Silvana Aparecida Alves Corrêa de Noronha

Toll like receptors gene expression of human keratinocytes cultured of severe burn injury

PURPOSE To evaluate the expression profile of genes related to Toll Like Receptors (TLR) pathways of human Primary Epidermal keratinocytes of patients with severe burns. METHODS After obtaining viable fragments of skin with and without burning, culture hKEP was initiated by the enzymatic method using Dispase (Sigma-Aldrich). These cells were treated with Trizol(r) (Life Technologies) for extraction of total RNA. This was quantified and analyzed for purity for obtaining cDNA for the analysis of gene expression using specific TLR pathways PCR Arrays plates (SA Biosciences). RESULTS After the analysis of gene expression we found that 21% of these genes were differentially expressed, of which 100% were repressed or hyporegulated. Among these, the following genes (fold decrease): HSPA1A (-58), HRAS (-36), MAP2K3 (-23), TOLLIP (-23), RELA (-18), FOS (-16), and TLR1 (-6.0). CONCLUSIONS This study contributes to the understanding of the molecular mechanisms related to TLR pathways and underlying wound infection caused by the burn. Furthermore, it may provide new strategies to restore normal expression of these genes and thereby change the healing process and improve clinical outcome.

Innate and adaptive immunity gene expression of human keratinocytes cultured of severe burn injury

PURPOSE Evaluate the expression profile of genes related to Innate and Adaptive Immune System (IAIS) of human Primary Epidermal keratinocytes (hPEKP) of patients with severe burns. METHODS After obtaining viable fragments of skin with and without burning, culture hKEP was initiated by the enzymatic method using Dispase (Sigma-Aldrich). These cells were treated with Trizol(r) (Life Technologies) for extraction of total RNA. This was quantified and analyzed for purity for obtaining cDNA for the analysis of gene expression using specific IAIS PCR Arrays plates (SA Biosciences). RESULTS After the analysis of gene expression we found that 63% of these genes were differentially expressed, of which 77% were repressed and 23% were hyper-regulated. Among these, the following genes (fold increase or decrease): IL8 (41), IL6 (32), TNF (-92), HLA-E (-86), LYS (-74), CCR6 (- 73), CD86 (-41) and HLA-A (-35). CONCLUSIONS This study contributes to the understanding of the molecular mechanisms underlying wound infection caused by the burn. Furthermore, it may provide new strategies to restore normal expression of these genes and thereby change the healing process and improve clinical outcome.

Abstract 72: keratinocytes gene expression of innate and adaptative proinflammatory cytokines and members of the toll-like receptor pathways in severe burned.

Sarday, M arch 8, 2014 inguinal nodes 3 hours following injection, tortuous collateral lymphatic channels, and dermal backflow consistent with lymphedema. Arrows indicate inguinal nodes, black arrowheads show tortuous lymphatic channels and dermal backflow, and white arrowheads mark the feet where the radiolabelled tracer was injected. 72 Keratinocytes Gene expression of innate and adaptative Proinflammatory cytokines and Members of the toll-like receptor Pathways in Severe Burned

SOCIAL INCLUSION AND SOLIDARITY IN STRICT POST-GRADUATION: EXPERIENCING THE LABORATORY "SCIENTIFIC WAKE FOR THE FUTURE" (PRIMARY AND SECONDARY EDUCATION)

OBJECTIVE To stimulate and awakening the interest of students of high school or elementary public schools in research and science through scientific initiation stages in the Postgraduate Program in Translational Surgery. To stimulate and awakening the interest of students of high school or elementary public schools in research and science through scientific initiation stages in the Postgraduate Program in Translational Surgery. METHOD The target audience for the development of scientific activities were students enrolled in mid-level course (second year initially) and have approval of their participation in this project by the school and by legal guardians. The inclusion criteria were: physical proximity to the higher education institution, signing the consent form by the legal responsible for the students, and for the board of the school unit and the researcher. Initially, students performed diagnostic evaluation about the prior knowledge of biology, science and scientific research. From there, the classes were prepared based on the result of this test, then started the activities of Junior Scientific Initiation in basic education. RESULTS The school chosen for this initial phase of the pilot project was the State School Rui Bloem which has 13 classrooms for the second year of medium education in a total of 390 students. Of these, 160 (41%) were interested but only 16 (10%) were eligible to start the pilot project in Translational Surgery Laboratory of Unifesp. These students showed average yield of 50% in diagnostic test and should start the next training in cell and molecular biology laboratory and also to attend scientific meetings. CONCLUSION In the initial phase of the project, was observed the great student interest in scientific career, but at the same time, a great need for improvement. The choice of public school was for access to university and proximity. In addition, these students have more shortcomings and deficiencies. But this does not mean that the fascination for scientific career cannot turn them into great researchers thus contributing to the economic, social and intellectual growth of our country through scientific research. OBJETIVO Estimular e despertar o interesse dos alunos do ensino médio ou fundamental de escolas públicas na pesquisa e na ciência por meio de estágios de Iniciação Científica Junior no Programa de Pós-Graduação em Cirurgia Translacional da Unifesp. MÉTODO O público alvo para o desenvolvimento das atividades científicas foram alunos regularmente matriculados em curso de nível médio (segundos anos inicialmente) e que tivessem aprovação de sua participação pela direção da escola e responsáveis legais. Quanto aos critérios de inclusão: proximidade física com a instituição de ensino superior, assinatura do termo de consentimento pelos responsáveis dos alunos, pela diretoria da unidade escolar e pelo pesquisador. Inicialmente, os alunos realizaram avaliação diagnóstica acerca dos conhecimentos prévios de biologia, ciências e pesquisa científica. A partir daí as aulas eram elaboradas com base no resultado deste teste, para então iniciar as atividades de Iniciação Científica Junior. RESULTADOS A escola escolhida para esta fase inicial do projeto piloto foi a Escola Estadual Rui Bloem que possui 13 salas de aula para o segundo ano do ensino médio com total de 390 alunos. Destes, 160 (41%) apresentaram-se interessados; porém, somente 16 (10%) foram elegíveis para iniciar o projeto piloto no Laboratório de Cirurgia Translacional da Unifesp. Estes alunos apresentaram rendimento médio de 50% na prova diagnóstica e deverão iniciar os próximos treinamentos no Laboratório de Biologia Celular e Molecular e também a frequentar as reuniões científicas. CONCLUSÃO Nesta fase inicial houve interesse dos alunos do ensino médio e o projeto piloto apresentado estimulou e despertou interesse dos alunos de escola pública na pesquisa e na ciência. A escolha de uma escola pública foi para o acesso à universidade e proximidade. Além disso, esses alunos têm mais carências e deficiências. Mas isso não significa que o fascínio pela carreira científica não pode transformá-los em grandes pesquisadores, contribuindo assim para o crescimento econômico, social e intelectual do nosso país através da pesquisa científica.