Aplicabilidade da auriculoterapia para reduzir estresse e como estratégia de coping em profissionais de enfermagem

OBJETIVOS: trata-se de ensaio clínico randomizado com o objetivo de avaliar a auriculoterapia, para diminuição de níveis de estresse, em 75 profissionais de enfermagem de um hospital e analisar os principais domínios de coping que se modificaram após o tratamento. METODOLOGIA: os sujeitos foram divididos em grupos (controle, agulhas, sementes), receberam 8 sessões nos pontos Shenmen, rim e tronco cerebral. O grupo controle não recebeu nenhuma intervenção. RESULTADOS: constataram-se diferenças significativas pela ANOVA, para níveis de estresse entre o grupo agulha/controle na terceira e quarta avaliações, segundo escores de estresse da Lista de Sintoma de Stress de Vasconcelos. Para o Inventário de Folkman e Lazarus, obteve-se diferença estatística para o domínio afastamento entre os grupos agulha/controle, após tratamento; e, na análise dentro do mesmo grupo, obtiveram-se diferenças significativas para o domínio confronto, na quarta avaliação entre grupos agulha/controle e para suporte social, na terceira avaliação para os grupos agulha/semente. CONCLUSÕES: a auriculoterapia com agulhas diminuiu os níveis de estresse significativamente e houve diminuição da utilização dos domínios afastamento, confronto e suporte social para os grupos de intervenção, após o tratamento, sugerindo que a auriculoterapia tanto com agulhas quanto com sementes pode produzir impacto positivo para melhorar a estratégia de coping na equipe de enfermagem. Mais estudos, porém, fazem-se necessários para que se possa conhecer o alcance da técnica.

RNA extraction from ten year old formalin-fixed paraffin-embedded breast cancer samples: a comparison of column purification and magnetic bead-based technologies

Abstract Background The development of protocols for RNA extraction from paraffin-embedded samples facilitates gene expression studies on archival samples with known clinical outcome. Older samples are particularly valuable because they are associated with longer clinical follow up. RNA extracted from formalin-fixed paraffin-embedded (FFPE) tissue is problematic due to chemical modifications and continued degradation over time. We compared quantity and quality of RNA extracted by four different protocols from 14 ten year old and 14 recently archived (three to ten months old) FFPE breast cancer tissues. Using three spin column purification-based protocols and one magnetic bead-based protocol, total RNA was extracted in triplicate, generating 336 RNA extraction experiments. RNA fragment size was assayed by reverse transcription-polymerase chain reaction (RT-PCR) for the housekeeping gene glucose-6-phosphate dehydrogenase (G6PD), testing primer sets designed to target RNA fragment sizes of 67 bp, 151 bp, and 242 bp. Results Biologically useful RNA (minimum RNA integrity number, RIN, 1.4) was extracted in at least one of three attempts of each protocol in 86–100% of older and 100% of recently archived ("months old") samples. Short RNA fragments up to 151 bp were assayable by RT-PCR for G6PD in all ten year old and months old tissues tested, but none of the ten year old and only 43% of months old samples showed amplification if the targeted fragment was 242 bp. Conclusion All protocols extracted RNA from ten year old FFPE samples with a minimum RIN of 1.4. Gene expression of G6PD could be measured in all samples, old and recent, using RT-PCR primers designed for RNA fragments up to 151 bp. RNA quality from ten year old FFPE samples was similar to that extracted from months old samples, but quantity and success rate were generally higher for the months old group. We preferred the magnetic bead-based protocol because of its speed and higher quantity of extracted RNA, although it produced similar quality RNA to other protocols. If a chosen protocol fails to extract biologically useful RNA from a given sample in a first attempt, another attempt and then another protocol should be tried before excluding the case from molecular analysis.

Storage of bovine reproductive tissues and RNA extracts on ice for 24h or repeated freeze–thaw cycles do not affect RNA integrity

The aims of this study were to test (i) the effect of time of tissue and RNA extracts storage on ice and (ii) the effect of repeated freeze–thaw cycles on RNA integrity and gene expression of bovine reproductive tissues. Fragments of endometrium (ENDO), corpus luteum (CL) and ampulla (AMP) were subdivided and incubated for 0, 1, 3, 6, 12 or 24 h on ice. RNA extracts were incubated on ice for 0, 3, 12 or 24 h, or exposed to 1, 2, 4 or 6 freeze–thaw cycles. RNA integrity number (RIN) was estimated. Expression of progesterone receptor (PGR) and cyclophilin genes from RNA extracts stored on ice for 0 or 24 h, and 1 or 6 freeze–thaw cycles was measured by qPCR. Tissue and RNA extract incubation on ice, and repeated freeze–thaw cycles did not affect RIN values of RNA from ENDO, CL or AMP. Storage on ice or exposure to freeze–thaw cycles did not affect Cq values for PGR or cyclophilin genes. In conclusion, neither generalized RNA degradation nor specific RNA degradation was affected by storage of tissue or RNA extracts on ice for up to 24 h, or by up to 6 freeze–thaw cycles of RNA extracts obtained from bovine ENDO, CL and AMP.