Determination of glucosylceramide contents in crop tissues and by-products from their processing.

Glucosylceramides were surveyed in crop tissues and by-products from their processing. Apple pulp contained the highest amount (0.94 mg g(-1)) of glucosylceramide and relatively less sterylglucoside, which is the major contaminant of partially purified glucosylceramide. Glucosylceramide from apple pulp was principally composed of 2-hydroxypalmitic acid as the fatty acid, 4-hydroxy-cis-8-sphingenine as the sphingoid base, and glucose as the hexose, similar to those of commercial preparations isolated from rice bran or wheat germ. Apple pulp may be an alternative source for the commercial production of glucosylceramides.

Role of the pectinolytic enzyme in the lactic acid fermentation of potato pulp by Rhizopus oryzae.

Rhizopus oryzae strain NBRC 4707 produced lactic acid and ethanol more efficiently than strain NRRL 395 in potato pulp, an agricultural by-product of the starch industry. The two strains developed comparable activities of xylanase, cellulase, alpha-amylase, and glucoamylase, while the polygalacturonase activity of strain NBRC 4707 was double that of strain NRRL 395. The addition of commercial pectinase enhanced the formation of metabolites, suggesting that the degradation of pectic substances determines the fermentation of potato pulp by R. oryzae. Orange and apple peel were more effective in the induction of polygalacturonase activity than potato pulp, sugarbeet pulp, or wheat bran when used as a principal carbon source for fungal growth in a solid-state culture. The fungal cells in both types of fruit peel stimulated the fermentation of potato pulp and increased the quantity of lactic acid and ethanol to higher levels than those in other agricultural by-products.

Glomangiopericytoma causing oncogenic osteomalacia. A case report with immunohistochemical analysis.

A 47-year-old woman suffered from gait disturbance due to back pain and muscle weakness. Laboratory data showed serum hypophosphatemia, elevated alkaline phosphatase, and a normal level of ionized calcium. Radiological examinations revealed multiple pathologic fractures in the ribs and pubic rami. She had had no episode of familial or any other notable disorder, and so she was initially treated with medication for adult-onset osteomalacia. However, 19 years later (when she was 66 years old), she noticed a soft-tissue tumor in her buttock. The tumor was excised. The histological features were those of glomangiopericytoma characterized by both glomus tumor-like and hemangiopericytoma-like structures. After removal of the tumor, her symptoms disappeared immediately. Laboratory data normalized 8 months later. To our knowledge, this is the first report of oncogenic osteomalacia caused by glomangiopericytoma.

Presacral ganglioneuroma arising in an elderly man with persistent constipation.

Presacral ganglioneuroma in a 70-year-old man with persistent constipation and its features on computed tomography, magnetic resonance, and endoscopic ultrasonography are presented. Barium enema study and laparotomy showed that constipation was caused mainly by extrinsic compression from this tumor. Computed tomography, magnetic resonance, endoscopic ultrasonographic features such as well-defined solid tumor with a cystic component and punctate calcifications may facilitate early diagnosis of this rare tumor.

(10E,12Z,15Z)-9-hydroxy-10,12,15-octadecatrienoic acid methyl ester as an anti-inflammatory compound from Ehretia dicksonii.

The methanol extract of Ehretia dicksonii provided (10E, 12Z, 15Z)-9-hydroxy-10,12,15-octadecatrienoic acid methyl ester (1) which was isolated as an anti-inflammatory compound. Compound 1 suppressed 12-Otetradecanoyl-phorbol-13-acetate (TPA)-induced inflammation on mouse ears at a dose of 500 microg (the inhibitory effect (IE) was 43%). Linolenic acid methyl ester did not inhibit this inflammation at the same dose. However, the related compounds of 1, (9Z,11E)-13hydroxy-9,11-octadecadienoic acid (5) and (9Z,llE)13-oxo-9,11-octadecadienoic acid (6), showed potent activity (IE500 microg of 63% and 79%, respectively). Compounds 1, 4 ((9Z, 12Z, 14E)-16-hydroxy-9,12,14-octadecatrienoic acid), 5 and 6 also showed inhibitory activity toward soybean lipoxygenase at a concentration of 10 microg/ml.

Spontaneous vertebral arteriovenous fistula--case report.

A 57-year-old male presented with a rare case of spontaneous vertebral arteriovenous fistula manifesting as radiculopathy of the right arm, subsequently associated with pulsating tinnitus and vascular bruit in the nape. He had a past history of chiropractic-induced vertebrobasilar infarction. Angiography showed a simple and direct fistula between the third segment of the right vertebral artery and the epidural veins at the C-1 level, where the artery runs backward above the arch of the C-1 just proximal to the penetration of the dura. The fistula was successfully obliterated by coil embolization, resulting in rapid improvement of the signs and symptoms. Mechanical compression to the nerve roots by the engorged epidural veins with arterial pressure was considered to be the major cause of radiculopathy. Vertebral artery dissection induced by chiropractic manipulation is most likely responsible for the development of the fistula.

Influence of finishing on the electrochemical properties of dental alloys.

Dental alloy surface finishing procedures of may influence their electrochemical behavior, which is used to evaluate their corrosion resistance. We examined the polarization resistance and potentiodynamic polarization profile of the precious-metal alloys, Type 4 gold alloy and silver-palladium alloy, and the base-metal alloys, nickel-chromium alloy, cobalt-chromium alloy, and CP-titanium. Three types of finishing procedure were examined: mirror-finishing using 0.05 micron alumina particles, polishing using #600 abrasive paper and sandblasting. Dissolution of the alloy elements in 0.9% NaCl solution was also measured and compared with the electrochemical evaluation. The corrosion resistance of the dental alloys was found to relate to finishing as follows: The polarization resistance and potentiodynamic polarization behavior revealed that the corrosion resistance improved in the order of sandblasting, #600-abrasive-paper polishing, and mirror-finishing. While the corrosion potential, critical current density and passive current density varied depending on the type of finishing, the transpassive potential remained unchanged. The influence of finishing on the corrosion resistance of precious-metal alloys was less significant than on that of base-metal alloys. A mirror-finishing specimen was recommended for use in evaluation of the corrosion resistance of various dental alloys.

Expression and signal transduction of calcium-sensing receptors in cartilage and bone.

We previously showed that Ca2+-sensing receptors (CaRs) are expressed in chondrogenic RCJ3.1C5.18 (C5.18) cells and that changes in extracellular [Ca2+]([Ca2+]o) modulate nodule formation and chondrogenic gene expression. In the present study, we detected expression of CaRs in mouse, rat, and bovine cartilage and bone by in situ hybridization, immunocytochemistry, immunoblotting, and RT-PCR; and we tested the effects of CaR agonists on signal transduction in chondrogenic and osteogenic cell lines. In situ hybridization detected CaR transcripts in most articular chondrocytes and in the hypertrophic chondrocytes of the epiphyseal growth plate. Expression of CaR transcripts was weak or absent, however, in proliferating and maturing chondrocytes in the growth plate. In bone, CaR transcripts were present in osteoblasts, osteocytes, and bone marrow cells, but rarely in osteoclasts. A complementary DNA was amplified from mouse growth plate cartilage, which was highly homologous to the human parathyroid CaR sequence. Immunocytochemistry of cartilage and bone with CaR antisera confirmed these findings. Western blotting revealed specific bands (approximately 140-190 kDa) in membrane fractions isolated from growth plate cartilage, primary cultures of rat chondrocytes, and several osteogenic cell lines (SaOS-2, UMR-106, ROS 17/2.8, and MC3T3-E1). InsP responses to high [Ca2+]o were evident in C5.18 cells and all osteogenic cell lines tested except for SaOS-2 cells. In the latter, high [Ca2+]o reduced PTH-induced cAMP formation. Raising [Ca2+]o also increased intracellular free [Ca2+] in SaOS-2 and C5.18 cells. These studies confirm expression of CaRs in cartilage and bone and support the concept that changes in [Ca2+]o may couple to signaling pathways important in skeletal metabolism.

Preparation and properties of novel gramicidin S analogs possessing a tri-, tetra- or pentamethylene bridge between ornithine side chains.

Gramicidin S (GS) analogs in which the Ndelta atoms of the two Orn side chains are linked by an oligomethylene bridge [-(CH2)n-; n=3-5] were prepared via the bis(p-nitrobenzenesulfonyl) derivative [Orn(NBS)2,2']GS. For comparison the nonbridged secondary amino group-containing analog [Orn(Me)2,2']GS was also prepared. 1H NMR and CD spectral analysis indicated that these analogs adopt the same beta-sheet conformation as GS. The antimicrobial activities of these analogs were very similar, but were slightly dependent on the bridge chain length, the trimethylene-bridged analog being the most potent.

Structures of the deoxy and CO forms of haemoglobin from Dasyatis akajei, a cartilaginous fish.

The three-dimensional structures of the deoxy- and carbonmonoxyhaemoglobin (Hb) from Dasyatis akajei, a stingray, have been determined at 1.6 and 1.9 A resolution, respectively. This is one of the most distantly related vertebrate Hbs to human HbA. Both structures resemble the respective forms of HbA, indicating that the alpha2beta2-type tetramer and the mode of the quaternary structure change are common to Hbs of jawed vertebrates. Larger deviations between D. akajei Hb and human HbA are observed in various parts of the molecule, even in the E and F helices. Significant mutations and/or conformational changes are also observed around the haems, in the C-terminal region of the beta subunit, in the alpha1beta2 interface and in the organic phosphate-binding site of HbA. Despite these structural differences, the oxygen affinity, haem-haem interaction, Bohr effect and organic phosphate effect of D. akajei Hb are all only moderately reduced. Compared with human HbA, the overall r.m.s. deviation of main-chain atoms in the helical regions of bony fish Hbs is smaller than that of D. akajei Hb.

Analysis of carbohydrate residues on recombinant human thyrotropin receptor.

An investigation of the sugar groups on recombinant human TSH receptors (TSHR) expressed in CHO-K1 cells and solubilized with detergents is described. Western blotting studies with TSHR monoclonal antibodies showed that the receptor was present principally as two bands with approximate molecular masses of 120 and 100 kDa. Further blotting studies using lectins and/or involving treatment with different glycosidases indicated that the 100-kDa band contained about 16 kDa of high mannose-type sugars, and the 120-kDa band contained about 33 kDa of complex-type sugars. It was possible to separate the 120- and 100-kDa components of the TSHRs by lectin affinity chromatography. In particular, Galanthus nivalis lectin, which binds high mannose-type sugars, bound the 100-kDa band, but not the 120-kDa band, whereas Datura stramonium lectin, which binds complex-type sugars, bound the 120-kDa band, but not the 100-kDa band. 125I-Labeled TSH binding studies with the various lectin column fractions showed that TSH-binding activity was principally associated with the complex-type sugar containing the 120-kDa form of the receptor rather than the high mannose-containing 100-kDa form. During peptide chain glycosylation, high mannose-type sugar residues are attached first and then modified by the formation of complex type structures to form the mature glycoprotein. Our data suggest that in the case of the TSH receptor, this type of posttranslational processing has an important role in forming the TSH-binding site.

Clinical diversity in acute necrotizing encephalopathy.

Acute necrotizing encephalopathy is a novel disease entity, proposed by Mizuguchi et al in 1995, that shows a characteristic selective and symmetric involvement of the thalamus, brain stem, and cerebellum. It usually leaves sequelae. The etiology of acute necrotizing encephalopathy is unknown. We describe here six patients aged 6 months to 5 years (four boys and two girls). Four cases were typical, and the patients' cranial computed tomographic scans and magnetic resonance imaging showed irreversible symmetric involvement of the thalamus, brain stem, and cerebellum. Three of the patients died, and one was left with severe sequelae. In the other two patients, who had selective reversible thalamic involvement, the disease was mild; one also showed transient unilateral thalamic involvement. These patients recovered completely. We consider the illness in these two patients to fit the criteria of the mild form of acute necrotizing encephalopathy. We believe that acute necrotizing encephalopathy has some clinical diversity, as is seen in other neurologic disorders, and that a mild form could exist.

Effects of cyclic loading on the strength of all-ceramic materials.

PURPOSE: To investigate the effects of fatigue on the strength of materials used in all-ceramic crowns, the biaxial flexural strength of all-ceramic restorative materials was measured with precracked and laminated specimens after cyclic loading. MATERIALS AND METHODS: Two types of all-ceramic systems were used to prepare specimens: a glass-infiltrated alumina core system (In-Ceram) and a leucite-reinforced feldspathic porcelain system (IPS-Empress). Monolayer and laminated disks with a diameter of about 11.75 mm and a thickness of 1.20 +/- 0.05 mm were prepared. The biaxial flexural strength of the specimens that were polished and/or created with a precrack was measured. Their strength was also measured following cyclic loading. A cyclic load that was 60% of the mean breaking load of the specimens (before cyclic loading) was applied to specimens for 10(5) cycles in 37 degrees C water. RESULTS: Although 20% to 30% of the polished specimen samples fractured during cyclic loading, the biaxial flexural strength of specimens that survived the cyclic loading was nearly the same as that of specimens not subjected to the cyclic loading. The strength of the alumina system decreased with the introduction of precracks, and nearly all specimens fractured during cyclic loading. The strength of the leucite system, however, did not decrease with the presence of precracks, and no fractures were observed on these specimens during cyclic loading. CONCLUSION: These results suggest that although the alumina system has high flexural strength, it is more sensitive to flaws and susceptible to fatigue fracture. The effect of fatigue on the leucite system appears to be low.

Hemiballismus associated with Influenza A infection.

A case of 2-year and 3-month-old boy developing right hemiballismus and transient selective unilateral involvement in the left thalamus on MRI during Influenza A infection is reported. Hemiballismus is an extremely rare neurological complication and, to our knowledge, this is the first case showing hemiballismus during Influenza A infection.

Simultaneous determination of nicotinic acid and its metabolites in rat urine by micellar electrokinetic chromatography with photodiode array detection.

Nicotinic acid, nicotinamide and their possible metabolites were successfully separated within 17 min by micellar electrokinetic chromatography using 50 mM borate buffer (pH 9.0) containing 150 mM sodium dodecyl sulfate as the running buffer. Calibration curves for all compounds showed good linearity in a range of 5 microg/ml and 250 microg/ml with good correlation. The present method did not require any clean-up procedures and made it possible to determine all metabolites without interference on a photodiode array detector. Urine samples collected from Wistar male rats were analyzed after high-dose oral or intravenous administration of nicotinic acid or nicotinamide. Metabolic pathways of nicotinic acid in male Wistar rats are also discussed.

Human choline acetyltransferase mRNAs with different 5'-region produce a 69-kDa major translation product.

Choline acetyltransferase (ChAT, EC 2.3.1.6) is the biosynthetic enzyme for acetylcholine. We have previously shown that multiple ChAT mRNA species with different 5'-noncoding regions are expressed in the rat and mouse. However, the diversity of ChAT mRNA species in human has not completely been elucidated. In this work N1- and N2-type ChAT cDNAs were cloned from a human brain cDNA library and the N-exon located in the human ChAT gene. Polymerase chain reaction analysis indicates that four species of ChAT mRNAs (R-, N1-, N2- and M-types) are produced in human brain and spinal cord. In all human transcripts, the ATG initiation codon in the rat, mouse and pig was replaced by ACG, which does not serve as an initiation codon for translation. In vitro translation and mammalian expression analyses revealed that N1-, N2- and R-type mRNAs give rise to a single 69 kDa enzyme, while M-type mRNA produces both 82 and 69 kDa enzymes. The translation efficiency of M-type mRNA was lower than that of the other mRNA species. Moreover, the translation efficiency of human ChAT mRNAs was considerably lower than that of rat ChAT mRNA, suggesting that the ATG codons for human ChAT are unfavorable for translation initiation compared with the initiation codon for rat ChAT. These results provide rational explanations for the previous reports that human ChAT protein purified from the brain and placenta had 66-70 kDa molecular mass, and that ChAT activity in a single motor neuron of human was far lower than that of other vertebrates. Sequencing of monkey ChAT gene showed that the initiation ATG in rodent ChAT was also replaced by ACA in the monkey.

Surface reaction between titanium castings and investments.

The selection of investments suitable for titanium castings is critical because the surface-reacted layer greatly affects their mechanical properties. This study analyzed the interfacial zone of titanium castings obtained from various commercial investments and examined the relationship between the surface-reacted layer and the hardness. CP titanium disks (20 mm diameter, 1 mm thick, n = 5) were cast using eight commercial casting systems and investments for titanium casting: two alumina-based investments (T-invest, CD-titan investment), two spinel-based investments (Titanmold, Titavest MZ), three zirconia-based investments (Asahivest-D, Titaniumvest EX, Tycast investment TM), and a magnesia-based investment (Selevest CB). The Vickers hardness of cast disks in cross-section was tested at 50 microns intervals from the surface to 0.5 mm in depth. The cross-section was observed metallurgically, and an elemental mapping measurement of the reacted layer was conducted under the EPMA. The Vickers hardnesses at 50 microns from the surface of the specimens ranged 312 to 624 and were significantly different among the investments (p < 0.01). The thickness of the reacted layer, rich in O, Si, Al, Zr, Mg, and P, ranged from 10 to 60 microns. No correlation was seen between the thickness of reacted layer and the hardness (p > 0.05). However, the specimens cast with alumina-based and zirconia-based investments were harder than the spinel-based and magnesia-based investments at 500 microns. Although the free energies of metal oxide formation of alumina and zirconia were lower than titania, these data suggest that silica, alumina and zirconia were reduced by contact with molten titanium. Accordingly, silicon, aluminum and zirconium diffused to the interior of the titanium castings and changed their mechanical properties.

Apoptosis induced by selenium in human glioma cell lines.

Several studies have shown that selenium can inhibit tumorigenesis in tissues. However, little is known about the mechanism and the effect of selenium on DNA, especially in brain tumor cells. In this study we examined the biological effect of selenium on human glioma cell lines (A172 and T98G). Selenium exhibited an antiproliferative effect on these cell lines (and induced the typical ladder pattern of DNA fragmentation commonly found in apoptosis), which were prevented by catalase. Few effects of selenium on NT14 fibroblasts were found. These findings demonstrate that selenium may induce, by apoptosis, cell death of human glioma cell lines, which are resulting from free radical oxygen forming.

Immunohistochemical study on collagenous proteins and biophysical analysis of crystals in extraskeletal chondroma.

The immunohistological distribution of collagen types I, II, III, and VI in five cases of extraskeletal chondroma was examined and compared with that in six cases of enchondroma. In addition, the composition of crystals deposited in three cases of extraskeletal chondroma were biophysically analyzed with special attention to the relationship between the collagen types of the matrix and the crystal deposition. In extraskeletal chondroma, immunoreactivity of type II collagen in the extracellular matrix and type VI collagen in the pericellular area, which were strongly and diffusely recognized in the normal hyaline cartilage and enchondroma, was diminished. Instead, additional types of collagen, types I and III, were demonstrated in the matrix. Electron roentgenographic microanalysis and infrared light spectroscopic analysis revealed that calcium pyrophosphate dihydrate (CPPD) was included in the crystals of extraskeletal chondroma. CPPD crystals were observed in/around collagen types I and III. The possible relationship between the difference of collagen composition in the matrix and the CPPD crystal deposition is discussed.

Effect of selenium on malignant tumor cells of brain.

Some reports have demonstrated that selenium can inhibit tumorigenesis in some tissues of animal. However, little is known about the inhibitory effect on malignant tumor cells of brain. The purpose of our study was to determine the biological effect of selenium on growth of rat glioma and human glioblastoma cell lines. Cell lines C6 and A172 were obtained from Japanese Cancer Research Resources Bank, Tokyo, Japan (JCRB). Cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal calf serum at 37 degrees C in a humidified atmosphere of air and 5% CO2. Antiproliferative effects of selenium were evaluated using growth rate assay quantifying cell number by MTT assay. An antiproliferative effect of selenium was found in two cell lines, which was more effective on human A172 glioblastoma and less effective on rat C6 glioma.