Identification of Pathogenicity-Related Genes in the Vascular Wilt Fungus Verticillium dahliae by Agrobacterium tumefaciens-Mediated T-DNA Insertional Mutagenesis

Verticillium dahliae is the causal agent of vascular wilt in many economically important crops worldwide. Identification of genes that control pathogenicity or virulence may suggest targets for alternative control methods for this fungus. In this study, Agrobacteriumtumefaciens-mediated transformation (ATMT) was applied for insertional mutagenesis of V. dahliae conidia. Southern blot analysis indicated that T-DNAs were inserted randomly into the V. dahliae genome and that 69% of the transformants were the result of single copy T-DNA insertion. DNA sequences flanking T-DNA insertion were isolated through inverse PCR (iPCR), and these sequences were aligned to the genome sequence to identify the genomic position of insertion. V. dahliae mutants of particular interest selected based on culture phenotypes included those that had lost the ability to form microsclerotia and subsequently used for virulence assay. Based on the virulence assay of 181 transformants, we identified several mutant strains of V. dahliae that did not cause symptoms on lettuce plants. Among these mutants, T-DNA was inserted in genes encoding an endoglucanase 1 (VdEg-1), a hydroxyl-methyl glutaryl-CoA synthase (VdHMGS), a major facilitator superfamily 1 (VdMFS1), and a glycosylphosphatidylinositol (GPI) mannosyltransferase 3 (VdGPIM3). These results suggest that ATMT can effectively be used to identify genes associated with pathogenicity and other functions in V. dahliae

Recursive splicing in long vertebrate genes

It is generally believed that splicing removes introns as single units from precursor messenger RNA transcripts. However, some long Drosophila melanogaster introns contain a cryptic site, known as a recursive splice site (RS-site), that enables a multi-step process of intron removal termed recursive splicing. The extent to which recursive splicing occurs in other species and its mechanistic basis have not been examined. Here we identify highly conserved RS-sites in genes expressed in the mammalian brain that encode proteins functioning in neuronal development. Moreover, the RS-sites are found in some of the longest introns across vertebrates. We find that vertebrate recursive splicing requires initial definition of an RS-exon that follows the RS-site. The RS-exon is then excluded from the dominant mRNA isoform owing to competition with a reconstituted 5 splice site formed at the RS-site after the first splicing step. Conversely, the RS-exon is included when preceded by cryptic promoters or exons that fail to reconstitute an efficient 5 splice site. Most RS-exons contain a premature stop codon such that their inclusion can decrease mRNA stability. Thus, by establishing a binary splicing switch, RS-sites demarcate different mRNA isoforms emerging from long genes by coupling cryptic elements with inclusion of RS-exons

Processamento auditivo: comparação entre potenciais evocados auditivos de média latência e testes de padrões temporais Auditory processing: comparision between auditory middle latency response and temporal pattern tests

OBJETIVO: verificar a concordância entre os resultados da avaliação do Potencial Evocado Auditivo de Média Latência e testes de padrões temporais. MÉTODOS: foram avaliados 155 sujeitos de ambos os sexos, idade entre sete e 16 anos, com audição periférica normal. Os sujeitos foram submetidos aos testes de Padrão de Frequência e Duração e Potenciais Evocados auditivos de Média Latência. RESULTADOS: os sujeitos foram distribuídos em dois grupos: normal ou alterado para o processamento auditivo. O índice de alteração foi em torno de 30%, exceto para Potencial Evocado Auditivo de Média Latência que foi pouco menor (17,4%). Os padrões de frequência e duração foram concordantes até 12 anos. A partir dos 13 anos, observou-se maior ocorrência de alteração no padrão de frequência que no padrão de duração. Os padrões de frequência e duração (orelhas direita e esquerda) e Potencial Evocado Auditivo de Média Latência não foram concordantes. Para 7 e 8 anos a combinação padrão de frequência e duração normal / Média Latência alterado tem maior ocorrência que a combinação padrão de frequência e duração alterada / Média Latência normal. Nas demais idades, ocorreu o contrário. Não houve diferença estatística entre as faixas etárias quanto à distribuição de normal e alterado no padrão de frequência (orelhas direita e esquerda), nem para o Potencial Evocado Auditivo de Média Latência, com exceção do padrão de duração para o grupo de 9 e 10 anos. CONCLUSÃO: não houve concordância entre os resultados do Potencial Evocado Auditivo de Média Latência e os testes de padrões temporais aplicados.<br>PURPOSE: to check the concordance between the Middle Latency Response and temporal processing tests. METHODS: 155 normal hearing subjects of both genders (age group range between 7 to 16 years) were evaluated with the Pitch and Duration Pattern Tests (behavioral) and Middle Latency Response (electrophysiologic) and divided into two groups: normal and abnormal, according to their test results. RESULTS: among all subjects, 30% showed abnormality in the tests, except for the Middle Latency Response that was under 17.4%. The pitch and duration patterns (right and left ears) agreed until 12 years of age. From 13 years, there was a greater number of alteration in the pitch patterns than in the duration patterns. The pitch and duration patterns (right and left ears) and MLR did not show concordance. For the 7 and 8-year-old group, the combination pitch and duration patterns normal/abnormal Middle Latency Response had greater occurrence than the combination pitch and duration patterns abnormal / normal Middle Latency Response. For the other groups the opposite occurred. There was not statistical difference among the age groups regarding normal and abnormal results for the pitch patterns (right and left ears) and Middle Latency Response, the duration patterns showed more abnormal results for the 9, 10 years old group. CONCLUSION: it was not possible to verify concordance between the Middle Latency Response and behavioral evaluation of Frequency and Duration Patterns Test

Pharmacometrics Markup Language (PharmML): Opening New Perspectives for Model Exchange in Drug Development

The lack of a common exchange format for mathematical models in pharmacometrics has been a long-standing problem. Such a format has the potential to increase productivity and analysis quality, simplify the handling of complex workflows, ensure reproducibility of research, and facilitate the reuse of existing model resources. Pharmacometrics Markup Language (PharmML), currently under development by the Drug Disease Model Resources (DDMoRe) consortium, is intended to become an exchange standard in pharmacometrics by providing means to encode models, trial designs, and modeling steps

Pharmacometrics Markup Language (PharmML): Opening New Perspectives for Model Exchange in Drug Development

The lack of a common exchange format for mathematical models in pharmacometrics has been a long-standing problem. Such a format has the potential to increase productivity and analysis quality, simplify the handling of complex workflows, ensure reproducibility of research, and facilitate the reuse of existing model resources. Pharmacometrics Markup Language (PharmML), currently under development by the Drug Disease Model Resources (DDMoRe) consortium, is intended to become an exchange standard in pharmacometrics by providing means to encode models, trial designs, and modeling steps