Multi-fluorescent silica colloids for encoding large combinatorial libraries

Large chemical libraries can be synthesized on solid-support beads by the combinatorial split-and-mix method. A major challenge associated with this type of library synthesis is distinguishing between the beads and their attached compounds. A new method of encoding these solid-support beads, 'colloidal bar-coding', involves attaching fluorescent silica colloids ('reporters') to the beads as they pass through the compound synthesis, thereby creating a fluorescent bar code on each bead. In order to obtain sufficient reporter varieties to bar code extremely large libraries, many of the reporters must contain multiple fluorescent dyes. We describe here the synthesis and spectroscopic analysis of various mono- and multi-fluorescent silica particles for this purpose. It was found that by increasing the amount of a single dye introduced into the particle reaction mixture, mono- fluorescent silica particles of increasing intensities could be prepared. This increase was highly reproducible and was observed for six different fluorescent dyes. Multi-fluorescent silica particles containing up to six fluorescent dyes were also prepared. The resultant emission intensity of each dye in the multi-fluorescent particles was found to be dependent upon a number of factors; the hydrolysis rate of each silane-dye conjugate, the magnitude of the inherent emission intensity of each dye within the silica matrix, and energy transfer effects between dyes. We show that by varying the relative concentration of each silane-dye conjugate in the synthesis of multi-fluorescent particles, it is possible to change and optimize the resultant emission intensity of each dye to enable viewing in a fluorescence detection instrument.

The development and application of a novel safety-catch linker for BOC-based assembly of libraries of cyclic peptides

Cyclic peptides are appealing targets in the drug-discovery process. Unfortunately, there currently exist no robust solid-phase strategies that allow the synthesis of large arrays of discrete cyclic peptides. Existing strategies are complicated, when synthesizing large libraries, by the extensive workup that is required to extract the cyclic product from the deprotection/cleavage mixture. To overcome this, we have developed a new safety-catch linker. The safety-catch concept described here involves the use of a protected catechol derivative in which one of the hydroxyls is masked with a benzyl group during peptide synthesis, thus making the linker deactivated to aminolysis. This masked derivative of the linker allows BOC solid-phase peptide assembly of the linear precursor. Prior to cyclization, the linker is activated and the linear peptide deprotected using conditions commonly employed (TFMSA), resulting in deprotected peptide attached to the activated form of the linker. Scavengers and deprotection adducts are removed by simple washing and filtration. Upon neutralization of the N-terminal amine, cyclization with concomitant cleavage from the resin yields the cyclic peptide in DMF solution. Workup is simple solvent removal. To exemplify this strategy, several cyclic peptides were synthesized targeted toward the somatostatin and integrin receptors. From this initial study and to show the strength of this method, we were able to synthesize a cyclic-peptide library containing over 400 members. This linker technology provides a new solid-phase avenue to access large arrays of cyclic peptides.

The effect of mode of exposure to Beauveria bassiana on conidia acquisition and host mortality of Colorado potato beetle, Leptinotarsa decemlineata

The effects of the mode of exposure of second instar Colorado potato beetles to Beauveria bassiana on conidia acquisition and resulting mortality were investigated in laboratory studies. Larvae sprayed directly with a B, bassiana condial suspension, larvae exposed to B, bassiana-treated foliage, and larvae both sprayed and exposed to treated foliage experienced 76, 34, and 77% mortality, respectively. The total number of conidia and the proportion of germinating conidia were measured over time for four sections of the insect body: the ventral surface of the head (consisting mostly of ventral mouth parts), the ventral abdominal surface, the dorsal abdominal surface, and the legs. From observations at 24 and 36 h posttreatment, mean totals of 161.1 conidia per insect were found on sprayed larvae, 256.1 conidia on larvae exposed only to treated foliage, and 408.3 conidia on larvae both sprayed and exposed to treated foliage, On sprayed larvae, the majority of conidia were found on the dorsal abdominal surface, whereas conidia were predominantly found in the ventral abdominal surface and mouth parts on larvae exposed to treated foliage, Between 24 and 36 h postinoculation the percentage of conidia germinating on sprayed larvae increased slightly from 80 to 84%), On the treated foliage, the percentage of germinated conidia on larvae increased from 35% at 24 h to 50% at 36 h posttreatment, Conidia germination on sprayed larvae on treated foliage was 65% at 24 h and 75% at 36 h posttreatment, It is likely that the gradual acquisition of conidia derived from the continuous exposure to B. bassiana inoculum on the foliar surface was responsible for the increase in germination over time on larvae exposed to treated foliage, The density and germination of conidia were observed 0, 4, 8, 12, 16, 20, and 24 h after being sprayed with or dipped in conidia suspensions or exposing insects to contaminated foliage, Conidia germinated twice as fast on sprayed insects as with any other treatment within the first 12 h, This faster germination may be due to the pressure of the sprayer enhancing conidial lodging on cuticular surfaces. (C) 2001 Academic Press.

Evaluation of synergistic interactions between the Colorado potato beetle (Coleoptera : Chrysomelidae) pathogen Beauveria bassiana and the insecticides, imidacloprid, and cyromazine

Laboratory studies investigated the interaction between the fungal entomopathogen Beauveria bassiana (Balsamo) Vuillemin and sublethal doses of the insecticides imidacloprid and cyromazine when applied to larvae of the Colorado potato beetle, Leptinotarsa decemlinenta (Say). When second instars were fed potato leaf discs treated with sublethal doses of imidacloprid and a range of doses of B. bassiana, a synergistic action was demonstrated. Similar results were observed when larvae were sprayed directly with B. bassiana conidia and immediately fed leaf discs treated with imidacloprid. No synergistic interaction was detected when larvae were fed leaf discs treated with sublethal doses of imidacloprid 24 h after application of R. bassiana conidia to larvae. However, a synergistic interaction was detected when larvae were fed leaf discs treated with imidacloprid and sprayed with B, bassiana conidia 24 h later. Although sublethal doses of both imidacloprid and the triazine insect growth regulator (IGR) cyromazine prolonged the duration of the second instar, only imidacloprid interacted with B. bassiana to produce a synergistic response in larval mortality. In leaf consumption studies, the highest dose of B, bassiana tested promoted feeding in inoculated second instars. Feeding was inhibited when larvae were fed foliage treated with sublethal doses of imidacloprid and significantly reduced when fed foliage treated with a sublethal dose of cyromazine. Starvation of larvae for 24 h immediately after B. bassiana treatment produced a similar result to the combined treatment of B. bassiana and imidacloprid and increased the level of mycosis when compared with B. bassiana controls. Imidacloprid treatment affected neither the rate of germination of B. bassiana conidia on the insect cuticle nor the rate at which conidia were removed from the integument after application. The statistical analysis used to detect synergism and the possible role of starvation-induced stress factors underlying the observed synergistic interactions are discussed.

A subset-oriented algorithm for minimizing the number of steps required for synthesis of cyclic-peptide libraries

Libraries of cyclic peptides are being synthesized using combinatorial chemistry for high throughput screening in the drug discovery process. This paper describes the min_syn_steps.cpp program (available at http://www.imb.uq.edu.au/groups/smythe/tran), which after inputting a list of cyclic peptides to be synthesized, removes cyclic redundant sequences and calculates synthetic strategies which minimize the synthetic steps as well as the reagent requirements. The synthetic steps and reagent requirements could be minimized by finding common subsets within the sequences for block synthesis. Since a brute-force approach to search for optimum synthetic strategies is impractically large, a subset-orientated approach is utilized here to limit the size of the search. (C) 2002 Elsevier Science Ltd. All rights reserved.

Desenvolvimento e qualidade da lima ácida Tahiti em Colorado do Oeste, RO

Os aspectos de crescimento, desenvolvimento e qualidade de frutos cítricos na região de Colorado do Oeste, RO, não têm sido explorados. Dessa forma, o objetivo deste trabalho foi avaliar o crescimento e a qualidade fisico-química de frutos de lima ácida Tahiti em diferentes épocas de desenvolvimento, sem irrigação, na região de Colorado do Oeste, RO. O experimento foi realizado no Instituto Federal de Rondônia (IFRO), Campus de Colorado do Oeste, durante os anos de 2006, 2007 e 2008, com o acompanhamento das safras de 2006/07 e 2007/08. Para cada safra foram avaliadas oito épocas de desenvolvimento dos frutos, da fase de chumbinho à colheita. Determinaram-se soma térmica em graus-dia, unidades fototérmicas e precipitação pluviométrica. Houve influência das condições meteorológicas na duração das épocas de desenvolvimento dos frutos e na qualidade dos frutos na safra 2006/07, mas na safra 2007/08 não houve diferença significativa para peso, tamanho médio dos frutos, acidez e rendimento médio de suco. Tamanho e peso dos frutos, pH, sólidos solúveis, acidez, relação SS/AT e teor de vitamina C do suco dos frutos apresentaram valores médios adequados aos padrões comerciais exigidos pelo mercado in natura e pela indústria. O rendimento de suco dos frutos colhidos na safra 2006/07 apresentou qualidade satisfatória para atender às necessidades do mercado in natura ou industrial, mas não para a safra 2007/08, devido ao baixo rendimento de suco dos frutos, provavelmente em razão da menor precipitação pluviométrica ocorrida nessa safra. O índice tecnológico encontrado nas safras avaliadas está abaixo do exigido pela indústria.

Developing Libraries Using Software Transactional Memory

Software transactional memory is a promising programming model that adapts many concepts borrowed from the databases world to control concurrent accesses to main memory (RAM) locations. This paper discusses how to support apparently irreversible operations, such as memory allocation and deallocation, within software libraries that will be used in (software memory) transactional contexts, and propose a generic and elegant approach based on a handler system, which provide the means to create and execute compensation actions at key moments during the life-time of a transaction.

Estoque de nutrientes sob diferentes sistemas de uso do solo de Colorado do Oeste-RO

A substituição da floresta por outro tipo de uso do solo pode levar a perdas significativas na matéria orgânica do solo, alterando sua dinâmica e como consequência alterando as entradas e saídas de nutrientes do sistema. Neste trabalho o objetivo foi avaliar o estoque de nutrientes em diferentes sistemas de uso de solo em Colorado do Oeste-RO. Os sistemas avaliados foram o agroflorestal (teca e cacau), florestal (teca com cinco anos), teca com oito anos, agrossilvopastoril (teca, cacau e pasto) e pastagem tendo a mata nativa como referência. Em cada sistema foram abertas três minitrincheiras, nas quais foram coletadas amostras deformadas e indeformadas nas profundidades de 0 a 5, 5 a 10, 10 a 20 e 20 a 30 cm. Nessas amostras foram determinados os nutrientes: N, P, K, Ca e Mg, a matéria orgânica, a densidade do solo e os teores de argila. Observaram-se diferenças nos estoques de nutrientes no solo entre os sistemas de uso e em profundidade. A concentração de nutrientes foi influenciada pelo sistema de uso do solo. O nutriente mais estocado em todos os sistemas de uso é o Ca, os valores variaram entre 5188,48 e 2912,86 kg ha-1 e o menos estocado o K, com estoques entre 46,22 e 17,33 kg ha-1. O sistema teca com cinco anos foi o sistema que mais se aproximou da mata, quanto ao armazenamento de nutrientes.

Flora of the Rocky Mountains and adjacent plains, Colorado, Utah, Wyoming, Idaho, Montana, Saskatchewan, Alberta, and neighboring parts of Nebraska, South Dakota, and British Columbia /by P. A. Rydberg...

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