Thiosugars. XI. Further studies on thiolevoglycosans.
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Glycolates↗
Biomedical subjects
S Tajima
Publications and source records attributed to S Tajima.
[Studies on the origin of the causative agent of Minamata disease. IX. Transition of the pollution of Minamata Bay and its neighbourhood].
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Animals↗
[Scalp avulsion--its therapeutic experience].
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Adult↗
Determination of 1,5-anhydroglucitol in urine by high performance liquid chromatography and an enzyme sensor.
A simple high performance liquid chromatographic method combined with an enzyme sensor has been developed to measure 1,5-anhydroglucitol in urine. The enzyme sensor consists of a hydrogen peroxide electrode and a chitosan membrane of an immobilized pyranose oxidase. As the system does not resist interfering substances, urine samples are first purified by passing them through a two-layer column packed with (1) strongly basic anion (OH- form, the upper layer) and (2) strongly acidic cation (H+ form, the lower layer) exchange resins. 1,5-Anhydroglucitol is efficiently recovered in the flow-through fraction of the column. In this system, the minimum detectable concentration of 1,5-anhydroglucitol is 0.1 mg/L, and the measurable range extends from 0.1 to 60 mg/L. The coefficient of variation values of the within-day and day-to-day precisions are 3.0-6.5% and and 4.4-6.7% respectively, and there is good agreement between the results measured by our method and those obtained by the gas-liquid chromatographic/mass spectrometric method (r = 0.994). The method we have described here has been successfully used to elucidate a mechanism for the reducing 1,5-anhydroglucitol level in the serum and plasma of patients.
Biosensing Techniques↗
Alginate oligosaccharides modulate cell morphology, cell proliferation and collagen expression in human skin fibroblasts in vitro.
Effects of alginate oligosaccharides on cell proliferation and expression of collagen in cultured skin fibroblasts were studied. The oligosaccharides were found to suppress fibroblast proliferation to half the level in control cultures at a dose of 10 mg/ml during a period of 5 days. The inhibition was accompanied by a change in cell shape. The inhibition of cell proliferation was reversible, since depletion of these oligosaccharides led to a recovery of cell motility. Treatment of confluent cells with 10 mg/ml oligosaccharides for 5 days resulted in a reduction in collagen synthesis to one half of that in control cultures and inhibition of steady state levels of alpha1(I), alpha2(I), alpha1(III) and alpha1(VI) collagen mRNAs. These results suggest that alginate oligosaccharides are potential modulators of dermal fibroblasts and may provide a useful tool for the treatment of disorders related to abnormal collagen metabolism.
Alginates↗
Differential expression of the 45 kDa protein (actin) during the dimorphic transition of Sporothrix schenckii.
We investigated the gene expression of a protein during the dimorphic transition from yeast to mycelial form of Sporothrix schenckii. Yeast cells were converted to mycelial cells in Sabouraud glucose broth at 25 degrees C. After 1, 3 and 5 days of culture, the intermediate form of cells between yeast and mycelium was obtained, and after 7 days these cells were morphologically similar to the mycelial cells. Proteins having the molecular weight of 45 kDa were found to by synthesized preferentially by intermediate form of day 7 and mycelial cells. On the other hand, the 45 kDa proteins were predominantly translated by the RNA isolated from the intermediate of day 7 and mycelial cells using in vitro cell-free translation assay. The 45 kDa proteins synthesized by mycelial cells were found to be identical with the 45 kDa translation products directed by the mRNA isolated from the intermediate and mycelial cells by V8 protease peptide mapping. The 45 kDa protein was considered to be actin by Western blot analysis using an anti-actin monoclonal antibody. These results suggest that the intermediate form of day 7 has the same phenotypes in the morphology and biosynthesis of actin as those of mycelial cells. The expression of the actin gene may be involved in the dimorphic transition of S. schenckii.
Actins↗
Neutral red assay in minimum fungicidal concentrations of antifungal agents.
We assayed the fungicidal effects of antifungal agents using neutral red staining. Fungal elements of Trichophyton mentagrophytes and T. rubrum were treated with various concentrations of antifungal agents in 96-well filtration plates and then stained with neutral red. The amount of neutral red incorporated by the surviving viable cells was determined from the automated spectrophotometric readings at 550 nm. The minimum fungicidal concentrations (MFCs) of antifungal agents determined by this assay correlated well with those determined by conventional assay. This newly developed procedure should provide a rapid, reproducible, quantitative, qualitative and semi-automated susceptibility test for determination of the MFCs of the fungicidal agents.
Antifungal Agents↗
Variation of repetitive DNA and its phylogenetic relation in Hynobiidae (Caudata).
Middle repetitive (MR) DNA comprising about 35% of the genomic DNA of Hynobius retardatus was isolated and used as mixed probes. A 1.3-kb Bam HI fragment, the major repeating unit of the extensively amplified sequences in H. retardatus, was cloned, sequenced, and used as a probe. Dot blot hybridization under the conditions of probe excess revealed that the MR DNA sequences were shared among the three salamander genera belonging to the family Hynobiidae. However, a large difference in the amount of common sequences was found between Hynobius and Salamandrella, and large differences in both amounts and sequence similarity were found between Hynobius and Onychodactylus. Even among species belonging to the genus Hynobius, relative contents of sequences common to the H. retardatus MR DNA were 20% to 50%. Partially related sequences to the 1.3-kb sequence seemed to be present in all the species of Hynobius and Salamandrella, but the extensive amplification was found only in H. retardatus. Based on the relative contents of the common MR DNA and previously reported chromosomal banding characteristics, species in the genus Hynobius are classified into at least three groups. An evolutionary aspect of the emergence of the amplified sequence in H. retardatus is discussed.
Animals↗
Histochemical characterization of low density lipoprotein receptors in internalization-defective familial hypercholesterolemia.
We studied the localization of low density lipoprotein (LDL) bound to the receptors on the cultured fibroblasts from a patient (MN) with homozygous familial hypercholesterolemia and a defect in internalization of LDL and compared the localization with normal fibroblasts and with those from another internalization-defective cell, GM2408A. Monolayers of cells were cultured with lipoprotein-deficient human serum, and the cells were incubated with 125I- or ferritin-labeled LDL. The LDL binding was observed by autoradiography or by an electron microscope. Autoradiographs of bound 125I-LDL confirmed that MN's cells could not internalize LDL inside the cell at 37 degrees C. In these cells, ferritin LDL was found mainly in noncoated regions at 4 degrees C; it was not found in endocytic vesicles after incubation at 37 degrees C. Ferritin cores that bound on the surface of the cells from the normal subject, from GM2408A, from MN, and from MN's parents, were counted and quantitatively analyzed at 4 degrees C. In normal cells, 62% of the ferritin cores were bound in the coated pits; in MN's cells, only 11% of the ferritin LDL was found in the coated pits; in the GM2408A cells, 12% of the ferritin LDL was found in the coated pits; in the cells from MN's parents, 40% of the ferritin LDL was found on the coated pits. The results indicate that the internalization defect in MN's cells is the same as that in the GM2408A cells; neither can localize the LDL and LDL receptor complex in coated pits.
Cells, Cultured↗
Levels of proelafin peptides in the sera of the patients with generalized pustular psoriasis and pustulosis palmoplantaris.
The levels of proelafin peptides in the sera of patients with pustulosis palmoplantaris, a unique type of localized pustular psoriasis, and generalized pustular psoriasis were determined by competitive enzyme-linked immunosorbent assays using antibodies against synthetic proelafin polypeptides corresponding to the elastase inhibitor (elafin) and transglutaminase substrate domains. The sera of patients with pustulosis palmoplantaris (9 cases) exhibited a normal range of proelafin peptide levels. The sera of patients with generalized pustular psoriasis (3 cases) showed high titres of proelafin peptide. There were no large differences in the titres between the 2 antibodies. The antibodies for 2 different domains of proelafin showed a similar immunoreactivity for the non-pustular region of the epidermis in all pustulosis palmoplantaris tested. The results indicate that serum proelafin peptides in pustular psoriasis may depend on the extent of the involved area, and that proelafin peptide level in pustulosis palmoplantaris remains normal despite enhanced local expression in the lesional skin. Since the skin lesions of patients with pustulosis palmoplantaris are limited to the palms and soles, enhanced expression of proelafin in the lesional skin may not lead to elevation of proelafin peptides in the serum.
Adult↗
Coordinate expression of membrane type-matrix metalloproteinases-2 and 3 (MT2-MMP and MT3-MMP) and matrix metalloproteinase-2 (MMP-2) in primary and metastatic melanoma cells.
Expression of MMP-2 in melanoma cells has been demonstrated to be involved in the degradation of extracellular matrix during melanoma growth and to correlate with later melanoma metastasis. MMP-2 is considered to be activated by membrane-associated matrix metalloproteinases (MT-MMPs). To know whether MT-MMPs are involved in the activation of MMP-2 in melanoma cells, immunohistochemical studies were performed in primary and metastatic melanoma by use of the antibodies for MT1-MMP, MT2-MMP and MT3-MMP. Expression of MT1-MMP, MT2-MMP, MT3-MMP and MMP-2 in nevocellular nevus (n = 5), dysplastic nevus (n = 2) and juvenile melanoma (n = 3) was undetectable or detected in only a few cells. Superficial spreading melanoma (SSM) (n = 3) and acral lentiginous melanoma (ALM) (n = 3) showed a moderate expression of MT1 approximately 3-MMP. In nodular melanoma (NM) (n = 2) and metastatic melanoma (n = 3), MT1 approximately 3-MMP was more intensely expressed. Double immunofluorescence demonstrated a consistent colocalization of MT2-MMP/MMP-2 and MT3-MMP/MMP-2 in the NM and metastatic melanoma cells. The colocalization of MT2,3-MMP and MMP-2 in nodular and metastatic melanoma cells suggests that MT-MMPs and MMP-2 co-operate in the invasive and metastatic process of melanoma cells.
Humans↗
Acrokeratoelastoidosis associated with nodular scleroderma.
We report a case of acrokeratoelastoidosis associated with nodular scleroderma, a variant of localized scleroderma. The patient showed no clinical and laboratory signs of systemic scleroderma. Because there is a paper describing the association of acrokeratoelastoidosis with systemic scleroderma, coexistence of acrokeratoelastoidosis and nodular scleroderma in the patient suggests that acrokeratoelastoidosis is related to localized nodular scleroderma as well as systemic scleroderma.
Abdomen↗