Complete regression of low-grade mucosa-associated lymphoid tissue (MALT) lymphoma in the gastric stump after eradication of Helicobacter pylori.

Recent studies have suggested that Helicobacter pylori (H. pylori)-associated gastritis may play an important role in the pathogenesis of primary gastric lymphoma. Recently, triple therapy using proton pump inhibitor, amoxicillin, and clarithromycin, has been established for the eradication therapy of H. pylori infection, and is also recommended for the treatment of the superficial type of low-grade gastric MALT (mucosa-associated lymphoid tissue ) lymphoma. MALT lymphoma of the gastric stump is rare, and total resection or chemotherapy for MALT lymphoma of the gastric stump has been previously reported. Therefore, there is no evidence that eradication therapy is effective for low-grade MALT lymphoma of the gastric stump. Our case illustrates the remarkable efficacy of eradication of H. pylori for low-grade MALT lymphoma of the gastric stump without other modalities such as surgery and systemic chemotherapy.

Depigmenting effect of alpha-tocopheryl ferulate on normal human melanocytes.

Oral vitamin E supplementation has been reported to improve facial hyperpigmentation. alpha-Tocopheryl ferulate (alpha-TF) is a compound of alpha-tocopherol (alpha-T) and ferulic acid connected by an ester bond. Ferulic acid is also an antioxidant, and could scavenge free radicals induced by ultraviolet (UV) radiation, and thus maintain the long-lasting antioxidative effect of alpha-T. Previously we have reported that alpha-TF inhibited melanogenesis in human melanoma cells. To know whether alpha-TF might be useful as a whitening agent to improve and prevent facial hyperpigmentation, the depigmenting effect of alpha-TF in normal human melanocytes was examined in this study. The results showed that 30 microg/ml of alpha-TF dissolved in 150 microg/ml of lecithin inhibited melanization significantly without inhibiting cell growth. This phenotypic change was associated with the inhibition of tyrosinase and the degree of inhibition was dose dependent. No significant effect on DOPAchrome tautomerase (DT) activity was observed. These results suggest that alpha-TF is a candidate for an efficient whitening agent which suppresses melanogenesis. In this paper, the role of alpha-T and alpha-TF in inhibiting biological reactions induced by reactive oxygen species (ROS) is also discussed.

The depigmenting effect of alpha-tocopheryl ferulate on human melanoma cells.

Oral vitamin E (alpha-tocopherol, alpha-T) supplementation has been reported to improve facial hyperpigmentation. alpha-Tocopheryl ferulate (alpha-TF) is a compound of alpha-T and ferulic acid connected by an ester bond; ferulic acid is also an antioxidant, and could scavenge free radicals induced by ultraviolet (UV) radiation, and thus maintain the long-lasting antioxidative effect of alpha-T. Our aim was to see whether alpha-TF might be useful as a whitening agent and an antioxidant to improve and prevent facial hyperpigmentation following UV exposure. In this study, the inhibitory effect of alpha-TF on melanogenesis was examined biochemically using human melanoma cells in culture. The results show that alpha-TF, solubilized in ethanol or in 0.5% lecithin, inhibited melanization significantly, as did alpha-T at a concentration of 100 microg/mL, without inhibiting cell growth. This phenotypic change was associated with inhibition of tyrosinase and 5, 6-dihydroxyindole-2-carboxylic acid polymerase activities, and the degree of inhibition was dose dependent. No significant effect on DOPAchrome tautomerase activity was observed. alpha-TF did not directly inhibit tyrosinase activity of the large granule fraction extracted from human melanoma cells, and Western blotting revealed that there were no changes in protein content or in molecular size of tyrosinase, tyrosinase-related protein (TRP)-1 or TRP-2. Therefore, the inhibition of tyrosinase activity by alpha-TF might be due to effects at the post-translational level, and possibly by a secondary molecule activated by alpha-TF. These results suggest that alpha-TF is a candidate for an efficient whitening agent which suppresses melanogenesis and inhibits biological reactions induced by reactive oxygen species.

Modulation of melanocyte-stimulating hormone receptor expression on normal human melanocytes: evidence for a regulatory role of ultraviolet B, interleukin-1alpha, interleukin-1beta, endothelin-1 and tumour necrosis factor-alpha.

Melanocyte-stimulating hormone (MSH) receptor binding activity and melanocortin-1 receptor (MC1-R) gene expression on normal human melanocytes have been studied as responses to the effects of ultraviolet B (UVB), interleukin-1 (IL-1), endothelin-1 (ET-1) and tumour necrosis factor-alpha (TNF-alpha), which are known as UV sensitive regulators of melanocytic function. MSH receptor (MSH-R) binding activity was upregulated by UVB, IL-1alpha, -1beta and ET-1, but was downregulated by TNF-alpha. Northern blot analysis showed that MC1-R mRNA expression was induced 24 h after UVB irradiation in a dose-dependent manner, and that 24-h treatment with ET-1 also induced an expression of MC1-R mRNA, whereas TNF-alpha downregulated the expression. In addition, IL-1alpha and -1beta have a small but real inductive effect on MC1-R mRNA expression. Taken together, our results suggest a model in which higher MC1-R mRNA expression is accompanied by upregulation of MSH-R binding activity, and enhanced by UVB or cytokines sensitive to UVB. Such a regulatory system would enable normal human melanocytes to respond to MSH more efficiently and induce an increase of melanization of the skin through the MSH/MSH-R system after UVB radiation.

Effect of arbutin on melanogenic proteins in human melanocytes.

The inhibitory effect of arbutin, a naturally occurring beta-D-glucopyranoside derivative of hydroquinone, on melanogenesis was studied biochemically by using human melanocytes in culture. Cells were cultured in the presence of different concentrations of arbutin. The maximum concentration of arbutin that was not inhibitory to growth of the cells was 100 micrograms/ml. At that concentration, melanin synthesis was inhibited significantly by approximately 20% after 5 days, compared with untreated cells. This phenotypic change was associated with the inhibition of tyrosinase and DHICA polymerase activities, and the degree of inhibition was dose dependent. No significant difference in DOPAchrome tautomerase (DT) activity was observed before or after arbutin treatment. Western blotting experiments revealed there were no changes in protein content or in molecular size of tyrosinase, TRP-1 or TRP-2, indicating that inhibition of tyrosinase activity by arbutin might be due to effects at the post-translational level.

Developmental changes in carbohydrate antigens in embryonic rat lens.

The functions of glycosphingolipids, especially those containing the alpha-galactosyl epitope, were investigated during the development and differentiation of rat lens. Glycosphingolipids in embryonic lens tissue were mainly composed of neolacto-series glycosphingolipids and sialic acid-containing ganglio-series gangliosides GM3 and GD3. These glycosphingolipids and gangliosides were widely expressed on cell membranes in the lens vesicle and the elongating lens fibers. In particular, the expression of neolacto-series glycosphingolipids with the alpha-galactosyl epitope was found to be associated with the differentiation and interaction of lens fibers. Glycoproteins with the alpha-galactosyl epitope was also involved in the elongation of lens fibers. The expression of the glycoproteins was highly specific in elongating lens fibers when these were examined in head sections obtained at various embryonic stages. Thus, the alpha-galactosyl epitope on glycosphingolipids and glycoproteins appears to be associated with the differentiation and elongation of lens fibers in the rat. Evolution-related changes in the expression of carbohydrate antigens are also discussed in relation to the development and cell-to-cell interaction of lens fibers in mammals.

Identification and synthetic pathway of sialyl-Lewis(x)-containing neolacto-series gangliosides in lens tissues. 2. Enzymatic synthesis of sialyl-Lewis(x) gangliosides in monkey and rat lenses.

In Japanese monkey lenses, 3H-labeled fucose and N-acetylneuraminic acid were enzymatically transferred to neolactotetraosylceramide (nLc4) and III 3 FucnLc4, respectively, suggesting the presence of a synthetic pathway of IV3 NeuAcIII3 FucnLc4 via III3 FucnLc4 in monkey lenses. Six rat strains, Wistar, Sprague-Dawley and pigmented strains, contained sialyl-Lewis(x) gangliosides in non-cataractous lenses in a strain-specific manner. Glycosyltransferase assay revealed that the transfer of 3H-labeled fucose to nLc4 occurred in all the strains, but that the transfer of 3H-labeled N-acetylneuraminic acid to III3 FucnLc4 was strain-specific. These results suggested that sialyl-Lewis(x) gangliosides were generally synthesized from neolactotetraosylceramide via Lewis(x) glycolipid (III3 FucnLc4) in lens tissues, differing from other tissues. Combining our results, we propose two synthetic pathways of sialyl-Le(x)- containing neolacto-series gangliosides and A-pathway ganglio-series gangliosides in human senile cataractous lens: one to sialyl-Lewis(x) gangliosides from nLc4 via Lewis(x) glycolipid, and the other to GD1a from GM3, via GM2 and GM1.

Identification and synthetic pathway of sialyl-Lewisx-containing neolacto-series gangliosides in lens tissues. 1. Characterization of gangliosides in human senile cataractous lens.

Human lens accumulates gangliosides in association with aging and senile cataract progression. In this study we purified and characterized five major gangliosides in human cataractous lenses. Structural analyses and immunological studies revealed the presence of ganglio-series gangliosides, GM3, GM2, GM1 and GD1a, and a sialyl-Lewisx-containing neolacto-series ganglioside, NeuAc alpha 2-3Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1ceramide (IV3NeuAcIII3FucnLc4). Slow-moving gangliosides, although minor components, were also found to have sialyl-Lewisx-related structures, based on anti-Lewisx antiserum binding to their asialo forms. However, sialyl-paragloboside, a possible precursor of the sialyl-Lewisx ganglioside, was not identified.

Age-related changes in ganglioside composition in human lens.

We previously reported that human lens accumulates gangliosides in association with aging and senile cataract progression. Structural analysis revealed that gangliosides in human cataractous lenses were composed of ganglio-series gangliosides, such as GM3, GM2, GM1 and GD1a, and sialyl-Lewisx-containing neolacto-series gangliosides. Although Lewisx-containing neolacto-series glycolipid was found to accumulate in association with aging and cataract progression, the sialyl-Lewisx gangliosides did not show much accumulation in individual lenses from subjects between 16 and 80 years old. The content of sialyl-Lewisx gangliosides was about two to four times higher than that of Lewisx glycolipids, suggesting the possibility that the increase in Le(x) glycolipid is partly due to the desialylation of sialyl-Le(x) gangliosides. On the other hand, the expression of ganglio-series gangliosides increased in an age-related manner. Thus, age-related changes in lens glycolipids may modify the cell-to-cell interaction induced by cell surface sugar chains, leading to the initiation and progression of cataract.

Localization of neutral and acidic glycosphingolipids in rat lens.

Rat lens was found to contain several neutral and acidic glycosphingolipids in lens epithelia, cortex and nucleus, and showed developmental changes in their content and localization. TLC-immunostaining of gangliosides revealed the enrichment of some ganglio-series gangliosides (GM3, GM1, GD3 and GD1b) in lens epithelia and the presence of GM3 and GD3 in the lens nucleus. Immunohistochemical studies confirmed the distribution of GM3 and GM1 in anterior lens epithelial cells and the cortex, with expression decreasing toward the lens nucleus. Immunoreaction to GD3 was more intense in the lens nucleus than in epithelial cells. In contrast, the expression of neolacto-series glycosphingolipids was restricted to the lens nucleus. In order to investigate the pathological changes of glycosphingolipids in cataract, galactose-induced cataractous lenses were examined. However, no significant changes were observed in the content and composition of glycosphingolipids. In addition, Lewisx epitopes found in human cataractous lenses were not detected in the cataractous lenses of galactosaemic rats and hereditary cataractous Emory mice.

Characterization of neutral glycosphingolipids in rat lens.

Neutral glycosphingolipids were purified from non-cataractous lenses of Sprague-Dawley rats by a combination of solvent extraction, Folch's partition, and column chromatography using DEAE-Sephadex and Iatrobeads. Six major GSLs from monohexosylceramide to pentahexosylceramide were identified by sugar composition analysis, methylation analysis and glycosidase digestion. Structural relationships among the six neutral glycosphingolipids revealed metabolic pathways leading to the synthesis of Gal alpha 1-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1 ceramide (IV3Gal alpha nLc4), instead of a Lewis(x) glycolipid (Gal beta 1- 4(Fuc alpha 1-3)GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1 ceramide, III3FucnLc4), from neolactotetraosylceramide (nLc4), together with isoglobotriaosylceramide (iGb3). The alpha-galactosyl epitope, Gal alpha 1-3Gal-R, is evolutionarily conserved in many types of cells of non-primate mammals, prosimians and New World monkeys, but not in those of Old World monkeys or humans. This evolution-related difference in carbohydrate epitopes suggests different cell-to-cell attachments, which may be mediated through cell surface glycosphingolipids, between rat and human lenses.

Alteration in the fluorescence polarization of rat plasma and liver cell membranes following bile duct ligation in rats.

The fluorescence polarization levels of liver cell membranes and plasma were analyzed to determine membrane fluidity following bile duct ligation (BDL) in rats. Fluorescence polarization was measured with a spectrofluorophotometer equipped with polarizers, using 1,6-diphenyl-1,3,5-hexatrien (DPH) as a probe. After bile duct ligation, liver cell membrane fluidity decreased significantly for up to 14 days after surgery (P < 0.001 on 3rd and 7th days). The polarization of the plasma in rats with BDL slightly but significantly increased compared to the levels in the control animals over the 14-day period following BDL. In addition, a small but significant correlation in the polarization levels between plasma and liver cell membranes (r = 0.362, P < 0.02) was observed. The co-incubation of BDL plasma with normal liver cell membranes resulted in a decrease in membrane fluidity, which suggested that BDL rat plasma had a direct effect on membrane fluidity. After a 70% hepatectomy, the polarization of the membranes from remnant livers in the BDL rats remained elevated relative to the sham-operated controls. It is thus concluded that the membrane fluidity of the livers in BDL rats decreases following bile duct ligation and does not increase after a 70% hepatectomy, presumably due to the increased plasma level of bilirubin.

Comparative study of glycosphingolipid composition in mammalian lenses.

The carbohydrate epitope Gal alpha 1-3Gal-R (alpha-galactosyl epitope), which is detectable by its binding with Bandeiraea simplicifolia-IB4 lectin, was found in glycosphingolipids (GSLs), both neutral and acidic (gangliosides), from lens tissues of non-primate mammals, but not in those of human senile cataracts and Old World monkeys. Instead, human cataractous and Old World monkey non-cataractous lenses expressed Lewisx (Le(x)) epitopes (Gal beta 1-4(Fuc alpha 1-3)GlcNAc-R) in neutral GSLs. Sialylated Le(x) epitopes were found in rat and pig lenses as well as in human and Old World monkey lenses. Ganglio-series gangliosides, consisting mainly of GM3, GM1, GD1a and GD3, were detected in a species-specific fashion. On the other hand, alpha-galactosyl epitopes were expressed in lens tissues only in water-insoluble proteins of non-primate mammals, but Le(x) and sialylated Le(x) epitopes were not detectable in lens proteins. Among the several mammalian lenses examined, humans and Old World monkeys showed similar GSL compositions, in particular the presence of Le(x) and sialylated Le(x) epitopes and the absence of alpha-galactosyl epitopes, in lens tissue.

Glycosphingolipids in cultured lens epithelial cells from dog and rhesus monkey.

Vertebrate lens tissues contain several species of acidic and neutral glycosphingolipids in relatively high amounts. However, the epithelia with capsule from dog and rhesus monkey lenses had a simpler composition and lower content of glycosphingolipids than whole lenses. Gangliosides and neutral glycosphingolipids in monolayer cultures of lens epithelial cells were also different from those in whole lenses. Although alpha-galactosyl (Gal alpha 1-3Gal-R) or Lewis(x) (Gal beta 1-4[Fuc alpha 1-3]GlcNAc-R) epitopes were found in glycosphingolipids from whole lenses, they were not detected in those from monolayer cultures of dog and rhesus monkey lens cells. In addition, significant changes in ganglio-series gangliosides were induced in monolayer cultures of both cells, where GM3 and GD3 were predominant. Immunofluorescence study revealed a characteristic distribution of cell surface gangliosides in confluent monolayers. These findings suggest that glycosphingolipid synthesis in lens epithelia is intrinsically different from that in cortical and nuclear fibres, and that the expression of Lewis(x) and alpha-galactosyl epitopes in glycosphingolipids appears to be associated with the differentiation of epithelial cells to fibres.

Characterization of neutral glycosphingolipids in human cataractous lens.

Neutral glycosphingolipids were purified from human senile cataractous lenses by a combination of solvent extraction, Folch's partition, acetylation, and column chromatography using DEAE-Sephadex and Iatrobeads. Six major glycosphingolipids (A-F) from monohexosylceramide to pentahexosylceramide were identified by sugar composition analysis, methylation analysis, secondary ion-mass spectrometry, glycosidase digestion, and chromium trioxide oxidation. Their structures suggested that they were closely related in their metabolism: their sugar chains were in sequence and their ceramide moieties were similarly composed, namely C16:0 and C24:1 constituted most of the fatty acids, and long-chain base components were mostly C18-dihydrosphingosine with a small portion of C18-sphingosine. The sugar chains implied two pathways branching from lactosylceramide: one to globotriaosylceramide and the other to lactotriaosylceramide, which leads to the production of Le(x) glycolipid via neolacto type 2 core chain.

[A case of lamellar keratoplasty and autoscleroplasty combined with keratoepithelioplasty for Wegener's sclerocorneal ulcer].

A 62-year-old man with Wegener's granulomatosis developed sclerocorneal ulcer. The general condition of the patient became well with steroid and presented cyclophosphamide therapy, he developed sclerocorneal ulcer. The sclerocorneal ulcer was present at the nasal limbus of the right eye. The ulcer was marked and presented descemetocele. Although lamellar keratoplasty was performed, the scleral ulcer recurred after two weeks. Excision of the affected sclera and autoscleroplasty was performed. The ulcer disappeared after 6 months, and the corneal graft was covered with corneal epithelium and conjunctiva. Ten months after autoscleroplasty was performed, a sclerocorneal ulcer was again recurred. We performed lamellar excision of the affected corneal area, resected the adjacent conjunctiva and performed keratoepithelioplasty. These surgical procedures were effective in preventing perforation of the cornea. Specular microscopic study of the corneal epithelium revealed nucleated epithelium and large epithelial cells. The endothelium appeared normal.

Senile cataract-related accumulation of Lewis(x) glycolipid in human lens.

A glycosphingolipid that reacted positively to anti-stage-specific embryonic antigen-1 (SSEA-1) antiserum accumulated in human lens in association with aging and senile cataract formation. Since this antiserum recognizes Lewis(x) (Le(x)) structure, Gal beta 1-4(Fuc alpha 1-3)GlcNAc-, which is a typical tumor-associated and differentiation-related saccharide chain, the lens glycolipid was predicted to be a Lex antigen. The glycolipid purified from cataractous lens tissues was indeed a Lex glycolipid, Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3Gal beta 1- 4Glc beta 1-1 ceramide. Enhanced expression of the Lex glycolipid may affect the organization of lens plasma membranes through Le(x)-Le(x) interactions, as suggested for compaction in mouse preimplantation embryos and embryonic teratocarcinomas, resulting in lens opacification, namely cataract.

Increase in lens gangliosides due to aging and cataract progression in human senile cataract.

Gangliosides were isolated from human senile cataractous lenses by solvent extraction, DEAE-Sephadex column chromatography, and thin-layer chromatography. The content and composition of gangliosides were examined in individual lens tissues. Three predominant gangliosides, GM3, GM1, and GD1a, were tentatively identified in comparison with authentic brain gangliosides, and several unidentified gangliosides were also recognized. The increase in ganglioside content per mg of protein content in cataractous lenses was found to be influenced by two physiologic parameters: aging and cataract progression. The mature cataractous lenses showed a higher ganglioside level on a protein basis than the immature lenses compared with the same age group. On the basis of statistical analysis, an age-dependent increase in ganglioside concentration was recognized in both mature and immature lens groups. The relative increase in slow-moving polysialogangliosides on thin-layer chromatography seemed to be caused by the maturation of cataract. The sugar composition of one of the polysialogangliosides was found to be glucose, galactose, and sialic acid in the molar ratio of 2:1:4; this suggests the presence of a unique ganglioside species in human cataractous lens.

Electrical myotonia and cataract in X-linked muscular dystrophy (mdx) mouse.

An X chromosome-linked mouse mutant (mdx) has been investigated as an animal model of Duchenne's muscular dystrophy, and has been found to have the same defect of dystrophin in the muscle surface membrane. Intracellular recordings from the mdx mouse hemidiaphragm preparations revealed low resting membrane potentials and electrical myotonia which occurred at the time of microelectrode insertion and withdrawal. Electrical myotonia of the mdx mouse was observed in 30-50% of the impaled muscle fibers at low temperature, which decreased to only 7.8% at 37 degrees C. Electrical myotonia of mdx mice was not abolished by (+)-tubocurarine. Though there was no behavioral myotonia in mdx mice, repetitive bursts of action potentials in mdx mice were based on the abnormalities of the muscle membrane since neuromuscular blockade did not abolish the repetitive bursts. Also close observation of the lenses of mdx mice revealed cataracts from the newborn stage to the adult age. Slit lamp examination of the lenses of the mdx mice revealed nuclear cataracts followed by anterior subcapsular cataract as they grew. The cataract of mdx mice is different from that of myotonic dystrophy which is usually posterior subcapsular.

Age-dependent changes in monkey lenticular gangliosides.

The content, composition, and distribution of gangliosides were examined in the lenses of normal rhesus monkeys aged 6-16 years. Gangliosides were isolated by organic solvent extraction. DEAE-Sephadex ion-exchange column chromatography, and thin-layer chromatography (TLC). Ganglioside contents determined by the thiobarbituric acid method increased in the lens with aging. TLC analysis of gangliosides showed a much more complex pattern with aging, and the predominant gangliosides were tentatively identified as GM3, GM1, and GD1a. Individual lenticular gangliosides were identified by TLC-immunostaining procedures using anti-GM1 and anti-asialoGM1 antisera.