Structural study on lipid A and the O-specific polysaccharide of the lipopolysaccharide from a clinical isolate of Bacteroides vulgatus from a patient with Crohn's disease.

Bacteroides vulgatus has been shown to be involved in the aggravation of colitis. Previously, we separated two potent virulence factors, capsular polysaccharide (CPS) and lipopolysaccharide (LPS), from a clinical isolate of B. vulgatus and characterized the structure of CPS. In this study, we elucidated the structures of O-antigen polysaccharide (OPS) and lipid A in the LPS. LPS was subjected to weak acid hydrolysis to produce the lipid A fraction and polysaccharide fraction. Lipid A was isolated by preparative TLC, and its structure determined by MS and NMR to be similar to that of Bacteroides fragilis except for the number of fatty acids. The polysaccharide fraction was subjected to gel-filtration chromatography to give an OPS-rich fraction. The structure of OPS was determined by chemical analysis and NMR spectroscopy to be a polysaccharide composed of the following repeating unit: [-->4)alpha-L-Rhap(1-->3)beta-D-Manp(1-->].     

A GREEN DINOFLAGELLATE WITH CHLOROPHYLLS a and b: MORPHOLOGY,FINE STRUCTURE OF THE CHLOROPLAST AND CHLOROPHYLL COMPOSITION1

A green-colored marine unicell has been grown in unialgal culture and its morphology, chloroplast fine structure, and chlorophyll composition investigated. The organism is typical of dinoflagellates in its shape, flagellation, nucleus, mitochondria, and trichocysts. It is similar to Gymnodinium but possesses fine body scales. Chloroplasts and two kinds of vesicles bounded by double membranes, but no organelles obviously identifiable as nuclei or mitochondria, are associated in ribosome-dense cytoplasm separated by a double membrane from the dinophycean cytoplasm. The chloroplasts are unlike any previously reported for dinoflagellates. Each is enclosed by an envelope consisting of a double membrane. Chloroplast lamellae consist of three appressed thylakoids. Interlamellar pyrenoids are present. Pigment analysis reveals chlorophylls a and b but not chlorophyll c. It seems likely that the organism is an undescribed dinoflagellate containing an endosymbiont with chlorophylls a and b and that the reduction of the endosymbiont nucleus and mitochondria has permitted a more initmate symbiosis.     

The 21-kDa Polypeptide (VAP21) in the Rabies Virion Is a CD99-Related Host Cell Protein

In our monoclonal antibody (MAb) stocks prepared against the BHK-21 cell antigens, two (#11875 and 28276) recognized a 21-kDa polypeptide (referred to as VAP21) which is efficiently incorporated into the rabies virion. By using these MAbs, we isolated the cDNA clones that encoded a polypeptide of 144 amino acids from our BHK-21 cell cDNA library. Based on the following evidence, the cDNA was assumed to encode a full-length sequence of VAP21 antigen: i) expression of the cDNA in animal cells resulted in the production of a polypeptide recognized by the two MAbs, and its electrophoretic mobility was the same as that of authentic VAP21 antigen; and ii) immunization with the products from the cDNA-transformed E. coli cells raised specific antibodies in rabbits that recognized a 21-kDa polypeptide in the virion. From the deduced amino acid sequence, it is suggested that the VAP21 antigen has a molecular structure of type-I transmembrane protein containing characteristic proline-rich and glycine-rich regions in its ectodomain. Homology searches resulted in finding homologous sequences (totally about 40% homology) in the human MIC2 gene product (CD99; 32-kDa) of T lymphocytes. These results suggest that the VAP21 antigen in the rabies virion is a cellular CD99-related transmembrane protein.     

Chromosome numbers of some North American Salix

Chromosome numbers are reported for 19 species of North AmericanSalix, one natural hybrid, and one introduced species. The following 17 species are here examined cytologically for the first time:Salix amygdaloides Anderss.,S. arbusculoides Anderss.,S. brachycarpa Nutt.,S. Candida Willd.,S. discolor Muhl.,S. exigua Nutt.,S. kumilis Marsh.,S. interior Rowlee,S. lutea Nutt.,S. maccalliana Rowlee,S. monticola Rydb.,S. myrtillifolia Anderss.,S. pellita Anderss.,S. petiolaris J. E. Sm,S. scouleriana Barr.,S. silicicola Raup, andS. subcoerulea Piper. Of the 19 native North American species 11 are diploid, four are tetraploid, one is triploid, one hexaploid, one dodecaploid, and one exhibits more than one ploidal level.     

Effects of genetic backgrounds on hyperbilirubinemia in radixin-deficient mice due to different expression levels of Mrp3

ERM (ezrin/radixin/moesin) proteins are organizers of apical actin cortical layer in general. We previously reported that the knockout of radixin resulted in Rdx(-/-) mice with displacement/loss of the canalicular transporter Mrp2, giving rise to Dubin-Johnson syndrome-like conjugated hyperbilirubinemia in the mixed genetic background (C57BL/6-129/Sv) (Kikuchi, et al. (2002) Nature Genetics 31, 320-325). However, when these mice were kept under mixed genetic background for years (late mixed backgrounds; LMB), the conjugated hyperbilirubinemia gradually became inconspicuous, while evidence of liver injury increased. We examined the effect of genetic background by backcrossing LMB Rdx(-/-) mice to C57BL/6 and 129/Sv wild type mice with the result that the Rdx(-/-) congenic mice regained hyperbilirubinemia with reduced hepatocellular damage. As revealed by immunofluorescence and western blots, the localization/expression of apical transporters, Mrp2, CD26, P-gps, and Bsep were not influenced by backcrossing, though those of a basolateral transporter, Mrp3, were strikingly increased by backcrossing.     

Glucose 1-phosphate increases active transport of calcium in intestine

Active calcium transport in intestine is essential for serum calcium homeostasis as well as for bone formation. It is well recognized that vitamin D is a major, if not sole, stimulator of intestinal calcium transport activity in mammals. Besides vitamin D, endogenous glucose 1-phosphate (G1P) affects calcium transport activity in some microorganisms. In this study, we investigated whether G1P affects intestinal calcium transport activity in mammals as well. Of several glycolytic intermediates, G1P was the sole sugar compound in stimulating intestinal calcium uptake in Caco-2 cells. G1P stimulated net calcium influx and expression of calbindin D9K protein in rat intestine, through an active transport mechanism. Calcium uptake in G1P-supplemented rats was greater than that in the control rats fed a diet containing adequate vitamin D3. Bone mineral density (BMD) of aged rat femoral metaphysis and diaphysis was also increased by feeding the G1P diet. G1P did not affect serum levels of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] at all. These results suggest that exogenously applied G1P stimulates active transport of calcium in intestine, independent of vitamin D, leading to an increase of BMD.     

Marine growth and survival of white-spotted charr,Salvelinus leucomaenis, in relation to smolt size

The relationships between marine growth and survival, and smolt sizes were examined for white-spotted charr (Salvelinus leucomaenis) populations in the Nairo River, Rebun Island, off northern Hokkaido Island, and the Haraki River, southern Hokkaido Island, Japan. Smolts in the Haraki River were much larger in size than those in the Nairo River, despite being similar size among age cohorts and between the sexes in each population. From scale analyses, smolt size-specific growth rates and survival were estimated by utilizing both observed and back-calculated size-frequencies. Inverse correlations between specific growth rates at sea and smolt size were found in both populations. However, the expected size increments at a given size of smolts in the Haraki River were significantly larger than of those in the Nairo River. For both populations, there were no significant effects of smolt ages (2 to 5 or 6 years) on growth rates at sea. Larger smolts had a consistent survival advantage, although the degree of size-dependent survival function seemed to differ between populations. The results suggested that growth and survival of individuals at sea are determined ultimately by size-, rather than age-, dependent factors depending upon local environmental conditions, supporting the hypothesis of a threshold size for smolting within populations.