Biomedical subjects

S Miyauchi

Publications and source records attributed to S Miyauchi.

At least 91 records · Page 5Linked to original sources

PubMed1995

[Quantitative analyses of glycosaminoglycans in tear fluids in normal human eyes and eyes with corneal epithelial disorders].

The existence and pathophysiological role of glycosaminoglycans in the tear fluid in humans was investigated using quantitative analyses of hyaluronic acid and chondroitin sulfate in the tear fluid. The subjects were 42 eyes of 31 normal controls, 9 eyes of 9 patients with superficial punctate keratitis (SPK), and 13 eyes of 13 patients with epithelial defect. After an instillation of 100 microliters saline solution in the conjunctival sac, as much tear fluid as possible was collected from the lower cul-de-sac. The glycosaminoglycans in the tears were then treated with chondroitinase ABC to make fractions of unsaturated disaccharides. The quantities of disaccharides were determined by high-performance liquid chromatography. Concentrations were expressed as nanomoles of unsaturated disaccharides per protein in the tears. The concentrations of hyaluronic acid and chondroitin sulfate in the normal controls were 0.07 +/- 0.12(n mol/mg protein) and 6.91 +/- 3.63 (n mol/mg protein), respectively. The mean concentration of hyaluronic acid was significantly higher in patients with epithelial erosion than in normal controls, whereas the mean concentration of chondroitin sulfate was significantly lower in patients with epithelial erosion than in normal controls. There was no significant difference in the concentration of glycosaminoglycans between the patients with SPK and normal controls. The results of our study suggest that glycosaminoglycans are synthesized and endogenously secreted into the tear fluids and, especially in the case of hyaluronic acid, may play an important role in corneal epithelial wound healing in patients with epithelial erosion.

Adult↗

PubMed1994

Growth and chemotherapeutic response of cells in a hollow-fiber in vitro solid tumor model.

BACKGROUND: Cancer treatments that appear promising in tissue culture are often less effective in solid tumors, in part because of the proliferative and microenvironmental heterogeneity that develops in these tumors as they grow. Heterogeneous tumor models are thus needed for drug screening. PURPOSE: Our goal was to develop and test for drug evaluation a solid tumor model based on cell growth inside biocompatible hollow fibers. METHODS: Building on the experience of Hollingshead and co-workers with a sparse-cell, hollow-fiber tumor model, we tested six human tumor cell lines for in vitro growth inside 450-microns internal-diameter polyvinylidine fluoride fibers and examined them histologically. Human SW620 colon carcinoma cells grown in hollow fibers were also examined using electron microscopy, and their doxorubicin sensitivity was assessed. A colorimetric assay based on sulforhodamine B was adopted to replace the more cumbersome clonogenic cell survival assay. RESULTS: Five of the human tumor cell lines tested grew to confluence, forming heterogeneous in vitro tumors with subpopulations of viable and necrotic cells. For SW620 hollow-fiber tumors, maximum viable cell populations in excess of 10(8) cells/mL were obtained after 8 days of growth. This viable cell density remained roughly constant for 3-4 days, permitting dose-response experiments over this time interval. Tumor cells in hollow fibers were much more resistant to a 4-hour doxorubicin exposure than were tumor cells in monolayers: LC50 values (i.e., the drug concentrations at which the plating efficiency equals one-half the plating efficiency of untreated cells) of 3.5 microM and 0.16 microM were obtained for hollow-fiber tumors and monolayers, respectively. LC50 values decreased when drug exposure time was increased. Results from the colorimetric assay were in agreement with those from the clonogenic assay. CONCLUSION: The successful growth of tumor cells to confluence in hollow fibers and the feasibility of performing in vitro drug dose-response experiments with a relatively easy colorimetric assay demonstrate the potential of the hollow-fiber solid tumor model as a tool for experimental therapeutic research. IMPLICATION: Hollow-fiber solid tumors may prove useful for experimental drug evaluation.

Antineoplastic Agents↗

PubMed1994

Activation energy for permeation of phosphonium cations through phospholipid bilayer membrane.

The conductance caused by translocation of various phosphonium cations across phospholipid (from soybean) bilayer membrane was measured. Phosphonium cations used were tetraphenylphosphonium (TPP+) and triphenylalkylphosphonium cations formulated as (Phe)3-P(+)-(CH2)nCH3 (n = 0-5). The conductance was dependent on voltage applied externally to the membrane in accordance with a theory developed by previous authors. Using the theory, values of beta K(i) were determined, where beta and k(i) are a linear partition coefficient and a rate constant of transmembrane ion transport, respectively. Since beta k(i) depended on the phosphonium ion concentration, values extrapolated to infinite dilution, (beta k(i))0, were determined. Temperature dependence of (beta k(i))0 allowed us to estimate the activation energy of transport, Ea. For TPP+ thermodynamic values obtained were consistent with values calculated by Flewelling and Hubbell [(1986) Biophys. J. 49, 541-552]. When (Phe)3-P(+)-(CH2)nCH3 (n = 0-5) were used, E(a) depended on the odd or even of n. This "odd and even" pattern was observed in a variety of phenomena such as solubility in water, equivalent ionic conductivity in water, and 31P NMR chemical shift.

Biological Transport↗

PubMed1994

Effect of perfusate pH on the influx of 5-5'-dimethyl-oxazolidine-2,4-dione and dissociation of epidermal growth factor from the cell-surface receptor: the existence of the proton diffusion barrier in the Disse space.

The influx clearance (PSinf.MID) of the weak acid 5,5'-dimethyl-oxazolidine-2,4-dione (DMO) was determined by the multiple indicator dilution method with the isolated perfused rat liver under various perfusate pH conditions, ranging from 6.4 to 7.6. Although the pH partition theory predicted an increase in influx clearance of ten times in proportion to the change in the unionized fraction of DMO, there was no measurable change in this value. The effect of medium pH on the steady-state cell/medium concentration ratio (C/M) ratio of DMO was also investigated using isolated hepatocytes. The C/M ratio increased while medium pH decreased, but this change was less marked than predicted by the pH partition theory. Finally the pH dependency of the dissociation rate constant (koff) of epidermal growth factor from its receptor was also investigated using both isolated rat hepatocytes and the perfused rat liver. When the extracellular pH was changed from 6.4 to 5.6, the koff value of isolated hepatocytes increased 44 times, while that of the perfused rat liver increased only 9 times. Therefore, the effect of changing the extracellular pH on pH-dependent dissociation of epidermal growth factor from its cell-surface receptor was less in the perfused liver than in isolated hepatocytes. These findings, in addition to the well-known existence of the Na(+)-H+ exchanger on the sinusoidal membrane and the possible existence of the unstirred water layer in the Disse space, seem to suggest the existence of the proton diffusion barrier in the rat liver, which remains stronger in the perfused liver than in isolated hepatocytes.

Animals↗

PubMed1994

Hyaluronan, cartilage destruction and hydrarthrosis in traumatic arthritis.

The concentrations of hyaluronan (HA) and chondroitin sulfate (CS) in synovial fluids from patients with traumatic arthritis (TA) with and without hydrarthrosis were measured. The CS in synovial fluids was determined as a marker of cartilage destruction by high performance liquid chromotography. The concentration of HA in synovial fluids was lower in patients with hydrarthrosis than in healthy volunteers and patients with TA without hydrarthrosis, whereas the total amounts of HA and CS and the concentration of CS were higher in patients with hydrarthrosis. To investigate the relation between hydrarthrosis and production of HA in synovial tissues, TA synovial tissue biopsies were stained for HA with biotinylated HA binding region. The intensity of HA staining was higher in specimens from patients with hydrarthrosis than in normal and TA without hydrarthrosis specimens. Thus, there may be a correlation between hyperproduction of HA, cartilage destruction and increase in fluid volume in TA.

Adolescent↗

PubMed1994

Oral administration of BOF-A2 to rats with lung transplanted tumors results in increased 5-fluorouracil levels.

A new tumor model was developed in which solid Yoshida sarcoma tissue injected intravenously developed into tumors in the lungs of about 30% (13/42) of the inoculated rats. Histological examination revealed that alveoli were occupied with tumor cells and the tumors were similar to those obtained by subcutaneous inoculation. Using this model, the concentrations of 5-fluorouracil (5-FU) in tumor tissue 12 h after oral administration of 3-[3-(6-benzoyloxy-3-cyano-2-pyridyloxycarbonyl)-benzoyl]-1-ethoxy methyl- 5-fluorouracil (BOF-A2, 36 mumol/kg) or a combination of tegafur and uracil in a molar ratio 1 to 4 (UFT, 50 mumol/kg) were found to be 129.8 ng/g and 19.2 ng/g, respectively. Thus, compared to treatment with UFT, BOF-A2 resulted in higher levels of 5-FU in lung tumor tissues. Increased concentrations of 5-FU should have a superior anti-tumor effect and so BOF-A2 merits clinical trial in the treatment of patients with lung cancer.

Administration, Oral↗

PubMed1994

Inhibition of catabolic pathway of 5-fluorouracil by 3-cyano-2,6-dihydroxypyridine in human lung cancer tissues.

Our studies of the degradation and the phosphorylation of 5-fluorouracil (5-FU) in normal and tumor lung tissues from 10 cases of lung cancer have shown that the phosphorylation of 5-FU in the tumor tissues was about 2- to 3-fold higher than that in normal tissues, and that the degradation of 5-FU in tumor tissues was nearly 6-fold higher than that in normal tissues. BOF-A2 is an anti-neoplastic agent newly synthesized from 1-ethoxymethyl-5-FU and 3-cyano-2,6-dihydroxypyridine (CNDP). The inhibitory effect of CNDP on the degradation of 5-FU in the tumor tissues was potent (IC50 3.9 x 10(-9) M). Thus, BOF-A2 exerts its anti-neoplastic effect on tumors by potentiating the action of 5-FU through inhibition of 5-FU degradation by the CNDP moiety.

Adenocarcinoma↗

PubMed1994

Proliferative activity of epidermal and dermal T lymphocytes in mycosis fungoides.

BACKGROUND: Mycosis fungoides (MF) is histologically characterized by an accumulation of T lymphocytes (T cells) in both epidermis and dermis. However, the mechanism of the formation of Pautrier's microabscess or the maintenance of the epidermal T cells has not yet been clarified. OBJECTIVE: The rate of proliferation of these T cells was investigated. METHODS: The formalin-fixed, paraffin-embedded sections from 9 cases of patch/plaque stage MF were immunostained with proliferating cell nuclear antigen (PCNA). RESULTS: The PCNA-positive T cell nuclei were found both in the epidermis (including Pautrier's microabscesses) and dermis in all the specimens. The mean PCNA indices of the T cells were 20.2 +/- 10.6% (mean +/- SD) within the epidermis and 16.8 +/- 6.5% in the dermis, which was not statistically significantly different (p > 0.05). The ratio of the PCNA index between the epidermis and dermis was 1.3 +/- 0.6, which was not apparently correlated with the duration of the disease (8.4 +/- 5.1 years). CONCLUSION: These findings suggested that in situ proliferation might contribute to the epidermal T cell population resulting in the formation of Pautrier's microabscesses and an increase in dermal T cell volume during the course of the disease.

Abscess↗

PubMed1994

Localization of hyaluronic acid in human articular cartilage.

To demonstrate localization of hyaluronic acid (HA) in articular cartilage of the human femur, biotinylated HA-binding region, which specifically binds HA molecules, was applied to the tissue. In sections fixed by 2% paraformaldehyde-2% glutaraldehyde, HA staining was detected in lamina splendens and chondrocytes in the middle zone. By pretreatment with trypsin, intense HA staining appeared in the extracellular matrix of the deep zone and weak staining in the superficial and middle zones. Moreover, pre-treatment with chondroitinase ABC (CHase ABC) intensely enhanced the stainability for HA in the superficial and middle zones and weakly in the deeper zone. Combined pre-treatment of trypsin with CHase ABC abolished intra- and extracellular staining for HA in all zones. By microbiochemical study, the concentrations of HA and dermatan sulfate were high in the middle zone, whereas those of chondroitin sulfate and keratan sulfate were high in the deep zone. These results suggest that HA is abundantly synthesized in and secreted from the chondrocytes, particularly in the middle zone, whereas it is largely masked by proteoglycan constituents in the extracellular matrix.

Adult↗

PubMed1994

[Quantitative analysis of glycosaminoglycans in tear fluids during corneal epithelial wound healing in rabbits].

We performed quantitative analyses of glycosaminoglycans in the tear fluids in a rabbit wound healing model. We ablated rabbit corneal epithelium with trephine and spatula, and sampled tear fluids before the epithelial ablation, and at 3, 24, 48, and 72 hours after it. After an instillation of 200 microliters saline solution in the conjunctival sac, as much tear fluid as possible was collected from the lower cul-del-sac. The glycosaminoglycans in the tears were then treated with chondoroitinase ABC to make fractions of disaccharides. The quantities of disaccharides were determined with high-performance liquid chromatography as weight per unit protein in the tears. The concentrations of delta Di-HA in the tear fluids at 3 and 24 hours were significantly higher than those before the treatment and returned to the initial value at 72 hours after making the epithelial wound. Among the disaccharides of chondroitin sulfate, delta Di-0S and delta Di-6S showed a significant increase at 3 hours after the treatment but delta Di-4S did not show any significant variation. The results suggest that the glycosaminoglycans in the rabbit tear fluids may play an important role in the corneal epithelial wound healing process.

Animals↗

PubMed1994

[Effect of BOF-A2 on experimental and spontaneous metastasis model of mouse colon 26].

We investigated the effects of BOF-A2 on the metastasis of colorectal cancer using artificial (experimental) and spontaneous metastasis of Colon 26 tumor cells. BOF-A2 suppressed pulmonary nodules in the artificial metastasis of Colon 26 tumor cells into the lung, and the T/C (%) was 53.1% at 20 mg/kg or 8.3% at 40 mg/kg. BOF-A2 also suppressed pulmonary nodules in the spontaneous metastasis of Colon 26 tumor cells into the lung, and the T/C (%) was 33.3% at 40 mg/kg in postoperative treatment, 33.3% at 10 mg/kg, or 16.7% at 20 mg/kg in preoperative treatment. These effects were accompanied by the prolongation of survival time at each dose. From these results, BOF-A2 showed antimetastatic activity and survival effect, and may be useful for adjuvant chemotherapy to prevent cancer from metastasis to the lung.

Adenocarcinoma↗

PubMed1993

Comparative hepatic transport of desglycylated and cyclic metabolites of rilmazafone in rats: analysis by multiple indicator dilution method.

Rilmazafone (RZ) is an orally active sleep inducer which can be activated to its cyclic form (M1) via the labile desglycylated metabolite (DG). In this scheme, RZ is exclusively metabolized to DG and M1 by aminopeptidases in the small intestine. The concentration of M1 in the systemic plasma after oral administration of RZ has been reported to be higher than that observed after administration of M1, due to the lower hepatic extraction of DG than M1 (Koike et al., Drug Metab. Dispos., 16, 609 (1988)). In the present study, the disposition of DG and M1 in rat liver was investigated, using the multiple indicator dilution method. The hepatic availabilities (F) of DG and M1, assessed from the recovery into the hepatic vein, were 0.16 and 0.07, respectively, which was consistent with the previous in vivo finding that the first-pass elimination of M1 was greater than that of DG. The kinetic analysis based on the distributed model showed that the influx (k'1) and efflux (k'2) rate constants for M1 were larger than those for DG, whereas no significant difference in the sequestration rate constant (k'3) was observed between the two ligands. Based on the concept proposed by Miyauchi et al. (J. Pharmacokinet. Biopharm., 15, 25 (1987)), it was suggested that the determinant factor of the hepatic intrinsic clearance was the influx clearance for both ligands, because the values of k'2 for each ligand were much smaller than the respective k'3 values. It was concluded that the higher plasma concentration of M1 after oral administration of RZ than that observed after administration of M1 is due to the fact that the hepatic uptake of DG is lower than that of M1.

Administration, Oral↗

PubMed1993

Membrane transport of tetraphenylphosphonium and its homologues through the planar phospholipid bilayer: concentration dependence and mutually competitive inhibition in membrane passive transport.

The concentration dependence and mutually competitive inhibition in the membrane transport of the lipophilic cations tetraphenylphosphonium (TPP+) and its homologues were studied by use of a planar phospholipid bilayer. The current-voltage characteristics were analyzed by use of a model developed by Ketterer et al. (J. Membr. Biol. 1971, 5, 225-245). The conductance at 0 mV [G(0)], which represents the transport ability, increased with an increase in concentration, and the G(0) values revealed saturation at higher concentrations. The relationship between G(0) and concentration was well fitted to a Michealis-Menten-type equation with a saturable component (maximum conductance [G(0)max] and concentration yielding one-half of G(0)max[Km]). The G(0)max and Km values were calculated to be 4.1 x 10(-8)-242 x 10(-8) S/cm2 and 13-489 microM, respectively, depending on the lipophilic cations used. The G(0) values normalized by the corresponding G(0)max values were plotted against the concentrations normalized by the corresponding Km values. Normalized curves for TPP+ homologues were all superimposed on a single curve. The mechanism for the saturation of G(0) values may involve the adsorption of TPP+ homologues to a planar phospholipid bilayer, because the Km values calculated in the present study were comparable to those for adsorption to phosphatidylcholine liposomes. We further determined the mutual inhibition of membrane transport by these homologues. Tetraphenylmethylphosphonium at 1 mM, a concentration threefold higher than the Km value for this compound, reduced the G(0) for other TPP+ homologues by 38-55%, whereas reduction by 10 mM tetraethylammonium was minimal. The type of inhibition was classified as mutually competitive.(ABSTRACT TRUNCATED AT 250 WORDS)

Diffusion↗

PubMed1993

Focal visual attention produces illusory temporal order and motion sensation.

Spatial attention was studied using a new visual illusion of motion: a line, which was presented physically at once, was perceived to be drawn from one side when attention had been captured to that side of the line by a preceding visual cue stimulus. By comparing with a temporal order task, we showed that the line-motion illusion was produced by acceleration of visual information processing at the locus of attention. The results suggest that the facilitatory effect of attention is exerted at relatively early stages of visual information processing where visual signals are to be fed into the motion detecting mechanism.

Attention↗

PubMed1993

Visual attention revealed by an illusion of motion.

Attention is a mechanism to select sensory information. It is a modulatory process which normally cannot be observed as overt responses. A new psychophysical method using an illusion of motion perception allowed us to visualize the field of the magnitude of attention and its dynamic changes. Based on our experiments using this method we suggest that (1) both passive (bottom-up) and active (top-down) attention exert their effects on the early stages of visual processing, (2) active attention can quickly and briefly be replaced by passive attention induced by an external event, but can be restored in about 400 ms, and (3) attention is directed to an object, not space, and follows the object as it moves.

Animals↗

PubMed1993

Alterations of proteoglycan synthesis in rabbit articular cartilage induced by intra-articular injection of papain.

In order to investigate the biochemical alteration of proteoglycan (PG) synthesis during cartilage repair, reversible destruction was induced by injecting papain into the knee joint cavity of rabbits. The PG synthesis in the cartilage was examined using Na2 35SO4 and high performance liquid chromatography (HPLC). PGs labeled with 35SO4(2-) (35S-PGs) were extracted from normal and papain-treated cartilage, and the amount of synthesis, ability to aggregate with hyaluronan (HA), and the composition of glycosaminoglycan and chondroitin sulfate isomer labeled with 35SO4(2-) (35S-GAG and 35S-CS isomer) were analyzed. Synthesis of 35S-PGs, especially those that were unable to aggregate with HA (nonaggregating 35S-PGs), increased in papain-treated cartilage compared with that in normal cartilage. The acceleration and qualitative change in PG synthesis in the papain-treated cartilage are considered to be responses to the supplementation of the loss of cartilage PGs induced by papain. The compositions of 35S-GAG and 35S-CS isomer of the nonaggregating 35S-PGs differed from those of 35S-PGs which were able to aggregate with HA (aggregating 35S-PGs) in the papain-treated cartilage as well as in the normal cartilage. However, the compositions of both nonaggregating and aggregating 35S-PGs in the papain-treated and normal cartilage were similar. These results indicate that most of the nonaggregating 35S-PGs in papain-treated cartilage have properties similar to those in normal cartilage and are not simple degradation products of aggregating 35S-PGs; they also suggest that the supplementary reaction for PG content in the cartilage during its repair process is not simple acceleration in PG turn-over but the enhancement of PG synthesis accompanied by alterations in aggregating ability and the compositions of GAG and CS isomer.

Animals↗

PubMed1993

Comparison of the hepatic uptake clearances of fifteen drugs with a wide range of membrane permeabilities in isolated rat hepatocytes and perfused rat livers.

The hepatic uptake clearances of 15 ligands with a wide range of permeabilities were determined in rats using two techniques: centrifugal filtration with isolated hepatocytes and the multiple indicator dilution (MID) method with isolated perfused livers. Some of the uptake clearance values were taken from the literature. Uptake clearance values obtained from isolated hepatocytes were extrapolated to that per gram liver (PSinf.cell), assuming that 1 g of liver has 1.3 x 10(8) cells. The values of PSinf.cell varied from approximately 0.1 to 72 (mL/min/g liver). The values of PSinf.cell were similar to those (PSinf.MID) determined by the MID method for ligands with uptake clearances below approximately 1 mL/min/g liver. However, for the ligands with larger uptake clearances, the PSinf.MID values were lower than the PSinf.cell values and appeared to reach an upper limit (approx. 15-20 mL/min/g liver). The PSinf.cell values of 1-propranolol, tetraphenylphosphonium (TPP+), and diazepam were 72, 43, and 22 mL/min/g liver, respectively, whereas their uptake clearances (PSinf.MID) determined by the MID method were 4 to 10 times lower. One of the possible mechanisms for this discrepancy is that an unstirred water layer, which may exist in Disse's space in isolated perfused livers (and probably under in vivo condition), limits the hepatic uptake rate of ligands with extremely high membrane permeabilities.

Animals↗

PubMed1993

Voluntary and stimulus-induced attention detected as motion sensation.

Attention may be drawn passively to a visually salient object. We may also actively direct attention to an object of interest. Do the two kinds of attention, passive and active, interact and jointly influence visual information processing at some neural level? What happens if the passive and active attentions come into conflict? These questions were addressed with the aid of a novel psychophysical technique which reveals an attentional gradient as a sensation of motion in a line which is presented instantaneously. The subjects were asked to direct attention with voluntary effort: to the side opposite to a stimulus change, to an object with a predetermined colour, and to an object moving smoothly. In every case the same motion sensation was induced in the line from the attended side to the unattended side. This voluntary attention, however, can easily and quickly be distracted by a change in the periphery, though it can be regained within a period of 200 to 500 ms. The results suggest that the line motion can be induced in voluntary (top-down) as well as stimulus-driven (bottom-up) situations, thus indicating the truly attentional nature of the effect, rather than it being some kind of retinotopic sensory artifact or response bias. The results also suggest that these two kinds of attention have facilitatory effects acting together on a relatively early stage of visual information processing.

Adult↗