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Biomedical subjects

I Horii

Publications and source records attributed to I Horii.

At least 55 records · Page 3Linked to original sources
PubMed1992

Stratum corneum sphingolipids and free amino acids in experimentally-induced scaly skin.

Stratum corneum sphingolipids are of particular importance in maintaining the water permeability barrier of mammalian epidermis. Free amino acids also play an important role in water retention in the stratum corneum. To clarify the way in which these substances affect scaly skin, stratum corneum sphingolipids and free amino acids collected from artificially-induced scaly skin were analysed. Scaly skin was induced by tape stripping. The total amount of sphingolipids was quantified by gas chromatography and five of sphingolipid fractions were isolated and quantified by thin-layer chromatography. Free amino acids were analysed using a high-speed amino analyser. The total amount of sphingolipid in scaly skin did not differ statistically from that in control skin. However, a significant change in the distribution of the five sphingolipid species was observed in scaly skin and the total amount of amino acids was decreased in scaly skin. These results suggest that the distribution of these five types of sphingolipid and the total amount of amino acids are responsible for scaly skin.

Adult↗
PubMed1992

Different stability of ligand-receptor complex formed with two endothelin receptor species, ETA and ETB.

There are at least two types of endothelin receptors, ETA and ETB, present in various tissues. We found that although biotinylated ET-1 could bind to both ETA and ETB receptors, the stability of the formed ligand-receptor complexes was different. When the preformed complexes of receptor (solubilized from canine brain and lung membranes) and biotinylated ET-1 were subjected to avidin agarose column chromatography, most of the ETA activity was recovered in the pass-through fraction and the remainder was recovered in the 0.5 M KSCN eluate as ligand-free forms. On the other hand, the ETB activity bound firmly to the avidin agarose column was eluted with 1.5 M KSCN. The detection of the complex of 125I-ET-1 and its receptor by SDS-PAGE run at a low temperature was only possible with the ETB fractions and the complex of 125I-ET-1 and ETA was unstable during the separation. These results suggest that the conformation of the ligand binding sites of canine ETA and ETB as well as the stability of their ligand-receptor complexes to SDS are significantly different. Similar observations were also obtained for human ETA and ETB receptors.

Animals↗
PubMed1992

Effect of 1,25-dihydroxyvitamin D3 on the female reproductive system in rats.

Parenteral application of the active metabolite of vitamin D3, 1,25-dihydroxyvitamin D3, has been anticipated to have remarkable efficacy in secondary hyperparathyroidism. However, in a reproduction seg. I study in rats, poor reproductive performance was reflected in a decrease in the number of matings, implantations and live births. These changes were though reversible after treatment with the compound was discontinued. In order to clarify the mechanism of these reversible toxicities, the following were examined in female rats treated with the D3 metabolite: 1. effect on the estrous cycle (no treatment for 2 weeks, treatment for 3 weeks and recovery for 2 weeks), and 2. effect on the maintenance of pregnancy (treatment for 2 weeks before mating and during the gestation period). In both groups, the levels of calcium, calcitonin, PTH and progesterone in serum were measured, and histopathological examination of the thyroid, parathyroid, ovary and uterus was carried out. The following results were observed: 1) disturbance of the estrous cycle, 2) hypofunctional changes in the corpus luteum in the ovary, and the epithelium, endometrium and uterine gland in the uterus with a decrease in the serum progesterone level and 3) hypercalcemia with a decrease in calcitonin or PTH levels in serum with morphological changes including atrophy and cyst-formation in the parathyroid. However, the above changes were reversible, and recovery was observed after administration of the compound was discontinued. These results indicate that the hypercalcemia caused by 1,25-dihydroxyvitamin D3 disrupts endocrinological homeostasis which in turn temporarily disrupts the female reproductive system. Furthermore, it was suggested that 1,25-dihydroxyvitamin D3 itself directly influences on endocrinological organs (hypothalamus, pituitary, parathyroid and thyroid) and reproductive organs (ovary and uterus).

Animals↗
PubMed1992

Effect of hyaluronate on physicochemical and biological properties of collagen solution which could be used as collagen filler.

The effect of hyaluronate (HA) on the physiochemical and biological properties of collagen solution was examined for two preparations of collagen with different rates of fibril formation. The addition of HA to the collagen preparation with the slower rate of fibril formation caused a prominent acceleration of fibril formation. A differential scanning calorimetric measurement of the collagen preparation demonstrated a stabilizing effect of HA on collagen solution after incubation at 37 degrees C. Histochemical examination of rat dermis after injection of the collagen solution into the tissue revealed the migration of fibroblast-like cells into the region occupied by the injected collagen. The addition of HA to collagen preparation S (slower rate of fibril formation) shortened the time-to-appearance of fibroblast-like cells to a similar value to that observed when collagen preparation F (faster rate of fibril formation) was used. The timing of cell appearance was in accord with the rate of fibril formation in vitro. Fibrils newly formed by injected collagen might provide sites for cell attachment, migration and proliferation.

Animals↗
PubMed1991

Detection of an endothelin-1-binding protein complex by low temperature SDS-PAGE.

We found that the complex of ET-1 and its binding protein was stable enough to be separated by SDS-PAGE when electrophoresis was run at a low temperature. Cross-linking was not necessary for the detection of [125I]-ET-1 and its binding protein complex by autoradiography. This simple method could be used in qualitative (estimation of apparent molecular weight of ET-1 binding protein) and quantitative (determination of relative content of ET-binding protein) analysis of the ET-binding protein complex. ET-binding protein complexes of various animal species and organs were investigated by this method.

Animals↗
PubMed1991

In vitro comparison of water-holding capacity of the superficial and deeper layers of the stratum corneum.

We measured the electrical conductance at 3.5 MHz of a sheet of normal stratum corneum (SC) mounted with either the upper surface or the lower surface downward in simulated in vivo conditions. In this way, we assessed the water-binding capacity of the upper and lower portions of the horny layer. Measurements were made with the SC model in environments of various relative humidities. Between 30% and 90% relative humidity, the conductance of the upper surface was always significantly higher than that of the lower surface. In contrast no significant difference was observed in transepidermal water loss (TEWL) with the SC sheet placed upwards or downwards. After exposure to lipid extraction with acetone/ether, a significant decrease in conductance and increase in TEWL occurred, particularly in the upper surface. The amount of extractable amino acids was much higher from the middle layer than from the outer layers of the SC, and was lowest from the deepest part of SC. These results, indicating a lower efficiency of the lower surface of the SC for water-binding than the desquamating upper surface, suggest that newly formed immature SC does not have the water-holding capacity of the mid portion of the SC, which is probably the layer with the greatest water holding capacity.

Adolescent↗
PubMed1991

Response to growth factors of human dermal fibroblasts in a quiescent state owing to cell-matrix contact inhibition.

Mitogenic responses to various growth factors were compared for quiescent human dermal fibroblasts cultured under three different conditions; serum depletion, cell-cell contact inhibition and cell-matrix contact inhibition. The non-dividing fibroblasts cultured under a low serum condition (0.2% fetal bovine serum, FBS) or in a confluent culture with 10% FBS resumed multiplying upon exposure to any one of or any combination of the growth factors examined; epidermal growth factor (EGF), basic fibroblast growth factor (b-FGF), platelet-derived growth factor (PDGF) and transforming growth factor beta (TGF-beta). The only exception was the lack of effect of TGF-beta on the cell under a low serum condition. In contrast, the proliferation of fibroblasts which were growth-arrested in contracted collagen gel by cell-matrix contact inhibition was not stimulated by any of the growth factors examined except for PDGF. It is currently accepted that the mechanism of growth stimulation or signal transduction after binding of each growth factor to the specific receptor depends on the kind of growth factor. The results suggest that the signal transductions delivered by EGF, b-FGF or TGF-beta are inactivated by a high level of interaction of collagen fibrils with the cell membrane (under the condition of cell-matrix contact inhibition); whereas the signal transduction by PDGF is unaffected. The finding supports the existence of a specific growth stimulation pathway for PDGF.

Adult↗
PubMed1991

Heterogeneity of endothelin receptor.

The complex formed between endothelin (ET) and its binding protein was adequately stable to be separated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) at low temperature. Cross-linking was not necessary. This simple method was applied for both qualitative (determination of molecular weight of ET binding protein) and quantitative (determination of content of ET binding protein) analyses of ET-binding protein for various organs from different mammals. Molecular weights of the major ET binding proteins were around 35 and 55 kDa in several canine organs examined. The distribution of these protein species among organs was quite different. In the case of human placenta, the predominant one showed a molecular weight of 35 kDa. We obtained several monoclonal antibodies that could immunoprecipitate ET binding activity from the solubilized human placenta membrane fraction. One of the monoclonal antibodies recognized approximately 58-kDa protein, as determined by Western blot analysis, when the gel is run in the absence of beta-mercaptoethanol.

Animals↗
PubMed1991

Tyrosinemia II: report of an incomplete case and studies on the hyperkeratotic stratum corneum.

We report a case of tyrosinemia II in a 20-year-old man who had painful palmoplantar hyperkeratotic lesions and corneal erosion. His serum tyrosine level was more than 10 times above normal. Though dietary control failed to eliminate the skin lesions, etretinate cleared them temporarily and grafted skin from the buttock did not become hyperkeratotic during the observation period of 1 1/2 years. Amino acid analysis in the hyperkeratotic stratum corneum disclosed a moderate increase not only in soluble tyrosine but also in other amino acid contents. Turnover time of the hyperkeratotic stratum corneum was greatly elongated as compared with that of the grafted site.

Adult↗
PubMed1991

Effect of cysteine on bovine serum albumin (BSA) denaturation induced by solar ultraviolet (UVA, UVB) irradiation.

Long-term exposure to natural sun-light (UVA, UVB) induced fluorescence and caused disulfide bond formation in bovine serum albumin (BSA). The addition of cysteine enhanced the bond formation to such an extent that a solution of BSA was transformed into an insoluble gel. The disulfide bonds in the gels are derived from internal-SH groups of protein. This reaction occurred even if cysteine was added after exposure to ultraviolet (UV)-irradiation. Fluorescent substances seem to be involved in this reaction. On the other hand, low concentrations of cysteine (less than 5 mM) inhibited both fluorescence and disulfide bond formation. The addition of glutathione to BSA produced the same effect as that of cysteine. The addition of thiourea to BSA solution inhibited fluorescence, but did not inhibit disulfide bond formation. We assume that external-SH compounds such as cysteine and glutathione, which have high reactivity with hydroxyl radicals (.OH), act not only as free-radical scavengers, but also as radical mediators in the polymerization of protein through disulfide cross-links induced by UV-irradiation. Solar UVA as well as UVB irradiation are shown to have the same effect on the protein polymerization.

Cysteine↗
PubMed1991

Functional analyses of the superficial stratum corneum in atopic xerosis.

Dermatologists universally recognize that the unaffected skin of patients with atopic dermatitis tends to be dry and slightly scaly. To characterize the functional properties of the superficial stratum corneum in atopic xerosis, we studied the forearms of 28 patients with atopic dermatitis, aged 14 to 30 years, and 18 age-matched controls, with the use of mainly noninvasive methods. Patients with atopic xerosis showed markedly higher transepidermal water loss and markedly lower skin surface hydration levels than did the controls. The corneocytes in atopic xerosis tended to desquamate in clumps of cell aggregates instead of as individual cells. They contained a substantially lower amount of water-soluble amino acids, which play a role in the water-retaining capacity of stratum corneum, than did those of controls. Although the number of stratum corneum cell layers in atopic xerosis (21 +/- 4) was substantially larger than that in controls (15 +/- 1), its turnover time (7 +/- 2 days) was appreciably shorter than that for controls (14 +/- 2 days). Like those noted in the skin with increased epidermal proliferation, the size of superficial corneocytes in patients with atopic xerosis was substantially smaller than in controls. Histopathologic examination revealed acanthotic epidermis, mild perivascular mononuclear cell infiltrate, and pigment incontinence. Atopic xerosis, the dry skin of patients with atopic dermatitis, shows various stratum corneum functional impairments, probably reflecting increased epidermal proliferation due to a low-level ongoing dermatitis.

Adolescent↗
PubMed1990

Experimental cancer cachexia induced by transplantable colon 26 adenocarcinoma in mice.

The present study investigates a tumor model for cachectic mice. Among various murine transplantable tumors, used for assessing cytostatics, we identified colon 26 adenocarcinoma (colon 26) as capable of causing cachexia. Fifteen days after inoculation, the tumor grew to about 6% of the body weight causing substantial carcass weight loss of 3.4 g (14.5% of the carcass weight). When the tumor size was 2.7 g at 3 weeks after the inoculation, the carcass weight was 12 g less than the age-matched control. The tumor continued to grow while the mice maintained this weight, surviving for an average of 45 days. This extensive weight loss was essentially the wasting of adipose and muscle tissues. Hypoglycemia and hypercorticism occurred during the time of the weight loss. In addition, the colon 26 caused disorders of hepatic functions: the concentration of acute phase proteins in serum increased; the number of hepatic glucocorticoid-cytosol receptors decreased; and activities of hepatic catalase and drug-metabolizing enzymes decreased. On the other hand, noncachectic mice with Meth A fibrosarcoma gained weight, which was somewhat less than the control, and had neither hypoglycemia nor hypercorticism, although some mild disorders of hepatic functions were found. Mice bearing colon 26 is an appropriate model for elucidating the mechanism that causes cachexia.

Adenocarcinoma↗
PubMed1990

A distinct characteristic of the quiescent state of human dermal fibroblasts in contracted collagen gel as revealed by no response to epidermal growth factor alone, but a positive growth response to a combination of the growth factor and saikosaponin b1.

It is known that fibroblasts cultured in a reconstituted collagen gel contract the gel, resulting in a high density of collagen fibrils comparable to that in dermis. Our previous study indicated that human fibroblasts in the contracted collagen gel did not proliferate in the presence of 10% serum even though there was no apparent cell-cell contact. We interpreted this cell growth inhibition as being caused by a high level of cell-collagen fibril interactions or cell-matrix contact inhibition. In the present study, the effect of epidermal growth factor (EGF) on fibroblast proliferation in the contracted collagen gel was compared with that on cells in other quiescent states. Non-dividing cells at confluency on a plastic dish or on collagen gel responded to the added EGF and multiplied, while the cells in the contracted collagen gel did not show any growth response to EGF at concentrations up to 100 ng/ml. Binding assay of [125I]-EGF to the cells showed that the number of binding sites and the binding constant obtained from Scatchard analysis were essentially unchanged in the contracted collagen gel, indicating that EGF receptors were not masked by collagen fibrils but that the signal transduction after binding of EGF was blocked. The block in the signal transduction was suppressed by the addition of saikosaponin b1. These results suggested that the quiescent fibroblasts in the contracted collagen gel were in a distinct state from previously known quiescent states of cultured cells, namely quiescent states due to cell-cell contact inhibition at confluency or to deficiency of growth factors. The mechanism of the effect of saikosaponin b1, which has a potent saponin activity, is discussed.

Adult↗
PubMed1989

Demonstration of neutrophil chemotactic anaphylatoxins in human dandruff.

In contrast to scales collected from the scalps of nine healthy individuals where a few parakeratotic cells are observable, a large number of parakeratotic cells associated with some infiltrated polymorphonuclear leukocytes (PMNLs) were found in the scales obtained from 11 individuals complaining of dandruff. Therefore, we determined the neutrophil chemotactic properties of the water-soluble extracts of dandruff scales and normal control scalp scales. Aqeous extracts fractionated by Sephadex G-75 showed a potent chemotactic activity only in the fractions of the dandruff patients that eluted with cytochrome C marker (cyt C; molecular weight, 12 kDa). It was comparatively stable to heat but was greatly inhibited by the addition of anti-C5 antiserum. Radioimmunoassay demonstrated that, although small amounts of C5a and C4a anaphylatoxins were demonstratable even in the extracts of normal scalp, they were found in significantly increased amounts in the extracts of dandruff. Moreover, there was a significantly positive correlation between C5a and C4a concentrations in these extracts. These results suggest that classical complement pathway activation with resultant production of C5a anaphylatoxin is involved in the migration of PMNLs into the lesional skin of dandruff.

Adult↗
PubMed1989

Stratum corneum hydration and amino acid content in xerotic skin.

The relationship between clinical severity, the hydration state of the skin surface as assessed by a conductance with a 3.5 MHz high frequency impedance meter, and the amino acid content of the stratum corneum (SC) of six patients with ichthyosis vulgaris and 30 elderly persons with varying degrees of xerosis was investigated. With an increase in the severity of xerosis the SC showed a decrease in hydration and in its extractable amino acid content. There was a significant correlation between the hydration state and amino acid content of the SC. Although there was a significant correlation between the amino acid content of the lower leg SC and that of the forearm SC in the same subject, the former was generally lower, corresponding to the greater incidence of xerosis on the lower leg. These results suggest that a decreased amount of amino acids may be the result of low profilaggrin biosynthesis in the epidermis and that this is involved in the pathogenesis of these xerotic skin conditions. Clinical improvement of the xerosis following treatment with a urea-containing cream was not accompanied by any significant change in the amino acid content of the SC.

Aged↗
PubMed1989

[Synthesis of 2-oxooctahydroimidazo[1,2-alpha]pyridine-3-spiro-4'-piperidine derivatives possessing antiapomorphine activity].

4'-Carbamoyl-1,4'-bipiperidine 1 was dehydrogenated on Pd-C to give 2-oxo-2,3,5,6,7,8-hexahydroimidazo[1,2-alpha]pyridine-3-spiro-4'-p iperidine 2, which was reduced to 2-oxo-1,2,3,5,6,7,8,8a-octahydroimidazo[1,2-alpha]pyridine-3-spiro -4'- piperidine 3 with NaBH4. The iminodibenzyl and similar derivatives of 2 and 3 were synthesized and evaluated by using antiapomorphine test in mice. The derivatives of 3 had more potent antagonistic activity than those of 2 and the order of potency of 3 was: chloriminodibenzyl greater than chlordibenzocycloheptene greater than iminodibenzyl greater than iminostilbene greater than fluorene. Further chemical modification of the most active chloriminodibenzyl derivative 8, such as the substitution of Cl atom to Br atom, N-methylation on amine moiety, the replacement of imidazopyridine ring to imidazopyrrole ring, did not give any positive effect. Therefore, 8 may have potential usefulness as an antipsychotic drug.

Animals↗
PubMed1983

Histidine-rich protein as a possible origin of free amino acids of stratum corneum.

The origin of free amino acids and/or their metabolites of the stratum corneum was investigated by pulse-chase experiments using hairless mice. Two, 24, 48, and 72 hrs after the animals were pulse-chased with 3H-histidine or 3H-arginine, radioactivity was determined in the following three fractions of the epidermis: the 0.1 N HClO4 soluble, ethanol soluble fraction (Fr. I); the 0.1N NClO4 soluble, ethanol insoluble fraction (Fr. II); and the 0.1 N HClO4 insoluble, 8M urea soluble fraction (Fr. III). Radioactivity of the epidermal proteins was also determined in SDS-PAGE gels by the gel slicing method. At first, 3H-histidine and/or 3H-arginine were predominantly incorporated into Fr. III, especially into a certain epidermal protein which showed little mobility on SDS-PAGE. Subsequently, 3H, once incorporated into Fr. III, appeared to shift from Fr. III to Fr. II and, simultaneously, from the top band(s) to a band of 32,000 daltons. As a result of amino acid analysis, a protein of Fr. II was considered a histidine-rich protein corresponding to HRP-II of the new born rats of Ball et al. After 48 hrs, as the radioactivities of Fr. III and Fr. II decreased remarkably, Fr. I gradually increased in radioactivity. The major radioactive substances in Fr. I at 72 hrs were identified as 3H-histidine and 3H-urocanic acid when 3H-histidine was used as a tracer, and as 3H-arginine, 3H-ornithine, and 3H-citrulline in the case of 3H-arginine. The amino acid composition of the histidine-rich protein was very similar to that of the free amino acids of the stratum corneum in the hairless mouse, in which some amino acid metabolites were considered as their precursor amino acids. These results strongly suggest that the free amino acids and/or their metabolites of the stratum corneum might be the final products of a degradation of the histidine-rich protein.

Amino Acids↗