Low-sulfated chondroitin sulfate in human blood and urine

Blood and urinary low-sulfated chondroitin sulfate from healthy young and aged volunteers have been characterized by gel chromatography, two-dimensional electrophoresis on cellulose acetate strips and by chemical and enzymatic analysis. No difference in content of the material (24 nmol hexosamine per ml plasma) was observed regardless of age. Chemical composition (approximately 40% sulfation at 4-position of galactosamine) and molecular weight (about 8000) of blood and urinary low-sulfated chondroitin sulfates were found to be the same, though urinary excretion of the material was much higher in the aged than in the young adults (Ohkawa et al. (1972) J. Biochem. 72, 1495–1501). Low-sulfated chondroitin sulfate in serum was in a bound form with a molecular weight of more than 100000, irrespective of age. These results suggest that increase in urinary excretion of low-sulfated chondroitin sulfate in the aged is mainly due to renal dysfunction.Low-sulfated chondroitin sulfate was also the main component of acidic glycosaminoglycans in blood from patients with Hurler's syndrome who excreted excessive amounts of dermatan sulfate and heparan sulfate in urine. This suggests that low-sulfated chondroitin sulfate in blood is not merely a precursor of urinary glycosaminoglycans in the case of healthy young adults.     

Drug Resistance of Enteric Bacteria

Drug resistance of 3,000 Shigella strains isolated in 1965 were investigated. These strains originated from 10 City Hospitals and 4 Prefectural Health Centers, which are located in different parts of Japan. One hundred and seventy strains which were resistant to 4 drugs, chloramphenicol (CM), tetracycline (TC), dihydrostreptomycin (SM), and sulfanilamide (SA), were selected at random from these stock cultures in this laboratory and the distribution of R factors in these isolates was examined. It was found that the strains all harbored R factors which were capable of transferring drug resistance by usual conjugal process. Among the strains carrying R factors, 85 per cent harbored a single type of R factor and 15 per cent carried two types of R factor in a cell. The latter is called the hetero-R state. Among the strains in the hetero-R state, isolation of strains harboring both R (SM.SA) and R (TC.CM.SM.SA) factors was most frequent. It was found that 25 R (SM.SA) factors isolated from strains in hetero-R had the genetic determinant iR^?, while most of the R (TC.CM.SM.SA) factors isolated from natural sources were iR⁺. When two types of R factor, R (SM.SA) and R (TC.CM.SM.SA) derived from the same host cells, were brought together in a host cell by superinfection with both factors, they were found to exist stably in a host bacterium. These results confirmed the stable existence of both factors in Shigella strains isolated from dysenteric patients.     

Development of an anti-hepatitis B virus (HBV) agent through the structure-activity relationship of the interferon-like small compound CDM-3008

Hepatitis B, a viral infectious disease caused by hepatitis B virus (HBV), is a life-threatening disease that leads liver cirrhosis and liver cancer. Because the current treatments for HBV, such as an interferon (IFN) formulation or nucleoside/nucleotide analogues, are not sufficient, the development of a more effective agent for HBV is urgent required.CDM-3008 (1, 2-(2,4-bis(trifluoromethyl)imidazo[1,2-a][1,8]naphthyridin-8-yl)-1,3,4-oxadiazole) (RO8191)) is a small molecule with an imidazo[1,2-a][1,8]naphthyridine scaffold that shows anti-HCV activity with an IFN-like effect. Here, we report that 1 was also effective for HBV, although the solubility and metabolic stability were insufficient for clinical use. Through the structure-activity relationship (SAR), we discovered that CDM-3032 (11, N-(piperidine-4-yl)-2,4-bis(trifluoromethyl)imidazo[1,2-a][1,8]naphthyridine-8-carboxamide hydrochloride) was more soluble than 1 (>30?mg/mL for 11 versus 0.92?mg/mL for 1). In addition, the half-life period of 11 was dramatically improved in both mouse and human hepatic microsomes (T1/2, >120?min versus 58.2?min in mouse, and >120?min versus 34.1?min in human, for 11 and 1, respectively).     

Vasomotor effects of noradrenaline, acetylcholine, histamine, 5-hydroxytryptamine and bradykinin on snake (Trimeresurus flavoviridis) basilar arteries

We investigated the responsiveness of basilar arterial rings isolated from snakes to noradrenaline (NA), acetylcholine (ACh), histamine (His), 5-hydroxytryptamine (5-HT), mammalian bradykinin (BK) and rattlesnake BK. We also examined whether endothelial cells were involved in the responsiveness to ACh, BK, rattlesnake BK and in their resting vascular tone. NA and 5-HT induced concentration-dependent contractions. The cumulative concentration response curves of NA and 5-HT were shifted to the right in parallel by phentolamine (an alpha antagonist) and methiothepin (a 5-HT(1) and 5-HT(2) antagonist), respectively. However, ketanserin (a 5-HT(2) antagonist) had no effect on the cumulative concentration response curve of 5-HT. His, ACh, BK and rattlesnake BK had no effect on resting vascular tone; however, rattlesnake BK and sodium nitroprusside relaxed arteries precontracted by 5-HT. The rattlesnake BK-induced relaxations were almost abolished by L-nitro arginine (L-NA, a nitric oxide synthase inhibitor). L-NA and indomethacin (a cyclooxygenase inhibitor) had no effect on resting vascular tone or on precontracted arteries. These results suggest that alpha and 5-HT(1) receptor subtypes might be important in arterial contraction. Endothelial cells might play an important role in the responsiveness of snake basilar arteries to rattlesnake BK, but they might not be involved in the responsiveness to ACh, BK and in resting vascular tone.     

Glycolipids with nonreducing end alpha-mannosyl residues that have the potential to activate invariant Valpha19 NKT cells

We have previously demonstrated that alpha-mannosyl ceramide and its derivatives promote immune responses of NK1.1(+) invariant Valpha19-Jalpha33 T cell receptor (TCR) alpha(+) T cells (Valpha19 NKT cells). In this study, attempts were made to determine the structural requirements for natural ligands for Valpha19 NKT cells. Naturally occurring and synthetic glycolipids were analyzed for their ability to stimulate the cells prepared from invariant Valpha19-Jalpha33 TCR transgenic mice, in which development of Valpha19 NKT cells is facilitated. As a result, alpha-mannosyl phosphatidylinositols such as 2,6-di-alpha-mannosyl phosphatidylinositol and alpha-mannosyl-4alpha-glucosaminyl-6-phosphatidylinositol (alpha-Man-GlcNH(2)-PtdIns) as well as alpha-mannosyl ceramide derivatives were found to activate the cells from the transgenic mouse liver, gut lamina propria and spleen in vivo and in vitro. Thus, glycolipids with nonreducing end alpha-mannosyl residues are suggested to be potent antigens for Valpha19 NKT cells. Next, a series of invariant Valpha19-Jalpha33 TCR(+) hybridomas, each with variations in the sequence of the Valpha-Jalpha junction and the TCR beta chain, were tested for responsiveness toward the alpha-mannosyl glycolipids. A loose correlation between the primary structure of the TCR and the reactive glycolipids was observed. For instance, hybridomas expressing TCRs consisting of an alpha chain with a variation in the Valpha19-Jalpha33 junction and a Vbeta6(+)beta chain showed affinity towards alpha-mannosyl ceramide and alpha-Man-GlcNH(2)-PtdIns, whereas those expressing TCRs with an invariant Valpha19-Jalpha33 alpha chain and a Vbeta8(+)beta chain responded to 2,6-di-alpha-mannosyl phosphatidylinositol. Thus, it is suggested that Valpha19 NKT cells with microheterogeneity in the TCR structure have been generated for defense against various antigens expressing alpha-mannosyl glycolipids.     

Note on <Emphasis Type="Italic">Cordyceps brongniartii</Emphasis> Shimazu collected from the wild in Japan

We collected Cordyceps brongniartii from the wild associated with coleopteran larvae for the first time in Japan. Morphological comparisons of C. brongniartii with the type specimen showed slight morphological difference, whereas it showed considerable differences from those collected in the wild in China. PCR-RFLP and ITS sequence analyses corroborated the teleomorph–anamorph relationship between C. brongniartii and Beauveria brongniartii.     

Two cases of young adults sleepwalking

Sleep and Biological Rhythms - We report two cases of young adults sleepwalking. The first was a 19-year-old man whose episode occurred at the end of the first sleep stage 4. The second patient, a...     

Comparative evaluation of gene-set analysis methods

Background  

Multiple data-analytic methods have been proposed for evaluating gene-expression levels in specific biological pathways, assessing differential expression associated with a binary phenotype. Following Goeman and Bühlmann's recent review, we compared statistical performance of three methods, namely Global Test, ANCOVA Global Test, and SAM-GS, that test "self-contained null hypotheses" Via. subject sampling. The three methods were compared based on a simulation experiment and analyses of three real-world microarray datasets.