Evaluation of Oriental medicines using a cultured renal epithelial cell line, LLC-PK_1 : Effects of Carthami Flos, Rhei Rhizoma and Astragali Radix

The effects of Carthami Flos，Rhei Rhizoma and Astragali Radix extract，which have been proved to ameliorate renal failure in rats with glycerol-induced renal failure，were examined in cell culture under three different culture conditions．Rhei Rhizoma　extract showed the most potent effect among the three crude drugs under any of the routine，hypoxia-reoxygenation and cisplatin exposure conditions employed，suggesting that its favorable effect on proximal tubule function is due to suppression　of lipid peroxidation via free radicals．In contrast，this anti-cytotoxic activity was low in Astragali Radix extract，and almost nil in Carthami Flos extract．These results indicate that Rhei Rhizoma，Astragali Radix and Carthami Flos exert their actions on different sites in the kidney

Molecular Basis and Consequences of the Cytochrome c-tRNA Interaction.

The intrinsic apoptosis pathway occurs through the release of mitochondrial cytochrome c to the cytosol, where it promotes activation of the caspase family of proteases. The observation that tRNA binds to cytochrome c revealed a previously unexpected mode of apoptotic regulation. However, the molecular characteristics of this interaction, and its impact on each interaction partner, are not well understood. Using a novel fluorescence assay, we show here that cytochrome c binds to tRNA with an affinity comparable with other tRNA-protein binding interactions and with a molecular ratio of ∼3:1. Cytochrome c recognizes the tertiary structural features of tRNA, particularly in the core region. This binding is independent of the charging state of tRNA but is regulated by the redox state of cytochrome c. Compared with reduced cytochrome c, oxidized cytochrome c binds to tRNA with a weaker affinity, which correlates with its stronger pro-apoptotic activity. tRNA binding both facilitates cytochrome c reduction and inhibits the peroxidase activity of cytochrome c, which is involved in its release from mitochondria. Together, these findings provide new insights into the cytochrome c-tRNA interaction and apoptotic regulation

Control of catalytic cycle by a pair of analogous tRNA modification enzymes.

Enzymes that use distinct active site structures to perform identical reactions are known as analogous enzymes. The isolation of analogous enzymes suggests the existence of multiple enzyme structural pathways that can catalyze the same chemical reaction. A fundamental question concerning analogous enzymes is whether their distinct active-site structures would confer the same or different kinetic constraints to the chemical reaction, particularly with respect to the control of enzyme turnover. Here, we address this question with the analogous enzymes of bacterial TrmD and its eukaryotic and archaeal counterpart Trm5. TrmD and Trm5 catalyze methyl transfer to synthesize the m1G37 base at the 3\u27 position adjacent to the tRNA anticodon, using S-adenosyl methionine (AdoMet) as the methyl donor. TrmD features a trefoil-knot active-site structure whereas Trm5 features the Rossmann fold. Pre-steady-state analysis revealed that product synthesis by TrmD proceeds linearly with time, whereas that by Trm5 exhibits a rapid burst followed by a slower and linear increase with time. The burst kinetics of Trm5 suggests that product release is the rate-limiting step of the catalytic cycle, consistent with the observation of higher enzyme affinity to the products of tRNA and AdoMet. In contrast, the lack of burst kinetics of TrmD suggests that its turnover is controlled by a step required for product synthesis. Although TrmD exists as a homodimer, it showed half-of-the-sites reactivity for tRNA binding and product synthesis. The kinetic differences between TrmD and Trm5 are parallel with those between the two classes of aminoacyl-tRNA synthetases, which use distinct active site structures to catalyze tRNA aminoacylation. This parallel suggests that the findings have a fundamental importance for enzymes that catalyze both methyl and aminoacyl transfer to tRNA in the decoding process

Potential for interdependent development of tRNA determinants for aminoacylation and ribosome decoding.

Although the nucleotides in tRNA required for aminoacylation are conserved in evolution, bacterial aminoacyl-transfer RNA synthetases are unable to acylate eukaryotic tRNA. The cross-species barrier may be due to the absence of eukaryote-specific domains from bacterial aminoacyl-transfer RNA synthetases. Here we show that whereas Escherichia coli CysRS cannot acylate human tRNA(Cys), the fusion of a eukaryote-specific domain of human CysRS overcomes the cross-species barrier in human tRNA(Cys). In addition to enabling recognition of the sequence differences in the tertiary core of tRNA(Cys), the fused eukaryotic domain redirects the specificity of E. coli CysRS from the A37 present in bacterial tRNA(Cys) to the G37 in mammals. Further experiments show that the accuracy of codon recognition on the ribosome was also highly sensitive to the A37G transition in tRNA(Cys). These results raise the possibility of the development of tRNA nucleotide determinants for aminoacylation being interdependent with those for ribosome decoding

A tunable plasmonic refractive index sensor with nanoring-strip graphene arrays

In this paper, a tunable plasmonic refractive index sensor with nanoring-strip graphene arrays is numerically investigated by the finite difference time domain (FDTD) method. The simulation results exhibit that by changing the sensing medium refractive index nmed of the structure, the sensing range of the system is large. By changing the doping level ng, we noticed that the transmission characteristics can be adjusted flexibly. The resonance wavelength remains entirely the same and the transmission dip enhancement over a big range of incidence angles [0,45] for both TM and TE polarizations, which indicates that the resonance of the graphene nanoring-strip arrays is insensitive to angle polarization. The above results are undoubtedly a new way to realize various tunable plasmon devices, and may have a great application prospect in biosensing, detection and imaging

Detection of hepatitis B virus DNA among accepted blood donors in Nanjing, China

<p>Abstract</p> <p>Background</p> <p>Posttransfusion hepatitis B virus (HBV) infection still occurs although its incidence has been substantially reduced since the introduction of screening of hepatitis B surface antigen (HBsAg) in blood donors. This study aimed to investigate the occult HBV infection in accepted blood donors in Nanjing, China.</p> <p>Results</p> <p>The lower detection limit of the nested PCR in this study was estimated to be 20 copies/ml HBV DNA. The positive rate of occult HBV infection was 0.13% (5 of 2972) in the accepted blood donors. Sequencing data showed that the amplified HBV sequences were not identical each other and to the known sequences cloned in our laboratory, excluding the false-positive caused by cross-contamination. Phylogenetic analysis showed that the HBV in all five donors was genotype B; a single base deletion was detected in the S region of HBV DNA from one donor, and no mutation was observed in the "a" determinant of HBsAg from four other donors. All five donors were negative for anti-HBs and one was positive for anti-HBc.</p> <p>Conclusions</p> <p>The prevalence of occult HBV infection in the accepted blood donors in Nanjing, China is relatively high. The data would be meaningful in adapting strategy to eliminate posttransfusion HBV infection in China.</p