Abstract Pathogenic DNA alterations in GJB2 are present in nearly half of non-syndromic hearing loss cases with autosomal recessive inheritance. The most frequent variant in GJB2 causing non-syndromic hearing loss is the frameshifting c.35del. GJB2 encodes Cx26, a protein of the connexin family that assembles hemichannels and gap junctions. The expression of paralogous proteins is believed to compensate for the loss of function of specific connexins. As Cx26 has been involved in cell differentiation in distinct tissues, we employed stem cells derived from human exfoliated deciduous teeth (SHEDs), homozygous for the c.35del variant, to assess GJB2 roles in stem cell differentiation and the relationship between its loss of function and the expression of paralogous genes. Primary SHED cultures from patients and control individuals were compared. SHEDs from patients had significantly less GJB2 mRNA and increased amount of GJA1 (Cx43), but not GJB6 (Cx30) or GJB3 (Cx31) mRNA. In addition, they presented higher induced differentiation to adipocytes and osteocytes but lower chondrocyte differentiation. Our results suggest that GJA1 increased expression may be involved in functional compensation for GJB2 loss of function in human stem cells, and it may explain changes in differentiation properties observed in SHEDs with and without the c.35del variant. GJB nonsyndromic non syndromic inheritance c35del cdel c 35del del Cx junctions connexins Cx2 tissues SHEDs, , (SHEDs) genes compared GJA Cx43, Cx43 (Cx43) Cx30 (Cx30 Cx31 (Cx31 addition (SHEDs Cx4 (Cx43 Cx3 (Cx3 (Cx4 (Cx

A resposta ao estresse oxidativo não é bem conhecida em plantas como em bactérias, leveduras e humanos. Por exemplo, assume-se que óxido nítrico tem várias funções em plantas apesar do gene que codificaria para óxido nítrico sintetase nunca ter sido isolado. Este trabalho descreve os resultados de uma busca no banco de dados de seqüências expressas de cana de açúcar (SUCEST) de genes envolvidos na resposta ao estresse oxidativo. Eu não encontrei genes similares a óxido nítrico no banco de dados do SUCEST, mas uma via alternativa para a produção deste radical livre pode ser proposta. Eu também encontrei vários genes envolvidos na defesa antioxidante, como quelantes de metais, antioxidantes de baixo peso molecular, enzimas antioxidantes e sistemas de reparo. Ascorbato (vitamina C) é um importante antioxidante em plantas porque é encontrado em altas concentrações em células vegetais e porque é substrato de ascorbato peroxidase, uma enzima que eu encontrei em diferentes isoformas no banco de dados do SUCEST. Eu também encontrei várias enzimas envolvidas na biossíntese de antioxidantes de baixo peso molecular que podem ser alvos para manipulação genética. A obtenção de plantas modificadas geneticamente que sintetizariam vitaminas C e E em altos níveis poderiam melhorar o valor nutricional e a tolerância a estresses de cana de açúcar. Os diversos componentes do sistema de defesa antioxidante interagem entre si e as suas sínteses devem ser muito bem reguladas. Fatores de transcrição envolvidos na regulação da resposta ao estresse oxidativo de bactérias, leveduras e de humanos diferem consideravelmente entre si e quando foram utilizados para buscas no banco de dados do SUCEST, somente genes com similaridades fracas foram encontrados, sugerindo que estas proteínas não são muito conservadas. O envolvimento de espécies reativas de oxigênio e nitrogênio na defesa de plantas contra patógenos também é discutido neste trabalho.

Oxidative stress response in plants is still poorly understood in comparison with the correspondent phenomenon in bacteria, yeast and mammals. For instance, nitric oxide is assumed to play various roles in plants although no nitric oxide synthase gene has yet been isolated. This research reports the results of a search of the sugarcane expressed sequence tag (SUCEST) database for homologous sequences involved in the oxidative stress response. I have not found any gene similar to nitric oxide synthase in the SUCEST database although an alternative pathway for nitric oxide synthesis was proposed. I have also found several genes involved in antioxidant defense, e.g. metal chelators, low molecular weight compounds, antioxidant enzymes and repair systems. Ascorbate (vitamin C) is a key antioxidant in plants because it reaches high concentrations in cells and is a substrate for ascorbate peroxidase, an enzyme that I found in different isoforms in the SUCEST database. I also found many enzymes involved in the biosynthesis of low molecular weight antioxidants, which may be potential targets for genetic manipulation. The engineering of plants for increased vitamin C and E production may lead to improvements in the nutritional value and stress tolerance of sugarcane. The components of the antioxidant defense system interact and their synthesis is probably closely regulated. Transcription factors involved in regulation of the oxidative stress response in bacteria, yeast and mammals differ considerably among themselves and when I used them to search the SUCEST database only genes with weak similarities were found, suggesting that these transcription regulators are not very conserved. The involvement of reactive oxygen species and antioxidants in plant defense against pathogens is also discussed.