Novel sulfated oligosaccharides containing 3-O-sulfated glucuronic acid from king crab cartilage chondroitin sulfate K. Unexpected degradation by chondroitinase ABC

We prepared a series of oligosaccharides from king crab cartilage chondroitin sulfate K after exhaustive digestion with testicular hyaluronidase, and determined the structures of four tetrasaccharides and a pentasaccharide by fast atom bombardment mass spectrometry, high performance liquid chromatography analysis of chondroitinase AC-II digests, and 500-MHz 1H NMR spectroscopy. The tetrasaccharides shared the common core structure GlcAbeta1-3GalNAcbeta1-4GlcAbeta1-3GalNAc with various sulfation profiles. One structure was GlcAbeta1-3GalNAc(4S)beta1-4GlcAbeta1-3GalNAc(4S), whereas three of them have the following hitherto unreported structures including a novel glucuronate 3-O-sulfate: GlcA(3S)beta1-3GalNAc(4S)beta1-4GlcAbeta1-3GalNAc(4S), GlcAbeta1-3GalNAc(4S)beta1-4GlcA(3S)beta1-3GalNAc(4S), and GlcA(3S)beta1-3GalNAc(4S)beta1-4GlcA(3S)beta1-3GalNAc(4S), where 3S or 4S represents 3-O- or 4-O-sulfate, respectively. The structure of the pentasaccharide was determined as GlcA(3S)beta1-3GalNAc(4S)beta1-4GlcA(3S)beta1- 3GalNAc(4S)beta1-4GlcA. Chondroitinase ABC digestion of the tetrasaccharides with GlcA(3S) at the internal position destroyed the disaccharide unit containing GlcA(3S) derived from the reducing side and resulted in only the disaccharide unit from the non-reducing side. In contrast, these tetrasaccharides remained totally resistant to chondroitinase AC-II. The results indicated that it is necessary to reevaluate the disaccharide composition of chondroitin sulfate poly- or oligosaccharides purified from various biological sources, since they were usually determined after chondroitinase ABC digestion. It is probable that the structures containing GlcA(3S) would not have been detected.     

Preparation of Silicon Carbide and Aluminum Silicon Carbide from a Montmorillonite–Polyacrylonitrile Intercalation Compound by Carbothermal Reduction

Both silicon carbide and aluminum silicon carbide have simultaneously been obtained directly from naturally occurring aluminosilicate by carbothermal reduction for the first time. A precursor of a montmorillonite–polyacrylonitrile (PAN) intercalation compound was heated at 1700°C in Ar. For comparison, montmorillonite–carbon mixtures were similarly heated. α-SiC, β-SiC, and Al₄Si₂C₅ formed from the montmorillonite–PAN intercalation compound. Mainly α-Al₄SiC₄ was obtained with ternary carbides from the montmorillonite–carbon mixtures in addition to a large amount of β-SiC. Hence, aluminum silicon carbide formation was affected by the mixing condition of the starting materials.     

Sol–Gel‐Derived High‐Performance Stacked Transparent Conductive Oxide Thin Films

Single‐ and multi‐layer transparent conductive oxide (TCO) thin films exhibiting high performance, good packing density and low surface/interface roughness are deposited on silica glass substrates by the sol–gel method. The crystal and microstructural properties of the TCO thin films are evaluated as an alternate to films prepared by ultra‐high vacuum deposition. Tin‐doped indium oxide (ITO) thin films produced using a two‐step drying process showed low surface roughness because of dense packing structure not only horizontal but also vertical directions. As a result, electrical conductivity, carrier concentration, carrier mobility, and optical transmittance of 2.3 × 10³ S/cm, 8 × 10²⁰ cm^?3, 18 cm²/Vs, and over 98% at 500 nm, respectively, were achieved. A multilayer ZnO/ITO stacked structure was also fabricated using the sol–gel process. Our findings suggest that solution‐based methods show promise as an alternative to existing ultra‐high vacuum methods to fabricate TCO thin films.     

Molecular characterization of Xenopus embryo heparan sulfate. Differential structural requirements for the specific binding to basic fibroblast growth factor and follistatin

Enzymatic elimination of heparan sulfate (HS) causes abnormal mesodermal and neural formation in Xenopus embryos, and HS plays an indispensable role in establishing the embryogenesis and tissue morphogenesis during early Xenopus development (Furuya, S., Sera, M., Tohno-oka, R., Sugahara, K., Shiokawa, K., and Hirabayashi, Y. (1995) Dev. Growth Differ. 37, 337-346). In this study, HS was purified from Xenopus embryos to investigate its disaccharide composition and binding ability to basic fibroblast growth factor (bFGF) and follistatin (FS), the latter being provided in two isoforms with core sequences of 315 and 288 amino acids (designated FS-315 and FS-288) originating from alternative mRNA splicing. Disaccharide composition analysis of the purified Xenopus HS showed the preponderance of a disulfated disaccharide unit with uronic acid 2-O-sulfate and glucosamine 2-N-sulfate, which has been implicated in the interactions with bFGF. Specific binding of the HS to bFGF and FS-288, the COOH-terminal truncated form, was observed in the filter binding assay, whereas HS did not bind to FS-315, indicating that the acidic Glu-rich domain of FS-315 precluded the binding. The binding of the HS to bFGF or FS-288 was markedly inhibited by heparin (HP) and various HS preparations, but not by chondroitin sulfate, supporting the binding specificity of HS. The binding specificity was further investigated using FS-288 and bovine intestinal [3H]HS. Competitive inhibition assays of the HS binding to FS-288 using size-defined HP oligosaccharides revealed that the minimum size required for significant inhibition was a dodecasaccharide, which is larger than the pentasaccharide required for bFGF binding. The binding affinity of FS to HS increased in the presence of activin, a growth/differentiation factor, which could be inactivated by direct binding to FS. These results, taken together, indicate that the structural requirement for binding of HS to bFGF and FS is different. HS may undergo dynamic changes in its structure during early Xenopus embryogenesis in response to the temporal and spatial expression of various growth/differentiation factors.     

Investigation of electrical properties of a 120 cm PSC using an inner spark pulser

Characteristics of fast pulse propagation in a large planar spark counter (PSC) are simulated using a pulser located inside the spark gap. Besides the main mode of pulse propagation, three undesirable modes are observed. These latter strongly distort the shape of the pulse. Characteristics of these modes and methods to eliminate their effects are demonstrated. We present an electrical design for a 120 cm spark counter along with some of its electrical properties as revealed by measurements made with the inner spark pulser. In the present counter design the charge of a pulse is shared by several neighboring strips, enabling one to measure the transverse position of a spark to a high degree of accuracy.     

Structural phase transitions of methane+ethane mixed-gas hydrate induced by 1,1-dimethylcyclohexane

Methane+ethane+1,1-dimethylcyclohexane+water system was investigated by using Raman spectroscopy and isothermal phase equilibrium measurements under four-phase (gas+aqueous+large guest species+hydrate phases) equilibrium conditions at 288.15 K. The results suggest that three kinds of hydrate structures emerge at 288.15 K in the methane+ethane+1,1-dimethylcyclohexane+water system. The hydrate structure for this system changed from structure-H to structure-I via structure-II with increase in the mole ratio of ethane to methane.     

Antioxidant Effects of Herbal Tea Leaves from Yacon (Smallanthus sonchifolius) on Multiple Free Radical and Reducing Power Assays,Especially on Different Superoxide Anion Radical Generation Systems

Yacon (Smallanthus sonchifolius), a native Andean plant, has been cultivated as a crop and locally used as a traditional folk medicine for the people suffering from diabetes and digestive/renal disorders. However, the medicinal properties of this plant and its processed foods have not been completely established. This study investigates the potent antioxidative effects of herbal tea leaves from yacon in different free radical models and a ferric reducing model. A hot‐water extract exhibited the highest yield of total polyphenol and scavenging effect on 1,1‐diphenyl‐2‐picryl hydrazyl (DPPH) radical among four extracts prepared with hot water, methanol, ethanol, and ethylacetate. In addition, a higher reducing power of the hot‐water extract was similarly demonstrated among these extracts. Varying concentrations of the hot‐water extract resulted in different scavenging activities in four synthetic free radical models: DPPH radical (EC₅₀ 28.1 μg/mL), 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) cation radical (EC₅₀ 23.7 μg/mL), galvinoxyl radical (EC₅₀ 3.06 μg/mL), and chlorpromazine cation radical (EC₅₀ 475 μg/mL). The yacon tea‐leaf extract further demonstrated superoxide anion (O₂^?) radical scavenging effects in the phenazine methosulfate‐NADH‐nitroblue tetrazolium (EC₅₀ 64.5 μg/mL) and xanthine oxidase assay systems (EC₅₀ 20.7 μg/mL). Subsequently, incubating human neutrophilic cells in the presence of the tea‐leaf extract could suppress the cellular O₂^? radical generation (IC₅₀ 65.7 μg/mL) in a phorbol 12‐myristate 13‐acetate‐activated cell model. These results support yacon tea leaves may be a good source of natural antioxidants for preventing O₂^? radical‐mediated disorders.