Summary Created in 2007, COMUSA is a multiprofessional committee comprising speech therapy, otology, otorhinolaryngology and pediatrics with the aim of debating and countersigning auditory health actions for neonatal, lactating, preschool and school children, adolescents, adults and elderly persons. COMUSA includes representatives of the Brazilian Audiology Academy (Academia Brasileira de Audiologia or ABA), the Brazilian Otorhinolaryngology and Cervicofacial Surgery Association (Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico Facial or ABORL), the Brazilian Phonoaudiology Society (Sociedade Brasileira de Fonoaudiologia or SBFa), the Brazilian Otology Society (Sociedade Brasileira de Otologia or SBO), and the Brazilian Pediatrics Society (Sociedade Brasileira de Pediatria or SBP).

We use the Box-Jenkins approach to fit an autoregressive integrated moving average (ARIMA) model to dengue incidence in Rio de Janeiro, Brazil, from 1997 to 2004. We find that the number of dengue cases in a month can be estimated by the number of dengue cases occurring one, two, and twelve months prior. We use our fitted model to predict dengue incidence for the year 2005 when two alternative approaches are used: 12-steps ahead versus 1-step ahead. Our calculations show that the 1-step ahead approach for predicting dengue incidence provides significantly more accurate predictions (P value=0.002, Wilcoxon signed-ranks test) than the 12-steps ahead approach. We also explore the predictive power of alternative ARIMA models incorporating climate variables as external regressors. Our findings indicate that ARIMA models are useful tools for monitoring dengue incidence in Rio de Janeiro. Furthermore, these models can be applied to surveillance data for predicting trends in dengue incidence.

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Plants of the genus Polygala have been shown to possess protective effects against neuronal death and cognitive impairments in neurodegenerative disorders related to excitotoxicity. Moreover, previous reports from our group have shown the neuroprotective effects of the plant Polygala paniculata against methylmercury (MeHg)-induced neurotoxicity. In this work, we have examined the potential protective effects of three compounds (7-prenyloxy-6-methoxycoumarin, quercetin, and 1,5-dihidroxi-2,3-dimethoxy xanthone) from Polygala species against MeHg- and mercuric chloride (HgCl2)-induced disruption of mitochondrial function under in vitro conditions using mitochondrial-enriched fractions from mouse brain. MeHg and HgCl2 (10-100 microM) significantly decreased mitochondrial viability; this phenomenon was positively correlated to mercurial-induced glutathione oxidation. Among the isolated compounds, only quercetin (100-300 microM) prevented mercurial-induced disruption of mitochondrial viability. Moreover, quercetin, which did not display any chelating effect on MeHg or HgCl2, prevented mercurial-induced glutathione oxidation. The present results suggest that the protective effects of quercetin against mercurial-induced mitochondrial dysfunction is related to the removal of oxidant species generated in the presence of either MeHg or HgCl2. Reinforcing this hypothesis, MeHg and HgCl2 increased the production of hydrogen peroxide in the brain mitochondria, as well as the levels of malondialdehyde. These oxidative phenomena were prevented by co-incubation with quercetin or catalase. These results are the first to show the involvement of hydrogen peroxide as a crucial molecule related to the toxic effects of both organic and inorganic mercurials in brain mitochondria. In addition, the study is the first to show the protective effect of quercetin against mercurial-induced toxicity, pointing to its capability to counteract mercurial-dependent hydrogen peroxide generation as a potential molecular mechanism of protection. Taken together, these data render quercetin a promising molecule for pharmacological studies with respects to mercurials' poisoning.

Citrus canker, caused by Xanthomonas axonopodis Starr and Garces pv. citri (Hasse) Vauterin et al., is one of the main problems affecting citrus production. In order to obtain resistance to phytopathogenic bacteria, insect genes, coding for antimicrobial proteins, have been used in plant genetic transformation. In this study, transgenic Citrus sinensis (L.) Osb. `Hamlin' plants expressing the antimicrobial insect-derived attacin A gene (attA) were obtained by Agrobacterium tumefaciens (Smith and Towns.) Conn-mediated transformation. Initially, the cDNA clone was used to construct a binary plasmid vector (pCattA 2300). The construction included the native signal peptide (SP) responsible for directing the insect protein to the extracellular space where bacteria is supposed to accumulate in vivo. In order to investigate the native SP effectiveness in a plant model system, onion (Allium cepa L.) epidermal cells were transformed, via biobalistics, using plasmids containing the attA gene with or without SP, fused with the green fluorescent protein gene (pattA 1303 and pSPattA 1303). Fluorescence accumulation surrounding the cells was observed only in tissues transformed with the plasmid containing the gene with SP, indicating the protein secretion to the apoplast. Citrus transformation was confirmed by PCR and Southern blot hybridization analysis in 12 regenerated plants. Transcription of attA gene was detected by Northern blot analysis in all transgenic plants. Eight selected transgenic lines were propagated and inoculated with a 106 cfu/mL suspension of the pathogen X. axonopodis pv. citri. Compared to control (non-transformed plant), seven transgenic lines showed a significant reduction in susceptibility to citrus canker. The results obtained here indicate the potential use of antibacterial proteins to protect citrus from bacterial diseases.

A new method for obtaining transgenic sweet orange plants was developed in which positive selection (Positech) based on the Escherichia coli phosphomannose-isomerase (PMI) gene as the selectable marker gene and mannose as the selective agent was used. Epicotyl segments from in vitro-germinated plants of Valencia, Hamlin, Natal and Pera sweet oranges were inoculated with Agrobacterium tumefaciens EHA101-pNOV2116 and subsequently selected on medium supplemented with different concentrations of mannose or with a combination of mannose and sucrose as a carbon source. Genetic transformation was confirmed by PCR and Southern blot. The transgene expression was evaluated using a chlorophenol red assay and isoenzymes. The transformation efficiency rate ranged from 3% to 23.8%, depending on cultivar. This system provides an efficient manner for selecting transgenic sweet orange plants without using antibiotics or herbicides.

This work reports the evaluation of citrus canker resistance in transgenic lines of four sweet orange cultivars Hamlin, Natal, Pera and Valencia expressing the rice Xa21 R-gene. The Agrobacterium-mediated genetic transformation was performed and transgene integration was confirmed by Southern blot analyses. These analyses showed that the plants had undergone 1-3 independent transgene integration events. The genetic transformation efficiency was 18AE6% for Hamlin, 8AE3% for Natal, 3% for Pera and 11AE3% for Valencia sweet orange. Transgene transcription was confirmed by northern blot analyses in most transgenic lines analysed. Transgenic lines of each of the cultivars were selected and propagated for resistance evaluation to Xanthomonas axonopodis pv. citri. Transgenic lines with a higher resistance to citrus canker were identified for cvs Hamlin, Natal and Pera. The lowest diseased leaf area values were detected for transgenic lines of Natal and Pera, cultivars which are considered moderately resistant to citrus canker in the field, indicating an influence of cultivar genetic background on the effectiveness of the Xa21 gene in controlling the pathogen. Transgenic lines in which transgene transcription was not detected did not show any improvement in resistance to the pathogen. These results show the integration and expression of the rice Xa21 gene in citrus and the possibility of using R-genes in controlling diseases in a heterologous genetic background.

Somatic embryogenesis from nucellus-derived callus cultures of five cultivars, including three (Caipira, Seleta Vermelha, and Valencia) of sweet oranges (C. sinensis L. Osbeck), Rangpur lime (C. limonia L. Osbeck), and Cleopatra mandarin (C. reticulata Blanco) (lines I and II), were studied. Callus lines maintained on MT medium supplemented with 50 g l 21 sucrose were transferred to MT medium supplemented with different carbohydrate sources: galactose, glucose, lactose, maltose, or sucrose at 18, 37, 75, 110, or 150 mM, or glycerol at 6, 12, 24, 36, or 50 mM. Globular embryos were observed after approximately 4 wk, in several treatments. Cultures of Valencia and Caipira sweet oranges and Cleopatra mandarin (line I) showed high numbers of embryos on medium containing galactose, lactose, and maltose. Histological studies showed somatic embryos in all developmental stages with a normal histodifferentiation pattern. The other two cultivars (Rangpur lime and Cleopatra mandarin, line II) formed very few embryos, which did not develop further following the globular stage. Some of the abnormalities observed were lack or dedifferentiation of protoderm and absence of apical meristems and procambial strands. Embryos that followed the normal sequence of development were easily converted into plants. Non-embryogenic cultures continued as proliferating callus cultures, eventually forming a few embryos which did not convert into plants. Statistical analyses of the callus response to carbohydrate treatments was done using an overdispersion Poisson model.