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Biomedical subjects

H Shinkai

Publications and source records attributed to H Shinkai.

At least 91 records · Page 5Linked to original sources

Human type VI collagen: purification from human subcutaneous fat tissue and an immunohistochemical study of morphea and systemic sclerosis.

Collagen was isolated from human placenta by pepsin digestion and salt precipitation. This collagen was similar in its electrophoretic mobility and immunological reactivity with monoclonal antibody to form B of type VI collagen in the literature (Trueb B, Schreier T, Bruckner P and Winterhalter K. 1987. Eur. J. Biochem. 166: 699-703). We prepared polyclonal rabbit antiserum against alpha 2 chain of type VI collagen and performed an immunohistochemical study using this polyclonal antibody. It reacted in fat tissue and around vessels and peripheral nerves in normal human skin. To confirm the presence of type VI collagen in fat tissue, we isolated collagen from human subcutaneous tissue. This collagen showed a similar pattern in polyacrylamide gel electrophoresis with that from human placenta and cross-reacted with monoclonal or polyclonal antibody against type VI collagen. By immunohistochemical staining, abundant type VI collagen was observed in the septum of subcutaneous fat tissue in morphea or systemic sclerosis. In the mild hyalinizing areas or after treatment with 6M urea or hyaluronidase in highly hyalinized areas, the staining of type VI collagen increased. These data suggest that the amount of type VI collagen in subcutaneous tissue is involved in the early phases of these fibrosing disorders and that type VI collagen accumulates even more in hyalinizing tissue in late phases of these diseases.

Adipose Tissue↗

Pustular psoriasis precipitated by acetazolamide.

We report a case of pustular psoriasis precipitated by acetazolamide, a carbonic anhydrase inhibitor. A 28-year-old man suffering from 20 years-standing generalized pustular psoriasis developed psoriatic arthritis and glaucoma and experienced a precipitation of pustular lesions and widespread erythema after initiation of oral acetazolamide for glaucoma. The cutaneous lesions and systemic symptoms improved after restriction of acetazolamide and administration of oral etretinate. Readministration of acetazolamide confirmed that generalized pustules arose within 24 hours.

Acetazolamide↗

A case of pretibial myxedema associated with Graves' disease: an immunohistochemical study of serum-derived hyaluronan-associated protein.

The myxomatous materials in cutis from a patient with pretibial myxedema (PTM) with Graves' disease were found to be mainly hyaluronic acid (HA), which had accumulated extensively in the upper dermis. Fibroblasts were increased in number in the mid to lower dermis. To clarify the mechanism of the deposition of HA in the dermis, we employed an antibody against serum-derived hyaluronan-associated protein (SHAP). This immunohistochemical study disclosed that the greater part of the fibroblasts in the mid to lower dermis stained positively, while the fibroblasts surrounded by fairly large amounts of HA in the upper dermis stained faintly or not at all. From these results, we suspect that the accumulation of HA progresses from the upper to the low dermis and that the interaction of fibroblasts and SHAP leads to development of the physiophathological cutaneous changes seen in our case.

Fibroblasts↗

Unusual cutaneous manifestations of myelodysplastic syndrome.

We report a 58-year-old patient with acute myeloid leukaemia who developed an oedematous erythema resembling Sweet's syndrome, accompanied by atypical erythema nodosum and bullous pyoderma gangrenosum. Examination of skin biopsies showed dense infiltration with mature neutrophils, although there was peripheral blood leucocytopenia. The oedematous erythema worsened after he was treated with granulocyte-colony-stimulating factor (G-CSF), which was given for his leucocytopenia. We suggest that when a neutrophilic dermatosis complicates leukaemia, alternatives to G-CSF should be considered for the treatment of leucocytopenia. Corticosteroids were very effective in controlling the skin lesions in our patient.

Acute Disease↗

Malignant rhabdoid tumor of the breast: a case report.

A 66-year-old woman developed a malignant rhabdoid tumor of the breast, with a tumor doubling time of 10 days. One month after radical mastectomy, there was local recurrence, followed by multiple pulmonary metastases, and the patient died of respiratory failure 5 months after surgery. The gray-white-colored tumor measured 13 x 12 x 10 cm, and its border was well defined. The tumor was composed of diffusely growing round or polygonal cells with vesicular nuclei, prominent nucleoli, and ample cytoplasm containing eosinophilic inclusions. Lymph node involvement was widespread. Both vimentin and keratin were clearly demonstrated by immunohistochemical staining. Ultrastructural studies revealed that the MRT cells contained cytoplasmic whorls of intermediate filaments.

Aged↗

Connective tissue naevus with pseudo-Hurler polydystrophy.

We report a 39-year-old patient with a huge connective tissue naevus on his back, in association with pseudo-Hurler polydystrophy (mucolipidosis III). As far as we are aware, this is the first report of the coexistence of these two rare conditions. The lesional skin was composed of densely packed, coarse collagen fibres, which were immunohistochemically found to consist of type I, type III and type VI collagens. The amount of elastic fibres was moderately reduced. The glycosaminoglycan content of lesional skin was similar to that in a normal control. Activities of several glycosidases were markedly decreased in cultured fibroblasts.

Adult↗

A new bullous pemphigoid antigen.

Previously we reported a case of subepidermal blistering disease, which resembled bullous pemphigoid clinically. An immunoblot analysis revealed that the patient's serum reacted exclusively with a 450-kD epidermal polypeptide. The 450-kD antigen was also expressed in human keratinocytes and a human squamous carcinoma (HO 1-u-1 or A431) cell line. Using the patient's serum, cDNA clones in a bacteriophage lambda gt11 library derived from human foreskin keratinocytes were immune-screened. One positive clone was isolated from 2 million clones. Northern blot analysis showed that the inserted part of this clone hybridized with a more than 12-kb mRNA species derived from HO 1-u-1 cells. The deduced amino acid sequence had no homology with any other human component.

Autoantigens↗

Three-dimensional fine structure of the lingual papillae and their connective tissue cores in the human tongue.

The three-dimensional structure of the four types of lingual papillae and their connective tissue cores (CTCs) in the human tongue was studied by scanning electron microscopy after removal of the epithelial cell layer, and was compared to those of other mammalian species. Filiform papillae are densely distributed on the dorsal surface of the anterior two-thirds of the tongue. Each filiform papilla has numerous slender protrusions on the top. After removal of the epithelium, the CTC of the filiform papilla has a columnar primary core with 10-30 rod-shaped small secondary cores surrounding an upper central depression. A few long spine-like protrusions are frequently found in the center of this upper depression. The number, thickness and length of each secondary protrusion of the filiform CTC show some variations at the region distributed on the tongue. Fungiform papillae with spherical heads are scattered among these filiform papillae and are numerous at the anterior margin of the tongue. The CTC of the fungiform papilla shows a coralliform structure with numerous small rod-shaped protrusions on the lateral surface, and its branched top has flat areas with a few small round depressions harboring taste buds. It must be emphasized that human fungiform papilla has some taste buds not only in the young, but also in the adult age. Several to 12 vallate papillae are distributed in front of the terminal groove and had the CTC of pinecone-like structure with numerous small thorns (secondary connective tissue cores). Foliate papillae consist of 10-15 parallel folds at the posterior margin of the tongue. The CTC of the foliate papillae appears as ridges and grooves. Small protrusions are scattered on the surface of the ridges. The underside view of the exfoliated epithelium of the vallate as well as the foliate papillae revealed numerous taste buds arranged in single file in the lateral epithelium. The dorsal surface of the root of the tongue has evenly distributed short rod-shaped protrusions of the connective tissue.

Adolescent↗

Structure and localization of O- and N-linked oligosaccharide chains on basement membrane protein nidogen.

The carbohydrate content of mouse nidogen predicts the occupation of two N- and about seven O-linked acceptor sites. The corresponding oligosaccharides were examined by sequential exoglycosidase digestions. The data indicate N-linked substitutions by several bi-, tri- and tetraantennary complex types of oligosaccharides which are further modified by additional lactosamines and terminal alpha-galactose and/or sialic acid. Mannose-rich oligosaccharides were of low abundance. O-linked structures included a di- and tetrasaccharide core structure that were in addition sialylated and may be similar to structures found in fetuin. Evidence is provided that the two sequence-predicted asparagine acceptors are almost fully substituted. Sequence analysis of tryptic peptides identified Thr-271, Ser-303, Thr-309, Thr-317, Thr-320, Thr-892 and Thr-905 as the most likely sites for galactosamine substitutions. These residues are located in the flexible link connecting the N-terminal globular domains G1 and G2 of nidogen and at the border between the rod and the C-terminal globe G3. Four of them showed Pro in the -1 or +3 position. All these Ser, Thr and Pro residues but not the N-linked attachment sites are identical in human nidogen.

Amino Acid Sequence↗

[Pharmacokinetics of MX2, a new morpholino anthracycline, in CSF following intravenous injection].

MX2 x HCl is a new morpholino anthracycline derivative with molecular weight 622.07, and highly lipophilic. In the animal experiments, MX2 was found to cross the blood brain barrier after i.v. injection. Its distribution into the brain was increased by intracarotid injection. In the present study, we examined the distribution of MX2 into the cerebrospinal fluid (CSF) after i.v. administration (5 mg/kg) in normal rabbits. Five min after injection, plasma concentration of MX2 reached to the maximum level (4344.5 ng/ml). CSF concentration of MX2 was at the highest level (75.8 ng/ml) 10 min after injection, and thereafter decreased gradually in parallel with plasma concentration. At 5 hrs after injection, CSF concentration became 26.7 ng/ml, AUC half time of elimination, and mean residence time were 3093.8 ng.hr/ml, 4.57 hrs and 5.10 hrs in plasma and 212.3 ng.h/ml, 5.23 hrs and 7.14 hrs in CSF, respectively. These results indicate that MX2 is able to distribute into CSF after i.v. injection, and expected to be a new anticancer drug for brain and leptomeningeal tumors.

Animals↗

[Pharmacokinetics and antitumor activity of MX2, a new morpholino anthracycline in brain tumor intracerebral transplanted in rats].

Pharmacokinetics and antitumor activity of MX2.HCl (MX2), a new morpholino anthracycline, were investigated in rats transplanted 9L gliosarcoma cells in the brain. (1) Pharmacokinetics: AUC of MX2 in the brain tumors which received intracarotid and intravenous injection of 2mg/kg of MX2 were 117.50 and 55.94 micrograms.hr/g, respectively. AUC of the brain tissue was 1.38-3.90 micrograms.hr/g. (2) Antitumor activity: The inhibition of cell growth at the concentration of 0.1 micrograms/ml was 73.1% with MTT assay. The mean survival time in tumor-bearing rats after intracarotid and intravenous injection of 2mg/kg of MX2 prolonged significantly. Therefore, it seems that MX2 will become an efficacious drug for the treatment of malignant glioma.

Animals↗

A case of subepidermal blister disease associated with autoantibody against 450 kD protein.

We report a case of subepidermal blister disease with eruptions distributed on the trunk and extremities. The blisters regressed after administration of corticosteroid and cyclophosphamide. An immunoblot analysis revealed that the patient's serum reacted exclusively with a 450-kD epidermal polypeptide. It did not react with a 230-kD or a 180-kD bullous pemphigoid antigen. The immunofluorescence study showed the patient's serum reacted with the basement membrane zone of human skin.

Aged↗

Determination of the new morpholino anthracycline MX2.HCl and its metabolites in biological samples by high-performance liquid chromatography.

Methods for determining concentrations of a new morpholino anthracycline MX2.HCl and its metabolites in biological samples using reversed-phase high-performance liquid chromatography and fluorescence detection are described. The limits of detection were less than 1 ng/ml for all compounds after extraction from 0.5 ml of plasma using C18 Sep-Pak cartridges and consecutive solvent extraction. The recoveries from rat plasma ranged from 72.0 to 89.3%. The peak-height ratio of the fluorescence intensities of these compounds versus internal standard showed a linear correlation for concentrations up to at least 500 ng/ml in the plasma (correlation coefficient r greater than 0.999). The within-day and between-day precisions of this assay were in the range 0.8-8.7% (n = 5) and 2.0-3.5% (n = 5), respectively. The concentrations of these compounds in the blood and urine can be also determined by a slight modification of the extraction procedure.

Animals↗

Possibility of ideal blood glucose control by a new oral hypoglycemic agent, N-[(trans-4-isopropylcyclohexyl)-carbonyl]-D-phenylalanine (A-4166), and its stimulatory effect on insulin secretion in animals.

N-[(trans-4-isopropylcyclohexyl)-carbonyl]-D-phenylalanine (A-4166) revealed a new mode of hypoglycemic action with a more rapid onset and a shorter duration of action than the sulfonylureas (SUs). Hypoglycemic mechanisms and glycemic control benefits were demonstrated in laboratory animals. The stimulatory effect of A-4166 on insulin release, in fasting dogs with a cannula into the portal vein, was more rapid than that of tolbutamide after oral administration. A-4166 stopped the stimulation of insulin secretion very quickly, whereas tolbutamide maintained an elevation in plasma insulin levels for at least 6 hours. In the case of A-4166, a counter-regulatory glucagon response was observed during recovery from hypoglycemia, but it was significantly inhibited by tolbutamide. Hyperglycemia induced by glucose loading was rapidly inhibited by A-4166 in normal rats, in genetically diabetic KK mice and in STZ-induced non-insulin-dependent diabetes mellitus (NIDDM) rats. Also, repeated administration of A-4166 for 2 weeks enhanced insulin secretion in the same manner as a single administration in normal rats. In conclusion, A-4166 is a new type of oral hypoglycemic agent, having a rapid and short-term insulin secretory effect and no suppressive effect on the hypoglycemia-induced glucagon response. Oral therapy with A-4166 would be beneficial in supplementing endogenous insulin secretion and would exert ideal glycemic control in NIDDM patients.

Animals↗

Structure of N-linked oligosaccharide chains in the triple-helical domains of human type VI and mouse type IV collagen.

Asparagine-linked oligosaccharides were liberated from pepsin-treated type VI collagen and the 7S domain of type IV collagen by hydrazinolysis and their structures analysed by exoglycosidase treatment. The major component in both proteins was complex biantennary oligosaccharide being partly modified by the addition of fucose and sialic acid residues. The 7S domain contained in addition distinct amounts of truncated biantennary structures lacking one or two beta-galactose residues and a minor triantennary structure. Carbohydrate analysis indicated that all of the N-linked and 80-90% of the O-linked acceptor sites are occupied. The lack of galactosamine content in both collagens showed the O-linked oligosaccharides were only those attached to hydroxylysine and not to serine or threonine. The high carbohydrate density along both triple helical domains is discussed with regard to their limited ability to form lateral aggregates.

Animals↗

Heterogeneity of clinical features of pseudoxanthoma elasticum: analysis of thirteen cases in Oita Prefecture from a population of 1,240,000.

Thirteen cases among 16,260 patients seen at our department during the nine years from 1981 were diagnosed as pseudoxanthoma elasticum (PXE). The sex and age distributions were similar to those so far reported in the literature. There were two cases of the autosomal dominant type. One patient had skin lesions typical of PXE in association with Ehlers-Danlos syndrome. In two of three patients with cutis laxa-like lesions, eruptions of elastosis perforans serpiginosa were present. In three cases, we failed to detect deposition of calcium in typical skin lesions in either an early or a late stage by histochemical staining, in spite of the presence of slightly degenerated elastic fibers.

Adolescent↗