Xaa-Arg-Gly triplets in the collagen triple helix are dominant binding sites for the molecular chaperone HSP47.

HSP47 is an essential procollagen-specific molecular chaperone that resides in the endoplasmic reticulum of procollagen-producing cells. Recent advances have revealed that HSP47 recognizes the (Pro-Pro-Gly)(n) sequence but not (Pro-Hyp-Gly)(n) and that HSP47 recognizes the triple-helical conformation. In this study, to better understand the substrate recognition by HSP47, we synthesized various collagen model peptides and examined their interaction with HSP47 in vitro.

Direct determination of interfacial magnetic moments with a magnetic phase transition in Co nanoclusters on Au(111).

The spin, in-plane and out-of-plane orbital and magnetic dipole moments of almost purely interfacial Co atoms were directly determined for Au/2-monolayer Co nanoclusters/Au(111) by angle-dependent magnetic circular x-ray dichroism (MCXD) measurements. The field- and temperature-dependent MCXD evidences a ferromagnetic(FM)-to-superparamagnetic phase transition in single-domain clusters with decreasing size. The interfacial moments are remarkably enhanced as compared with bulk values, verifying theoretical predictions.

Close correlation between the magnetic moments, lattice distortions, and hybridization in LaMnO3 and La(1-x)Sr(x)MnO3+delta: doping-dependent magnetic circular X-ray dichroism study.

The first observation of a magnetic circular x-ray dichroism (MCXD) at the Mn L2,3 core edges in antiferromagnetic LaMnO3 shows canted spin and orbital ( m(orb)) moments arising from lattice distortions. An L2,3-edge MCXD in ferromagnetic metals and insulators, La1-xSr(x)MnO3+delta, reveals that m(orb) of Mn strongly depends on x in the metallic regime but remains unchanged with the metal-to-insulator transition (x approximately 0.16).

New sesquiterpene lactones from Elephantopus mollis and their leishmanicidal activities.

The leishmanicidal compounds isolated from whole plants of Elephantopus mollis H.B.K.

[Estradiol Reference Standard (Control 001) of National Institute of Health Sciences].

The raw material of tocopherol was tested for the preparation of "Estradiol Reference Standard (Control 991)". Analytical data obtained were: melting point, 178.5 degrees C: UV spectrum, lambda max of 281 nm and specific absorbance in ethanol at lambda max = 77.

[Prednisolone Sodium Phosphate Reference Standard (Control 001) of National Institute of Health Sciences].

The raw material for prednisolone sodium phosphate was examined for the preparation of the "Prednisolone Sodium Phosphate Reference Standard (Control 001)". The analytical data obtained were: pH, 7.9; optical rotation, [alpha]D20 = +98.

[Tocopherol Reference Standard (Control 001) of National Institute of Health Sciences].

The raw material of tocopherol was tested for the preparation of "Tocopherol Reference Standard (Control 991)". Analytical data obtained were: IR spectrum, same as the Tocopherol Reference Standard (Control 941); specific absorbance, E/cm% (292 nm) = 72.9; thin-layer chromatography, no impurities were detected until 50.

[Tocopherol Acetate Reference Standard (Control 001) of National Institute of Health Sciences].

The raw material of tocopherol acetate was examined for the preparation of the "Tocopherol Acetate Reference Standard (Control 001)". Analytical data obtained were: IR spectrum, same as that of the Tocopherol Acetate Reference Standard (Control 974); specific absorbance, E/cm% (284 nm) = 43.7; thin-layer chromatography, no impurities were detected until 50 micrograms of the loaded raw material; high-performance liquid chromatography (HPLC), total amount of impurities estimated to be less than 0.

[Thiamine Hydrochloride Solution Reference Standard (Control 991) of National Institute of Health Sciences].

The raw material of thiamine hydrochloride solution was examined for preparation of the "Thiamine Hydrochloride Solution Reference Standard (Control 991)". The analytical data obtained were: assay by HPLC, 101.0%; spectrophotometric assay, 100.

Active repression of RAR signaling is required for head formation.

The retinoic acid receptors (RARs) recruit coactivator and corepressor proteins to activate or repress the transcription of target genes depending on the presence of retinoic acid (RA). Despite a detailed molecular understanding of how corepressor complexes function, there is no in vivo evidence to support a necessary function for RAR-mediated repression. Signaling through RARs is required for patterning along the anteroposterior (A-P) axis, particularly in the hindbrain and posterior, although the absence of RA is required for correct anterior patterning.

Mechanistic study of enantiomeric recognition of primary amino compounds using an achiral crown ether with cyclodextrin by capillary electrophoresis and nuclear magnetic resonance.

A model and theoretical equations are presented to investigate the enantiomeric recognition mechanism of primary amino compounds using an achiral crown ether with cyclodextrin by capillary electrophoresis (CE) and nuclear magnetic resonance (NMR). Association constants were calculated from CE and 1H NMR experiment results on the basis of the model and the equations. The key step of chiral recognition was identified from those values.

[Prophylactic effect of pirarubicin (THP) on postoperative recurrence of superficial bladder cancer in terms of intravesical retention time].

In order to determine the modality of prophylactic intravesical instillation of pirarubicin (THP = tetrahydropyranyladriamycin) following transurethral resection (TUR) of superficial bladder cancer, a prospective randomized study was performed. A total of 79 patients were randomized into "2-hour instillation" (A), "5-min instillation" (B) and "control" (C) groups. Prophylactic efficacy and side effects were analyzed in each group.

A rescue factor abolishing neuronal cell death by a wide spectrum of familial Alzheimer's disease genes and Abeta.

Through functional expression screening, we identified a gene, designated Humanin (HN) cDNA, which encodes a short polypeptide and abolishes death of neuronal cells caused by multiple different types of familial Alzheimer's disease genes and by Abeta amyloid, without effect on death by Q79 or superoxide dismutase-1 mutants. Transfected HN cDNA was transcribed to the corresponding polypeptide and then was secreted into the cultured medium. The rescue action clearly depended on the primary structure of HN.

Enantiomeric separations of primary amino compounds by capillary electrochromatography with monolithic chiral stationary phases of chiral crown ether-bonded negatively charged polyacrylamide gels.

A novel enantiomeric separation method by capillary electrochromatography with chiral crown ether-bonded negatively charged polyacrylamide gels is presented. Two kinds of chiral crown ether derivatives, (+)-tetraallyl 18-crown-6 carboxylate and (+)-18-crown-6 tetracarboxylic acid 2-allyl ester were synthesized and allowed to covalently bind to a negatively charged polyacrylamide gel, a so-called monolithic stationary phase, respectively. The gel was placed in fused-silica tubing, the walls of which had been activated with a bifunctional reagent to make the resulting gel bind covalently to the inner surface.

Stimulation of corticospinal tract regeneration in the chronically injured spinal cord.

Acute spinal cord injury models have proved popular in studies aimed at identifying factors capable of influencing axonal regeneration within the central nervous system. In these models, the test factors (e.g.

Neuroprotective effect and intraocular penetration of nipradilol, a beta-blocker with nitric oxide donative action.

Purpose: To investigate the effect of nipradilol, an alpha(1),beta-blocker with a nitric oxide donative action, on N:-methyl-D-aspartate (NMDA)-induced retinal damage in rats and to determine whether topically instilled nipradilol penetrates the ipsilateral posterior retina-choroid at pharmacologically active concentrations in rabbits.

Methods: To determine effects on NMDA-induced damage, drugs were injected alone or with NMDA into the vitreous of one eye, and cell loss in the ganglion cell layer (GCL) and thinning of the retinal neural cell layers were histologically evaluated. To evaluate posterior penetration, first, [(14)C]-nipradilol was instilled, and its tissue concentration was measured.

Preparation of anhydrothrombin and characterization of its interaction with natural thrombin substrates.

Thrombin is a serine proteinase that plays a key role in thrombosis and haemostasis through its interaction with several coagulation factors. Anhydrothrombin was prepared from PMSF-inactivated thrombin under alkaline conditions, and the folded anhydrothrombin was successfully recovered after dialysis in the presence of glycerol. Anhydro-derivatives of factor Xa, factor VIIa and activated protein C could also be prepared essentially by the same procedure.

Molecular cloning of a novel ubiquitin-like protein, UBIN, that binds to ER targeting signal sequences.

To identify proteins that interact with HSP47, an endoplasmic reticulum (ER)-resident molecular chaperone, a yeast two-hybrid screening was performed using mouse full-length HSP47 including an N-terminal signal sequence as a bait. Analysis of several positive clones led to the identification and cloning of a novel gene, ubin, encoding a ubiquitin-like protein. Unlike other ubiquitin-like proteins, UBIN was shown to interact with signal sequences of various secretory and ER-luminal proteins, including HSP47, but not interact with signal sequences of mitochondrial targeting in two-hybrid system.

Substrate recognition by the collagen-binding domain of Clostridium histolyticum class I collagenase.

Clostridium histolyticum type I collagenase (ColG) has a segmental structure, S1+S2+S3a+S3b. S3a and S3b bound to insoluble collagen, but S2 did not, thus indicating that S3 forms a collagen-binding domain (CBD). Because S3a+S3b showed the most efficient binding to substrate, cooperative binding by both domains was suggested for the enzyme.

Differential chromatin packaging of genomic imprinted regions between expressed and non-expressed alleles.

Chromosomal regions subject to genomic imprinting comprise a functional domain exhibiting parental-specific expression of genes and hence may take a unique chromatin structure. Here we have examined the chromatin packaging state of allelic sites in the Zfp127/Snrpn locus on mouse chromosome 7 and in the Igf2r locus on mouse chromosome 17 with an assay consisting of chromatin fractionation and allele-specific detection. The results showed that non-transcribed alleles of Igf2r are packaged more compactly than transcribed alleles in F(1) hybrid mice of both types of cross between C57BL/6 and MSM strains, whereas a non-imprinted gene, Sod-2, in the vicinity of Igf2r does not show such a difference.

Histidine-rich glycoprotein (HRG) Tokushima 2: novel HRG deficiency, molecular and cellular characterization.

The proband, a 76-year-old woman, suffered from dural arteriovenous fistula. Her plasma histidine-rich glycoprotein (HRG) level was 50% of the normal level. A low level of plasma HRG was also found in her third daughter.

Enantiomeric separations of cationic and neutral compounds by capillary electrochromatography with monolithic chiral stationary phases of beta-cyclodextrin-bonded negatively charged polyacrylamide gels.

Enantiomeric separation by capillary electrochromatography with beta-cyclodextrin-bonded negatively charged polyacrylamide gels was examined. The columns used are capillaries filled with a negatively charged polyacrylamide gel, a so-called monolithic stationary phase, to which allyl carbamoylated beta-CD (AC-beta-CD) derivatives covalently bind. The capillary wall is activated first with a bifunctional reagent to make the resulting gel bind covalently to the inner surface of the fused-silica tubing.

Endoplasmic reticulum (ER)-associated degradation of misfolded N-linked glycoproteins is suppressed upon inhibition of ER mannosidase I.

To examine the role of early carbohydrate recognition/trimming reactions in targeting endoplasmic reticulum (ER)-retained, misfolded glycoproteins for ER-associated degradation (ERAD), we have stably expressed the cog thyroglobulin (Tg) mutant cDNA in Chinese hamster ovary cells. We found that inhibitors of ER mannosidase I (but not other glycosidases) acutely suppressed Cog Tg degradation and also perturbed the ERAD process for Tg reduced with dithiothreitol as well as for gamma-carboxylation-deficient protein C expressed in warfarin-treated baby hamster kidney cells. Kifunensine inhibition of ER mannosidase I also suppressed ERAD in castanospermine-treated cells; thus, suppression of ERAD does not require lectin-like binding of ER chaperones calnexin and calreticulin to monoglucosylated oligosaccharides.

Secretion, gamma-carboxylation, and endoplasmic reticulum-associated degradation of chimeras with mutually exchanged Gla domain between human protein C and prothrombin.

Warfarin, an antagonist of vitamin K, causes diminution of vitamin K-dependent coagulation factors in the circulation. Although all vitamin K-dependent factors have Gla domains, the warfarin-induced decrease in their plasma concentration differs among factors. In warfarin-treated HepG2 cells, we found modest and severe intracellular degradation of prothrombin and protein C, respectively.