Fisheries, management and conservation of the whale shark Rhincodon typus in Taiwan

The Taiwanese government introduced a catch-and-report system for the whale shark Rhincodon typus in 2001, and in the following year limited the total allowable catch (TAC) to 80 individuals. This limit was reduced over the following years, reaching a low of 30 individuals in 2007. In November 2007, a comprehensive ban on R. typus fishing came into effect and protection measures were adopted for the species. This study measured and recorded the total length (L(T) ), body mass and sex of 810 R. typus, using fisheries and other released data for the period 1995-2008. The mean L(T) of individuals caught from June to October was smaller than that for individuals caught from November to May. Mean annual catches in set nets were higher in south-western Taiwan than in other regions of Taiwan. The sex ratio showed that males were caught more frequently than females in Taiwanese waters, indicating sex segregation. For the first time, the stock structure, conservation and management regulation of R. typus in the north-west Pacific Ocean have been reviewed. The results provide a useful resource for future assessment of the possible development of R. typus ecotourism in Taiwan.     

Anti-inflammatory sesquiterpene lactones from the flower of Vernonia cinerea

Bioassay-guided fractionation of the hexane extract from the flowers of Vernonia cinerea (Asteraceae) led to the isolation of a new sesquiterpene lactone, 8α-hydroxyhirsutinolide (2), and a new naturally occurring derivative, 8α-hydroxyl-1-O-methylhirsutinolide (3), along with seven known compounds (1 and 4-9). The structures of the new compounds were determined by 1D and 2D NMR experiments and by comparison with the structure of compound 1, whose relative stereochemistry was determined by X-ray analysis. The isolated compounds were evaluated for their cancer chemopreventive potential based on their ability to inhibit nitric oxide (NO) production and tumor necrosis factor alpha (TNF-α)-induced NF-κB activity. Compounds 1, 2, 4, 5, and 9 inhibited TNF-α-induced NF-κB activity with IC(50) values of 3.1, 1.9, 0.6, 5.2, and 1.6μM, respectively; compounds 4 and 6-9 exhibited significant NO inhibitory activity with IC(50) values of 2.0, 1.5, 1.2, 2.7, and 2.4μM, respectively.     

Akt phosphorylation is essential for nuclear translocation and retention in NGF-stimulated PC12 cells

Nerve growth factor (NGF) elicits Akt translocation into the nucleus, where it phosphorylates nuclear targets. Here, we describe that Akt phosphorylation can promote the nuclear translocation of Akt and is necessary for its nuclear retention. Overexpression of Akt-K179A, T308A, S473A-mutant failed to show either nuclear translocation or nuclear Akt phosphorylation, whereas expression of wild-type counterpart elicited profound Akt phosphorylation and induced nuclear translocation under NGF stimulation. Employing the PI3K inhibitor and a variety of mutants PI3K, we showed that nuclear translocation of Akt was mediated by activation of PI3K, and Akt phosphorylation status in the nucleus required PI3K activity. Thus the activity of PI3K might contribute to the nuclear translocation of Akt, and that Akt phosphorylation is essential for its nuclear retention under NGF stimulation conditions.     

An affinity-based scoring scheme for predicting DNA-binding activities of modularly assembled zinc-finger proteins

Zinc-finger proteins (ZFPs) have long been recognized for their potential to manipulate genetic information because they can be engineered to bind novel DNA targets. Individual zinc-finger domains (ZFDs) bind specific DNA triplet sequences; their apparent modularity has led some groups to propose methods that allow virtually any desired DNA motif to be targeted in vitro. In practice, however, ZFPs engineered using this ‘modular assembly’ approach do not always function well in vivo. Here we report a modular assembly scoring strategy that both identifies combinations of modules least likely to function efficiently in vivo and provides accurate estimates of their relative binding affinities in vitro. Predicted binding affinities for 53 ‘three-finger’ ZFPs, computed based on energy contributions of the constituent modules, were highly correlated (r = 0.80) with activity levels measured in bacterial two-hybrid assays. Moreover, K_d values for seven modularly assembled ZFPs and their intended targets, measured using fluorescence anisotropy, were also highly correlated with predictions (r = 0.91). We propose that success rates for ZFP modular assembly can be significantly improved by exploiting the score-based strategy described here.     

Tyrosinase inhibitory effects of 1,3-diphenylpropanes from Broussonetia kazinoki

Six 1,3-diphenylpropanes exhibiting inhibitory activities against both the monophenolase and diphenolase actions of tyrosinase were isolated from the methanol (95%) extract of Broussonetia kazinoki. These compounds, 1–6, were identified as kazinol C (1), D (2), F (3), broussonin C (4), kazinol S (5) and kazinol T (6). The latter two species (5 and 6) emerged to be new 1,3-diphenylpropanes which we fully spectroscopically characterized. The IC₅₀ values of compounds (1, 3–5) for monophenolase inhibition were determined to range between 0.43 and 17.9 μM. Compounds 1 and 3–5 also inhibited diphenolase significantly with IC₅₀ values of 22.8, 1.7, 0.57, and 26.9 μM, respectively. All four active tyrosinase inhibitors (1, 3–5) were competitive inhibitors. Interestigly they all mainfested simple reversible slow-binding inhibition against diphenolase. The most potent inhibitor, compound 4 diplayed the following kinetic parameters k₃ = 0.0993 μM^?1 min^?1, k₄ = 0.0048 min^-1, and K_i^app = 0.0485 μM.     

Long-term gus expression from Gladiolus callus lines containing either a bar-uidA fusion gene or bar and uidA delivered on separate plasmids

Gus expression was determined for 19 lines of embryogenic Gladiolus callus that contained the 35S-bar-uidA-nos fusion gene and for 21 callus lines that had been cobombarded with the 35S-bar-nos and 35S-uidA-nos plasmid DNAs. These lines were selected for analysis because they grew vigorously on Murashige and Skoog’s medium supplemented with 6 mg l⁻¹ phosphinothricin. All 19 lines that contained the 35S-bar-uidA-nos fusion gene expressed gus compared to only 15 (71%) of the lines that had been cobombarded as determined by enzyme assay. The level of gus expression was significantly higher the first year for 12 callus lines containing the bar-uidA fusion gene as compared to 2 years later in culture. Southern hybridization confirmed integration of the uidA gene in all callus lines that had been bombarded with the 35S-bar-uidA-nos fusion gene. Two of the callus lines that had been cobombarded lacked the uidA gene, and another cobombarded line that did not express gus contained a truncated uidA gene. Two callus lines resulting from cobombardment showed gus expression in only a few cells indicating that gus expression was not completely silenced in these lines. Gus expression could not be reversed using 5-azacytidine in these two low-expressing lines, and Southern hybridization supported that methylation of the genomic DNA had not occurred. Average levels of gus expression were significantly higher, 8.9× , in cells with the 35S-bar-uidA-nos fusion gene compared to the cobombarded callus lines indicating the advantage of using a bar-uidA fusion gene for obtaining higher levels of gus expression in Gladiolus.