Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Effects of Morphogen and Scaffold Porogen on the Differentiation of Dental Pulp Stem Cells

Introduction: Dental pulp tissue engineering is an emerging field that can potentially have a major impact on oral health. However, the source of morphogens required for stem cell differentiation into odontoblasts and the scaffold characteristics that are more conducive to odontoblastic differentiation are still unclear. This study investigated the effect of dentin and scaffold porogen on the differentiation of human dental pulp stem cells (DPSCs) into odontoblasts. Methods: Poly-L-lactic acid (PLLA) scaffolds were prepared in pulp chambers of extracted human third molars using salt crystals or gelatin spheres as porogen. DPSCs seeded in tooth slice/scaffolds or control scaffolds (without tooth slice) were either cultured in vitro or implanted subcutaneously in immunodefficient mice. Results: DPSCs seeded in tooth slice/scaffolds but not in control scaffolds expressed putative odontoblastic markers (DMP-1, DSPP, and MEPE) in vitro and in vivo. DPSCs seeded in tooth/slice scaffolds presented lower proliferation rates than in control scaffolds between 7 and 21 days (p \u3c 0.05). DPSCs seeded in tooth slice/scaffolds and transplanted into mice generated a tissue with morphological characteristics similar to those of human dental pulps. Scaffolds generated with gelatin or salt porogen resulted in similar DPSC proliferation. The porogen type had a relatively modest impact on the expression of the markers of odontoblastic differentiation. Conclusions: Collectively, this work shows that dentin-related morphogens are important for the differentiation of DPSC into odontoblasts and for the engineering of dental pulp-like tissues and suggest that environmental cues influence DPSC behavior and differentiation potential

Tratamento do transtorno de pânico com terapia psicodramática de grupo Treatment for panic disorder with psychodramatic group therapy

INTRODUÇÃO/OBJETIVOS: Há poucos trabalhos publicados sobre psicoterapia grupal para pacientes com transtorno de pânico (TP); além disso, esses estudos geralmente são restritos a abordagens cognitivo-comportamentais. O objetivo deste trabalho é relatar uma experiência de atendimento psicoterápico psicodramático grupal para portadores de TP, iniciada em 1996 na Faculdade de Medicina de Botucatu/Unesp, e discutir aspectos psicodinâmicos desse transtorno. MÉTODOS: Atende-se uma média de oito a dez pacientes em sessões mensais de duas horas de duração, nas quais se utilizam técnicas psicodramáticas. O uso associado de psicofármacos é a regra. A temática é aberta, centrando-se tanto em aspectos próprios do TP (sintomas mais comuns; peregrinação em serviços médicos até o diagnóstico; preocupações hipocondríacas; limitações e dependência; reações dos familiares; estratégias de exposição e enfrentamento; efeitos e reações dos medicamentos) quanto em problemas individuais específicos. RESULTADOS: A abordagem psicodramática favorece a identificação e a elaboração de aspectos psicodinâmicos. Entre estes, destacaram-se os sentimentos de desamparo e seus desdobramentos: insegurança; fragilidade e medo; raiva e culpa; sentimentos de desamor e rejeição; vergonha e inferioridade; isolamento e dificuldade de pedir ajuda; dificuldade de identificar e de expressar sentimentos - manifestados somaticamente -; e dificuldade de assumir o papel de "cuidador" e outras responsabilidades. CONCLUSÕES: Aspectos valiosos para o tratamento do TP, como apoio mútuo, companheirismo, confiança, modelo e estímulo, são favorecidos pelo contexto grupal homogêneo. O compartilhar de experiências e sofrimentos comuns propiciou rápida coesão e suporte grupal, melhora da capacidade de expressar sentimentos e da auto-estima e o aprimoramento de papéis sociais. Além da melhora dos sintomas, possibilitou-se a melhor compreensão e elaboração destes, que passam a ser integrados significativamente no contexto existencial.<br>INTRODUCTION: There are few published articles on group psychotherapy for patients with panic disorder (PD) and usually they are restricted to the cognitive-behavioral approach. OBJECTIVE: To describe an experience of group psychodramatic therapy for PD patients, started in 1996 at Botucatu Medical School - UNESP, and discuss psychodynamic aspects of the panic disorder. METHODS: Two-hour monthly psychodramatic sessions, with an average of 8 to10 patients were carried out. The patients were usually receiving concomitantly psychopharmacological treatment. The subjects to be discussed were freely chosen, ranging from issues directly related to PD (most common symptoms, hurdles to get to a diagnosis, hypochondriac concerns, limitations and dependency, family reactions, strategies of exposure and coping, and effects of drug treatment) to individual conflicts and problems. RESULTS: The psychodramatic approach favors the identification and elaboration of psychodynamic aspects. The most common feelings observed were helplessness and other related ones, such as: insecurity; vulnerability and fear; anger and guilt; isolation and difficulty in identifying and expressing emotions (which manifested physically); and difficulty in assuming the role of caretaker and other responsibilities. CONCLUSIONS: Valuable aspects for the treatment of PD, such as mutual support, partnership, confidence, modeling and encouragement, are favored in a group setting, where sharing experiences and sufferings helped to improve the participants' self-esteem, their ability to express emotions and to play different social roles. Besides the clinical improvement, the symptoms could be better understood and elaborated, and meaningfully integrated into their life context