Stoichiometric and irreversible cysteine-selective protein modification using carbonylacrylic reagents

Maleimides remain the reagents of choice for the preparation of therapeutic and imaging protein conjugates despite the known instability of the resulting products that undergo thiol-exchange reactions $\textit{in vivo}$. Here we present the rational design of carbonylacrylic reagents for chemoselective cysteine bioconjugation. These reagents undergo rapid thiol Michael-addition under biocompatible conditions in stoichiometric amounts. When using carbonylacrylic reagents equipped with PEG or fluorophore moieties, this method enables access to protein and antibody conjugates precisely modified at pre-determined sites. Importantly, the conjugates formed are resistant to degradation in plasma and are biologically functional, as demonstrated by the selective imaging and detection of apoptotic and HER2+ cells, respectively. The straightforward preparation, stoichiometric use and exquisite cysteine selectivity of the carbonylacrylic reagents combined with the stability of the products and the availability of biologically relevant cysteine-tagged proteins make this method suitable for the routine preparation of chemically defined conjugates for $\textit{in vivo}$ applications.FAPESP (Grant IDs: 2012/22274-2; BEPE 2015/07509-1, 2013/25504-1), Xunta de Galicia, FCT Portugal (FCT Investigator, SFRH/BPD/103172/2014 Postdoctoral fellowship, SFRH/BD/111556/2015 PhD Studentship), European Union (Marie-Sklodowska Curie ITN Protein Conjugates), Engineering and Physical Sciences Research Council, MECD (‘Salvador Madariaga’ mobility grant PRX15/00638), MINECO (CTQ2015-70524-R, RYC-2013-14706 ), Royal Society, European Research Council Starting Grant (TagIt

Qualidade de alface crespa cultivada em sistema orgânico, convencional e hidropônico.

O objetivo deste trabalho foi avaliar a qualidade da alface do grupo crespa, cv. Vera, em sistemas de cultivo orgânico, convencional e hidropônico em Rio Branco-AC. O experimento foi conduzido em julho de 2009. As amostras dos sistemas, convencional e hidropônico (3 marcas comerciais), foram escolhidas aleatoriamente nos supermercados do município de Rio Branco, no mesmo dia de coleta da alface produzida em sistema orgânico. A alface orgânica, produzida na área experimental do Setor de Agricultura Ecológica da Universidade Federal do Acre (UFAC), em Rio Branco, foi cultivada em estufa, sob plantio direto utilizando folhas de bambu como cobertura do solo, e adubada com composto orgânico (17 t ha-1 em base seca). O delineamento utilizado foi o inteiramente casualizado, com cinco tratamentos e quatro repetições compostas por três plantas. As amostras foram lavadas, cortadas e processadas com folha e caule, para obter o suco. Logo após foram determinados os teores de sólidos solúveis e a concentração de nitrato e ácido ascórbico. As três marcas de alface hidropônica apresentaram maior teor de nitrato e menor concentração de sólidos solúveis e ácidos ascórbico, enquanto a alface orgânica apresentou qualidade superior, com baixa concentração de nitrato e maior teor de ácido ascórbico

Combining ability of summer-squash lines with different degrees of parthenocarpy and PRSV-W resistance

The aim was to assess heterosis in a set of 16 summer-squash hybrids, and evaluate the combining capacity of the respective parental lines, which differed as to the degree of parthenocarpy and resistance to PRSV-W (Papaya Ringspot Virus-Watermelon strain). The hybrids were obtained using a partial diallel cross design (4 × 4). The lines of parental group I were 1 = ABX-037G-77-03-05-01-01-bulk, 2 = ABX-037G-77-03-05-03-10-bulk, 3 = ABX-037G-77-03-05-01-04-bulk and 4 = ABX-037G-77-03-05-05-01-bulk, and of group II, 1′ = ABX-037G-77-03-05-04-08-bulk, 2′ = ABX-037G-77-03-05-02-11-bulk, 3′ = Clarice and 4′ = Caserta. The 16 hybrids and eight parental lines were evaluated for PRSV-W resistance, parthenocarpic expression and yield in randomized complete-block designs, with three replications. Parthenocarpy and the resistance to PRSV-W were rated by means of a scale from 1 to 5, where 1 = non-parthenocarpic or high resistance to PRSV-W, and 5 = parthenocarpic or high susceptibility to PRSV-W. Both additive and non-additive gene effects were important in the expression of parthenocarpy and resistance to PRSV-W. Whereas estimates of heterosis in parthenocarpy usually tended towards a higher degree, resistance to PRSV-W was towards higher susceptibility. At least one F1 hybrid was identified with a satisfactory degree of parthenocarpy, resistance to PRSV-W and high fruit-yield

Identification of new Bremia lactucae races in lettuce in São Paulo state

Lettuce is the most consumed leafy vegetable in Brazil. However, its production has faced increasing difficulties mainly due to the infestation of producing areas with Bremia lactucae, the causing agent of downy mildew, the worst disease in lettuce. The aim of this study was to identify B. lactucae races occurring in lettuce producing areas of São Paulo state, during 2008 and 2009. Lettuce leaf samples containing B. lactucae sporangia were collected in the major lettuce producing areas of São Paulo. Each sample was considered an isolate. After the multiplication of sporangia in the susceptible cultivar Solaris, with subsequent inoculation in the differentiating cultivars, the evaluations were performed on the same day of the first sporulation in the susceptible cultivar 'Cobham Green (DM 0)'. In 2008 one new B. lactucae code was identified. SPBl:05 was the denomination proposed to it. In 2009 one another B. lactucae code was identified, that is, 63/63/02/00, whose denomination was proposed to be SPBl:06.A alface é o grupo de hortaliças folhosas de maior consumo no Brasil. No entanto a dificuldade em produzi-la vem aumentando, principalmente pela infestação das áreas de produção por Bremia lactucae, agente causador do míldio, principal doença da alface. O objetivo deste trabalho foi identificar as raças de B. lactucae que ocorreram nos municípios produtores de alface do Estado de São Paulo nos anos de 2008 e 2009. Para isso, coletaram-se folhas com esporângios de B. lactucae em municípios produtores de alface, sendo que cada amostra foi considerada um isolado. Após a multiplicação dos esporângios na cultivar suscetível Solaris, com posterior inoculação nas diferenciadoras, realizaram-se as avaliações, no mesmo dia do aparecimento da primeira esporulação na cultivar suscetível 'Cobham Green (DM 0)'. No ano 2008 identificou-se um novo código de B. lactucae em alface, à qual se propôs a denominação de SPBl:05. No ano 2009, outro código de B. lactucae foi identificado(63/63/02/00), ao qual se propôs a denominação de SPBl:06