Cathepsin K-dependent toll-like receptor 9 signaling revealed in experimental arthritis

Cathepsin K was originally identified as an osteoclast-specific lysosomal protease, the inhibitor of which has been considered might have therapeutic potential. We show that inhibition of cathepsin K could potently suppress autoimmune inflammation of the joints as well as osteoclastic bone resorption in autoimmune arthritis. Furthermore, cathepsin K-/- mice were resistant to experimental autoimmune encephalomyelitis. Pharmacological inhibition or targeted disruption of cathepsin K resulted in defective Toll-like receptor 9 signaling in dendritic cells in response to unmethylated CpG DNA, which in turn led to attenuated induction of T helper 17 cells, without affecting the antigen-presenting ability of dendritic cells. These results suggest that cathepsin K plays an important role in the immune system and may serve as a valid therapeutic target in autoimmune diseases.     

Role for piRNAs and noncoding RNA in de novo DNA methylation of the imprinted mouse Rasgrf1 locus

Genomic imprinting causes parental origin-specific monoallelic gene expression through differential DNA methylation established in the parental germ line. However, the mechanisms underlying how specific sequences are selectively methylated are not fully understood. We have found that the components of the PIWI-interacting RNA (piRNA) pathway are required for de novo methylation of the differentially methylated region (DMR) of the imprinted mouse Rasgrf1 locus, but not other paternally imprinted loci. A retrotransposon sequence within a noncoding RNA spanning the DMR was targeted by piRNAs generated from a different locus. A direct repeat in the DMR, which is required for the methylation and imprinting of Rasgrf1, served as a promoter for this RNA. We propose a model in which piRNAs and a target RNA direct the sequence-specific methylation of Rasgrf1.     

General acid-base catalysis in the mechanism of a hepatitis delta virus ribozyme

Many protein enzymes use general acid-base catalysis as a way to increase reaction rates. The amino acid histidine is optimized for this function because it has a pK(a) (where K(a) is the acid dissociation constant) near physiological pH. The RNA enzyme (ribozyme) from hepatitis delta virus catalyzes self-cleavage of a phosphodiester bond. Reactivity-pH profiles in monovalent or divalent cations, as well as distance to the leaving-group oxygen, implicate cytosine 75 (C75) of the ribozyme as the general acid and ribozyme-bound hydrated metal hydroxide as the general base in the self-cleavage reaction. Moreover, C75 has a pK(a) perturbed to neutrality, making it "histidine-like." Anticooperative interaction is observed between protonated C75 and a metal ion, which serves to modulate the pK(a) of C75. General acid-base catalysis expands the catalytic repertoire of RNA and may provide improved rate acceleration.     

A planar rhombic charge-separated tetrasilacyclobutadiene

The cyclobutadiene (CBD) molecule C(4)H(4) deviates from a high-symmetry square geometry to compensate for its antiaromatic electronic structure. Here, we report a CBD silicon analog, Si(4)(EMind)(4) (1), stabilized by the bulky 1,1,7,7-tetraethyl-3,3,5,5-tetramethyl-s-hydrindacen-4-yl (EMind) groups, obtained as air- and moisture-sensitive orange crystals by the reduction of (EMind)SiBr(3) with three equivalents of lithium naphthalenide. X-ray crystallography reveals a planar and rhombic structure of the Si(4) four-membered ring, with alternating pyramidal and planar configurations at the silicon atoms. The large (29)Si chemical shift differences (Δδ > 350 parts per million) in the solid-state nuclear magnetic resonance spectra suggest a contribution of an alternately charge-separated structure. The rhombic-shaped charge-separated singlet state of compound 1 thus stabilizes its cyclic 4π-electron antiaromaticity in a manner that contrasts sharply with the bond-length alternation, characterizing the rectangular distortion of carbon-based CBD.     

Growth characteristic and sink strength of fruit at different CO2 concentrations in a Japanese and a Dutch tomato cultivar

The aim of the present study was to elucidate how fruit growth was limited by the source and sink capacities in a Japanese (‘Momotaro York’) and a Dutch (‘Dundee’) tomato cultivar. The two cultivars were grown hydroponically with a high-wire system in greenhouses for 25 weeks, and the growth characteristics and sink strength of fruit were determined. Fruits were pruned to four (4F) or one (1F) per truss. The latter were used to determine potential fruit growth, an indicator of fruit sink strength. Growth was also determined under normal (LC) and enriched (HC, 700 μmol mol⁻¹) CO₂ concentrations to examine the effect of source enhancement on fruit production. In both cultivars under normal CO₂, the growth rate of fruit pruned to 4F per truss was lower than that in 1F, indicating that maximum potential fruit growth was not achieved. Under HC conditions, fruit growth rate of ‘Dundee’ achieved in 4F trusses was lower than that in 1F. In ‘Momotaro York’ in HC, fruit growth in 4F trusses was close to potential. This implies that fruit growth was source-limited irrespective of CO₂ concentrations in ‘Dundee’ cultivar while fruit growth in ‘Momotaro York’ under normal and enriched CO₂ conditions was limited by source and sink strengths, respectively. Adjustments of cultural practices including increasing fruit number per truss and/or genetic approaches to enhancing fruit sink strength by breeding may improve fruit yields of Japanese cultivars under high source/sink conditions.     

Characterization of corrinoid compounds from a Japanese black tea (Batabata-cha) fermented by bacteria

A Japanese fermented black tea (Batabata-cha) contained a considerable amount of vitamin B(12) (456 +/- 39 ng per 100 g dry tea leaves and 2.0 +/- 0.3 ng per 100 mL of tea drink). A corrinoid compound was partially purified and characterized from the tea leaves. The patterns of the purified compound by the silica gel 60 thin-layer chromatography and C18 reversed phased high-performance liquid chromatography were identical to those of authentic vitamin B(12). When 20 week old vitamin B(12) deficient rats, which excreted substantial amounts (about 250 mg/day) of methylmalonic acid in urine as an index of vitamin B(12) deficiency, were fed the tea drink (50 mL/day, 1 ng of vitamin B(12)) for 6 weeks, urinary methylmalonic acid excretion (169 +/- 29 mg/day) of the tea drink-supplemented 26 week old rats decreased significantly relative to that (250 +/- 32 mg/day) of the deficient rats. The results indicate that the vitamin B(12) found in the fermented black tea is bioavailable in mammals.     

Effect of the number of intercrosses on grain yield in basic rice synthetic populations

The effect of 0, 1, 2, 3 and 4 intercross cycles using genetic male sterility on the mean yield and genetic variability of the CNA 5 irrigated rice synthetic population was assessed studying 300 S_0:2 (1998/99) and S_0:3 (1999/00) families grown in two locations (Lambari, MG and Goianira, GO). Each intercross was investigated using a 8 × 8 triple lattice design experiment containing 60 families and four control cultivars. Five experiments were carried out per location and generation, totaling 20 experiments. Individual, combined over locations, combined over generations and combined over locations and generations analyses of variance were performed. Linear regression equations were fitted to data of each location and generation as well as, genetic and phenotypic parameters were estimated. An average increase of 3.91% in grain yield was obtained with the increase in the number of intercrosses. However, when genetic gains obtained from selection of the five and ten highest yielding families were considered, negative b estimates were always obtained, suggesting a tendency of gain increase reduction with the increase in the number of intercrosses. This was confirmed by the analysis of the means of the best families, since increases in the number of intercrosses did not result in selection of families with greater mean grain yield. The estimates of the genetic variances and heritabilities did not suggest an increase in genetic variability with the increase in intercross cycles. Therefore, carrying out intercrosses on the base population did not improve the results of the irrigated rice recurrent selection program. This revised version was published online in August 2006 with corrections to the Cover Date.     

Morphology and water and lipid contents of stifle menisci of growing swine.

Morphology and distribution of water and lipid were examined in menisci of growing swine. The broadness of menisci and the size of protruded tissue in the inner anterior portion of menisci varied among animals. Small tears of tissues were frequently observed. Both the thickness and tissue weight were greater in the lateral than in the medial meniscus. Water content and intensity of glycosaminoglycan staining with safranin-O were found to be greatest in the inner one-third of the meniscus, and greater in the middle than in the outer one-third of the meniscus. Lipid content was greater in the outer one-third than in the inner two-thirds of the meniscus.