Monomeric, monovalent derivative of Maackia amurensis leukoagglutinin. Preparation and application to the study of cell surface glycoconjugates by flow cytometry

A stable subunit of Maackia amurensis leukoagglutinin (MMAL) was prepared by the selective reduction of disulfide bridges between the subunits followed by alkylation with 4-vinylpyridine. MMAL failed to precipitate fetuin, whereas it retained its ability to bind to the same glycoprotein coated on a plastic plate, indicating the monovalency of this derivative. This binding to immobilized fetuin was inhibited by a haptenic sugar, Neu5Ac alpha 2-3lactose, with the same inhibitory potency as against the native M. amurensis leukoagglutinin. Microscopic observation as well as flow cytometric analyses showed that Chinese hamster ovary cells were clearly stained with fluorescein isothiocyanate-labeled MMAL without any detectable agglutination. This staining was inhibited by the addition of fetuin or by the sugar chains of fetuin. Differences in the types of sialylated glycoconjugates on the cell surface of several cell lines were detected by the combined use of fluorescein isothiocyanate-labeled MMAL and the monomeric derivative of elderberry bark lectin (specific for the Neu5Ac alpha 2-6Gal/GalNAc sequence) by flow cytometry. These results demonstrate the usefulness of these monovalent derivatives of sialylated oligosaccharide-specific lectins as probes for the analysis of cell surface glycoconjugates containing sialic acid by the technique of flow cytometry.     

Characteristics of a continuous Si-Ti-C-O fibre with low oxygen content using an organometallic polymer precursor

A continuous Si-Ti-C-O fibre with 12 wt% oxygen content, which is lower than the usual 18 wt% found in the normal fibres, was synthesized by using polytitanocarbosilane which has fewer Si-Si bonds than the usual precursor polymer. The density, tensile strength, tensile modulus and thermal conductivity were found to be 2.37 g cm^–3, 3.4±0.3 GPa, 190±10 GPa and 1.40 W m^–1 K^–1, respectively. Amongst these properties, the tensile modulus was improved by 20 GPa and the thermal conductivity had a higher value in comparison with that of the ordinary Si-Ti-C-O fibre with 18 wt% oxygen content. The Si-Ti-C-O fibre with a 12 wt% oxygen content has a better heat resistance above 1400 °C in an argon atmosphere and 1300 °C in air, than the usual fibre. About 60 and 40% of its tensile strength at room temperature were retained in air at respectively, 1500 and 1600 °C. This improved ceramic fibre is considered to be useful as a reinforcing material for advanced composites such as high-temperature ceramic matrix composites and metal matrix composites.     

A monocyte chemotactic factor, S19 ribosomal protein dimer, in phagocytic clearance of apoptotic cells

HL-60 cells derived from a human promyelocytic leukemia underwent apoptosis by heat treatment. When the heat-treated HL-60 cells were injected into guinea pig skin, monocyte/macrophage infiltration was observed 24 or 36 hours later, and the apoptotic cells were phagocytically cleared by 48 hours after their injection. The infiltration and clearance patterns were quite different from those observed in injection of necrotic or boil-fixed HL-60 cells. The apoptotic cells released a monocyte chemotactic factor in vitro 24 hours after the heat treatment. The chemotactic factor generated was identified as the cross-linked homodimer of S19 ribosomal protein by its immunologic and physicochemical properties. A serine protease that inactivates the monocyte chemotactic factor was also released from the apoptotic cells 30 hours after the heat treatment. A super infusion of this protease into the skin where the apoptotic cells had been injected diminished the number of infiltrated monocytes. The present results indicate an important role of the S19 ribosomal protein dimer in the phagocytic clearance of apoptotic cells.     

Dbp5p, a cytosolic RNA helicase, is required for poly(A)+ RNA export

The DBP5 gene encodes a putative RNA helicase of unknown function in the yeast Saccharomyces cerevisiae. It is shown here that Dbp5p is an ATP-dependent RNA helicase required for polyadenylated [poly(A)+] RNA export. Surprisingly, Dbp5p is present predominantly, if not exclusively, in the cytoplasm, and is highly enriched around the nuclear envelope. This observation raises the possibility that Dbp5p may play a role in unloading or remodeling messenger RNA particles (mRNPs) upon arrival in the cytoplasm and in coupling mRNP export and translation. The functions of Dbp5p are likely to be conserved, since its potential homologues can be found in a variety of eukaryotic cells.     

Fabrication of a new substrate for atomic force microscopic observation of DNA molecules from an ultrasmooth sapphire plate

A new stable substrate applicable to the observation of DNA molecules by atomic force microscopy (AFM) was fabricated from a ultrasmooth sapphire (alpha-Al2O3 single crystal) plate. The atomically ultrasmooth sapphire as obtained by high-temperature annealing has hydrophobic surfaces and could not be used for the AFM observation of DNA. However, sapphire treated with Na3PO4 aqueous solution exhibited a hydrophilic character while maintaining a smooth surface structure. The surface of the wet-treated sapphire was found by x-ray photoelectron spectroscopy and AFM to be approximately 0.3 nm. The hydrophilic surface character of the ultrasmooth sapphire plate made it easy for DNA molecules to adhere to the plate. Circular molecules of the plasmid DNA could be imaged by AFM on the hydrophilic ultrasmooth sapphire plate.     

Fractal characteristics of gas-solids flow in a circulating fluidized bed

A fractal approach is adopted to describe the dynamic behavior of a circulating fluidized bed. Two times series, differential pressure fluctuations along the riser height and solids momentum fluctuations along the radial direction, are measured and analyzed in terms of fractal dimensions. The influences of operating conditions and axial/radial positions on the fractal dimension are discussed. Attempts are also made to interpret the flow structure in the bed in terms of the fractal dimension. It is found that fractal analysis can provide a useful tool for understanding the characteristics of gas-solids flow in circulating fluidized beds.     

Systematic analysis of glycosphingolipids in the human gastrointestinal tract: Enrichment of sulfatides with hydroxylated longer-chain fatty acids in the gastric and duodenal mucosa

The composition of the glycosphingolipids of the human gastrointestinal tract was studied. The major neutral glycosphingolipids were ceramide monohexosides (e.g., GalCer, GlcCer), LacCer, Gb₃Cer, Gb₄Cer and more polar ones with more than four sugars, whereas neither Gg₃Cer nor Gg₄Cer were present. The acidic glycosphingolipids consisted of sulfatides and gangliosides such as G_M3, G_M1, G_D3 and G_D1a. Also a large amount of sulfatides was found in the gastric mucosa and duodenum. The concentrations of sulfatides in the fundic mucosa, antral mucosa and duodenum amounted to 416.0, 933.8 and 682.9 nmol/g of dry weight, respectively, exceeding those in the gastric mucosa and kidney of other mammals. The major molecular species of the sulfatides were identified as I³SO₃-GalCer with hydroxylated longer-chain fatty acids based on the analyses by gas-liquid chromatography and negative ion fast-atom bombardment mass spectrometry. In contrast, gangliosides in these regions showed a tendency to be lower than sulfatides, and the molar ratios of sulfatides to gangliosides were about 2.0, whereas those in other parts were less than 0.5. A high content of sulfatides in the gastric and duodenal mucosa, where mucosa is easily insulted by acid, pepsin and bile salts, may be closely related to their roles in mucosal protection. The nomenclature used for gangliosides and other glycosphingolipids follows the system of Svennerholm (Ref. 1) and the recommendation of the IUPAC-IUB Commission (Ref. 2), respectively.     

Comparative study of lipogenic enzymes in several vertebrates

The liver lipogenic enzymes are compared among rats, chickens, frogs and fish. Although the apparent Km values of glucose-6-phosphate dehydrogenase for glucose-6-phosphate are not much different among all the species, those of malic enzyme for malate are much higher in chickens and fish than in rats and frogs. Glucose-6-phosphate dehydrogenase showed very high activities compared with malic enzyme in fish liver, and malic enzyme showed high activities in chicken liver. Although the apparent Km values of acetyl-CoA carboxylase and fatty acid synthetase for substrates are in the same range among all the animals, the activity of acetyl-CoA carboxylase seems to be extremely low in fish and frog livers, and that of fatty acid synthetase is low in frog livers only. In addition, the apparent Km values of α-glycerophosphate acyltransferase of fish liver are very high, and the enzyme activity appears to be extremely low compared to the others. Therefore, the enzymes at the first steps of both fatty acid and glycerolipid syntheses of poikilothermos animals appear to be very low. On the other hand, the Ouchterlony double-diffusion patterns showed that the lipogenic enzymes of chickens, frogs and fish are immunologically different from those of rats, with the exception of acetyl-CoA carboxylase in chickens. Therefore, it is suggested that the fatty acid and glycerolipid forming systems of poikilothermos animals are quite different from those of homoiothermos and the lipogenesis is very low in poikilothermos.     

High oleic acid oil suppresses lung tumorigenesis in mice through the modulation of extracellular signal-regulated kinase cascade

This study was undertaken to estimate the effect of dietary high oleic acid oil (OA) on 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced lung tumorigenesis in mice. Diet containing 10% oil was fed to mice through experimental periods. On day 30 after NNK injection (100 mg/kg body weight, i.p.), the treatment increased the level of prostaglandin E₂ (PGE₂) as well as proliferating cell nuclear antigen, a marker of cell proliferation in a high linoleic acid oil (LA)-fed group but not in an OA-fed group. The NNK treatment also induced the activation of an extracellular signal-regulated kinase (Erk) cascade (Erk, Mek and Raf-1) in an LA-fed group. On the other hand, OA feeding abolished the NNK-induced activation of the Erk cascade. In conjugation with these events, OA feeding reduced lung tumor incidence and tumor multiplicity (percentage of mice with tumors) in mice compared with LA feeding at the 20th experimental week. These results suggest that OA suppresses lung tumorigenesis and that this suppression is correlated with the inhibition of PGE₂ production and inactivation of the Erk cascade.