Split tolerance in nude mice transplanted with 2'-deoxyguanosine-treated allogeneic thymus lobes

To elucidate the acquisition of self tolerance in the thymus, full-allogeneic thymic chimeras were constructed. Athymic C3H and BALB/c nude mice were reconstituted with the thymic lobes of BALB/c and B10.BR fetuses, respectively, that were organ cultured for 5 days in the presence of 2'-deoxyguanosine. T cells in these chimeras were tolerized to the host MHC in both MLR and CTL assays. In contrast, T cells in the chimeras exhibited split tolerance for the thymic MHC haplotype. CTL specific for class I MHC of the thymic haplotype were generated not only from the peripheral T cells of the chimeras but also from thymocytes re-populated in the engrafted thymic lobes. However, T cells in these chimeras responded poorly to the class II MHC of the thymic haplotype in a standard MLR assay. In a syngeneic MLR culture upon stimulation with enriched APC of the thymic haplotype, only 22 to 48% of the responses were mediated by CD4+ cells, and proliferations of CD4- cells were prominent. There were no haplotype-specific suppressor cells detected which would cause the unresponsiveness to the thymic class II MHC. These results indicated that the thymic lobes treated with 2'-deoxyguanosine were defective in the ability to induce the transplantation tolerance for the class I MHC expressed on the thymus, although the same thymic lobes were able to induce the transplantation tolerance for the thymic class II MHC.     

Intracellular localization of histone deacetylase HDA6 in plants

Journal of Plant Research - Histone modification is an important epigenetic mechanism in eukaryotes. Histone acetyltransferase and deacetylase regulate histone acetylation levels antagonistically,...     

Duplexed, bead-based competitive assay for inhibitors of protein kinases.

BACKGROUND: Many cellular signal transduction cascades have protein kinases as critical components. Small molecule protein kinase inhibitors can be effective as laboratory probes and drugs. Methods that allow two or more kinases to be evaluated simultaneously for inhibition by a small molecule would allow unequivocal tests of specificity and selectivity of action of the small molecule. METHODS: Two hexahistidine-tagged activin receptor-like kinases were expressed in E. coli, purified, and bound to nickel beads. A fluorescent kinase ligand (F-KL) that binds to the ATP-binding site of these kinases with nanomolar affinity was developed. Binding of F-KL with kinase on the bead made the beads bright, and inhibitors decreased the brightness. RESULTS: A test panel of 17 nonfluorescent kinase inhibitors, spanning two orders of magnitude affinity for the kinases, gave K(d) values for the kinases that correlated well with a fluorescence polarization assay. Results were obtained for the kinases in duplex, using an autosampler to send beads from a 96-well plate to a flow cytometer in a format suitable for high throughput screening. CONCLUSIONS: Inhibitors of kinases can be measured in duplex in a high throughput format by flow cytometry, if a suitable fluorescent ligand is available.     

Erratum to: De novo sequencing and comparative analysis of expressed sequence tags from gynodioecious fig (Ficus carica L.) fruits: caprifig and common fig

Tree Genetics & Genomes - Negative correlation caused by competition among individuals and positive spatial correlation due to environmental heterogeneity may lead to biases in estimating...     

Electrochemical assay of the nitrate and nitrite reductase activities of Rhizobium japonicum

This work describes an electrochemical method for the determination of the nitrate and nitrite reductase activities of Rhizobium japonicum. The advantage of the method lies in the use of whole cells for the analysis and we earlier developed this protocol for the assay of NO. The results obtained are comparable to the spectrophotometric Griess assay. As the method is based on electrochemical reduction, the commonly interfering biological components like ascorbic acid, uric acid, dopamine, etc., will not interfere with the analysis. This method can be extended to the fabrication of biosensors for nitrate and nitrite using the same principle.     

A phenomics approach detected differential epigenetic growth regulation between inbreds and their hybrid in <Emphasis Type="Italic">Oryza sativa</Emphasis>

Epigenetic changes in chromatin can be induced upon hybridization, but their contribution to phenotypic changes in F1 hybrids is not known. In this study, we examined the effect of genome-wide alteration of epigenetic chromatin status on the growth of inbreds and their F1 hybrid by large-scale fine-time-lapse growth monitoring of rice (Oryza sativa L.) under a controlled environment. This allowed us to dissect seedling growth of inbreds and their hybrid into separate growth parameters, and to analyze the effect of disturbance of inert chromatin states on these parameters. We found that superior growth parameters are generally more sensitive to global inhibition of the activities of histone deacetylases (HDACs), but this higher sensitivity is not directly correlated to heterosis in F1. Unexpectedly, however, the rate of the initial exponential growth of shoots is sensitive to HDAC inhibition only in inbreds, but not in F1, irrespective of phenotypic superiority. Our phenomics approach has detected an inbred-specific dependence of basic growth on the inert state of chromatin that is lost in F1 hybrids.     

Novel emission properties of melem caused by the heavy metal effect of lanthanides(III) in a LB film.

A novel emissive molecular system is constructed by the intercalation of the fluorophore melem (triamino-tri-s-triazine) within a Langmuir-Blodgett (LB) film of stearic acid with the periodic arrangement of lanthanides (Ln(III)), mainly Pr(III) with supporting of Eu(III). From emission spectra, decay curves, quantum yields and XPS measurements, it is clarified that the external heavy metal effect of Pr(III) on melem is much stronger in the film than in the bulk solid state, resulting in producing an unusual triplet state of melem. The triplet state of melem in the LB film donates the excitation energy to Pr(III) in the LB film, which is completely different from the energy transfer pathway of Pr-melem complex in the solid state through the singlet state of melem.