5 resultados para Phase 1 xenobiotic-metabolizing gene Cyp2a5
em Brock University, Canada
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Phase 1 Map of the Master Plan showing the Tower and Thistle Complex, along with some additional buildings and the main parking lot. Note the location of the Central Utilities Building to the east differs from it's actual location along the escarpment.
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The objectives of the present study were to explore three components of organizational commitment (affective [AC], normative [NC] and continuance [CC] commitment; Allen & Meyer, 1991), perceived relatedness (Oeci & Ryan, 1985; 2002), and behavioural intention (Ajzen, 2002) within the context of volunteer track and field officiating. The objectives were examined in a 2-phase study. Ouring phase 1, experts (N = 10) with domain familiarity assessed the item content relevance and representation of modified organizational commitment (OC; Meyer, Allen & Smith, 1993) and perceived relatedness (La Guardia, Oeci, Ryan & Couchman, 2000) items. Fourteen of 26 (p < .05) items were relevant (Aiken's coefficient V) and NC (M = 3.88, SO = .64), CC (M = 3.63, SD = .52), and relatedness (M = 4.00, SD = .93) items had mean item content-representation ratings of either "good" or "very good" while AC (M = 2.50, SD = 0.58) was rated "fair". Participants in phase 2 (N = 80) responded to items measuring demographic variables, perceptions of OC to Athletics Canada, perceived relatedness to other track and field officials, and a measure of intention (yiu, Au & Tang, 2001) to continue officiating. Internal consistency reliability estimates (Cronbach's (1951) coefficient alpha) were as follows: (a) AC = .78, (b) CC = .85, (c) NC = .80 (d) perceived relatedness = .70 and, (e) intention = .92 in the present sample. Results suggest that the track and field officials felt only minimally committed to Athletics Canada (AC M = 3.90, SD = 1.23; NC M = 2.47, SD = 1.25; CC M = 3.32; SD = 1.34) and that their relationships with other track and field officials were strongly endorsed (M = 5.86, SD = 0.74). Bivariate correlations (Pearson r) indicated that perceived relatedness to other track and field officials demonstrated the strongest relationship with intention to continue officiating (r = .346, p < .05), while dimensions of OC were not significantly related to intention (all p's > .05). Together perceived relatedness (j3 = .339, p = .004), affective commitment (j3 = -.1 53, p = .308), normative commitment (j3 = -.024, p = .864) and continuance commitment (j3 = .186, P = .287) contribute to the prediction of intention to continued officiating (K = .139). These relationships remained unaffected by the inclusion of demographic (j3age = -.02; P years with Athletics Canada = -.13; bothp's > .05) or alternative commitment (j3sport = -.19; P role = .15; Pathletes = .20; all p' s > .05) considerations. Three open-ended questions elicited qualitative responses regarding participants' reasons for officiating. Responses reflecting initial reasons for officiating formed these higher order themes: convenience, helping reasons, extension of role, and intrinsic reasons. Responses reflecting reasons for continuing to officiate formed these higher order themes: track and field, to help, and personal benefits. Responses reflecting changes that would influence continued involvement were: political, organizational/structural, and personal. These results corroborate the findings of previous investigations which state that the reasons underpinning volunteer motivations change over time (Cuskelly et al., 2002). Overall, the results of this study suggest that track and field officials feel minimal commitment to the organization of Athletics Canada but a stronger bond with their fellow officials. Moreover, the degree to which track and field officials feel meaningfully connected to one another appears to exert a positive influence on their intentions to continue officiating. As such, it is suggested that in order to promote continued involvement, Athletics Canada increases its focus on fostering environments promoting positive interactions among officials.
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Agaricus bisporus is the most commonly cultivated mushroom in North America and has a great economic value. Green mould is a serious disease of A. bisporus and causes major reductions in mushroom crop production. The causative agent of green mould disease in North America was identified as Trichoderma aggressivum f. aggressivum. Variations in the disease resistance have been shown in the different commercial mushroom strains. The purpose of this study is to continue investigations of the interactions between T. aggressivum and A. bisporus during the development of green mould disease. The main focus of the research was to study the roles of cell wall degrading enzymes in green mould disease resistance and pathogenesis. First, we tried to isolate and sequence the N-acetylglucosaminidase from A. bisporus to understand the defensive mechanism of mushroom against the disease. However, the lack of genomic and proteomic information of A. bisporus limited our efforts. Next, T. aggressivum cell wall degrading enzymes that are thought to attack Agaricus and mediate the disease development were examined. The three cell wall degrading enzymes genes, encoding endochitinase (ech42), glucanase (fJ-1,3 glucanase) and protease (prb 1), were isolated and sequenced from T. aggressivum f. aggressivum. The sequence data showed significant homology with the corresponding genes from other fungi including Trichoderma species. The transcription levels of the three T. aggressivum cell wall degrading enzymes were studied during the in vitro co-cultivation with A. bisporus using R T -qPCR. The transcription levels of the three genes were significantly upregulated compared to the solitary culture levels but were upregulated to a lesser extent in co-cultivation with a resistant strain of A. bisporus than with a sensitive strain. An Agrobacterium tumefaciens transformation system was developed for T. aggressivum and was used to transform three silencing plasmids to construct three new T. aggressivum phenotypes, each with a silenced cell wall degrading enzyme. The silencing efficiency was determined by RT-qPCR during the individual in vitro cocultivation of each of the new phenotypes with A. bisporus. The results showed that the expression of the three enzymes was significantly decreased during the in vitro cocultivation when compared with the wild type. The phenotypes were co-cultivated with A. bisporus on compost with monitoring the green mould disease progression. The data indicated that prbi and ech42 genes is more important in disease progression than the p- 1,3 glucanase gene. Finally, the present study emphasises the role of the three cell wall degrading enzymes in green mould disease infection and may provide a promising tool for disease management.
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The biotransformation of water insoluble substrates by mammalian and bacterial cells has been problematic, since these whole cell reactions are primarily performed in an aqueous environment The implementation of a twophase or encapsulated system has the advantages of providing a low water system along with the physiological environment the cells require to sustain themselves. Encapsulation of mammalian cells by formation of polyamide capsules via interfacial polymerization illustrated that the cells could not survive this type of encapsulation process. Biotransformation of the steroid spironolactone [3] by human kidney carcinoma cells was performed in a substrate-encapsulated system, yielding canrenone [4] in 70% yield. Encapsulation of nitrile-metabolizing Rhodococcus rhodochrous cells using a polyamide membrane yielded leaky capsules, but biotransformation of 2-(4- chlorophenyl)-3-methylbutyronitrile (CPIN) [6] in a free cell system yielded CPIN amide [7] in 40% yield and 94% ee. A two-phase biotransformation of CPIN consisting of a 5:1 ratio of tris buffer, pH 7.2 to octane respectively, gave CPIN acid [8] in 30% yield and 97% ee. It was concluded that Rhodococcus rhodochrous ATCC 17895 contained a nonselective nitrile hydratase and a highly selective amidase enzyme.
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The first and rate-limiting step of lipolysis is the removal of the first fatty acid from a triglyceride molecule; it is catalyzed by adipose triglyceride lipase (ATGL). ATGL is co-activated by comparative gene identification-58 (CGI-58) and inhibited by the G(0)/G(1) switch gene-2 protein (G0S2). G0S2 has also recently been identified as a positive regulator of oxidative phosphorylation within the mitochondria. Previous research has demonstrated in cell culture, a dose dependent mechanism for inhibition by G0S2 on ATGL. However our data is not consistent with this hypothesis. There was no change in G0S2 protein content during an acute lipolytic inducing set of contractions in both whole muscle, and isolated mitochondria yet both ATGL and G0S2 increase following endurance training, in spite of the fact that there should be increased reliance on intramuscular lipolysis. Therefore, inhibition of ATGL by G0S2 appears to be regulated through more complicated intracellular or post-translation regulation.