). There are many physical and psychological effects of war necessitating the activation and interventions of a myriad of behavioral health professionals. The purpose of the paper was to understand how and if contemporary military culture may work to support or hinder application of an Acceptance and Commitment Therapy (ACT) approach to issues of psychological health among Soldiers. While the empirical research on efficacy with Soldiers is limited, a review of military culture revealed the promotion of rigid rule following, although effective in combat, influences the emotional control agenda and stigma while in garrison. However, empirical research demonstrating the clinical benefits and flexibility of ACT is rapidly emerging with civilian and Veteran populations. Suggested as a prevention technique utilized early in Soldier's training to increase psychological flexibility, ACT appears to demonstrate much promise in ameliorating the psychological consequences of war.

Establishing Culturally Sensitive Therapeutic Communities in Ghana for Individuals with Severe and Persistent Mental Illnesses

This paper makes a proposal for the establishment of therapeutic communities for people with severe and persistent mental illnesses in Ghana. It discusses the history and features of therapeutic communities, as well as the elements that make it compatible with the agenda of the new 2012 Ghana mental health bill. This paper also discusses the present state of mental health care in this West African country and how the establishment of therapeutic communities will promote recovery of people with severe and persistent mental illness, and change the perception of chronic mental illness in Ghana. A discussion of potential modifications of the therapeutic community is offered as well as justifications for maintaining other structural aspects should this establishment materialize in Ghana. The costs of setting up therapeutic communities in this third world country are addressed with the offered conclusion that costs far outweigh the benefits. Finally, given the endeavor of the proposed therapeutic communities to assist in deinstitutionalization of care, cautions are made in this paper to ensure that the trends experienced in the United States with deinstitutionalization are not replicated in Ghana. A proposal is made in the conclusion for Ghana to move past therapeutic communities when developmentally able- to community mental health centers which were in part established to account for some of the fallouts of deinstitutionalization by providing a comprehensive and extensive range of services for people with severe and persistent mental illness.

Assessment and Promotion of Partner Support: A Prevention-Focused Intervention for Perinatal Depression in Primary Care

Perinatal depression significantly impacts the mother, her partner, the unborn fetus, and the infant/child after delivery. A review of the literature supports the need for preventive intervention methods as research has shown that even with successful treatment, disruptions in attachment, temperament, and cognitive development often remain. Primary care settings are ideal targets for prevention given that they can reach a number of people at low-cost without the stigma associated with seeking help in a mental health facility. This paper purposes a preventive intervention method for perinatal depression that can be implemented in primary care settings in both Western and non-Western countries. The intervention targets two of the primary risk factors for perinatal depression; partner support and relationship quality. The intervention is structured around key target periods in gestational development and during the early weeks after delivery. Suggestions for each target visit are based on prior research that has demonstrated how psychoeducation about the transition to parenthood, as well as increased communication, can positively affect partner support and relationship quality. The ultimate goal of the intervention is not only to prevent perinatal depression but also to improve the mental health and wellbeing of the entire family system.

Psychosocial Distress Screening in a Community Cancer Center: An Exploratory Study of Distress Screening Across the Cancer Treatment Continuum

Purpose: The primary goal of this exploratory study is to demonstrate that distress screening across the course of cancer treatment is possible and provides valuable information about patient needs over time. Distress screening is aligned with guidelines from national accrediting organizations and may lead to improved health-related quality of life, satisfaction with medical care, and possibly survival.Methods: Medical, surgical, and radiation oncology patients completed a screening instrument before their appointments during a six-month period. Patients indicated their level of distress on four domains (practical, emotional, health and social concerns). De-identified data was collected, aggregated and descriptive statistics were analyzed.Results: Approximately 3000 screens were collected and 1500 cancer patients were screened. Of patients who indicated distress, 54% demonstrated a distress level of five or greater. Distress level eight was the most frequent level of distress indicated. The Cancer Dietitian was the most commonly requested healthcare team provider. The Health Concern domain was most frequently endorsed.Conclusion: NCCN, IOM and COC guidelines recommend distress screening in all cancer treatment centers, however implementation has proven difficult. This study adds to the literature about distress in cancer patients, demonstrates the feasibility of repeated distress screening and provides a model program demonstrating the implementation of repeated distress screening at a community cancer center. Findings highlight the importance of supportive oncology services due to the prevalence of high levels of distress. Findings demonstrate the importance of the Cancer Dietitian in supportive cancer care. Additionally, the research reveals a potential perceived stigma in seeking psychosocial oncology services.