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Biomedical subjects

Y Okuda

Publications and source records attributed to Y Okuda.

At least 181 records · Page 10Linked to original sources

Omega-3 polyunsaturated fatty acids inhibit migration of human vascular smooth muscle cells in vitro.

Arterial smooth muscle cell migration from the media to the intima is a crucial process in the pathogenesis of atherosclerosis. Platelet-derived growth factor (PDGF) has been proposed to play a key role in the development of advanced atherosclerotic lesions by stimulating the migration and proliferation of vascular smooth muscle cells. Polyunsaturated fatty acids (PUFA) of the omega-3 series, extracted from fish oil has been shown to have beneficial effects on atherosclerosis. In this study, we evaluated the effects of omega-3 PUFA on the migration of human aortic smooth muscle cell (hASMC) in vitro. The migration assay was performed according to the Capsoni's method using transwell culture plates. PDGF, fibrinogen or 10% FCS significantly stimulated hASMC migration, however, omega-3 PUFA significantly inhibited PDGF-induced migration of hASMC. These results suggest that the inhibitory effect of omega-3 PUFA on cell migration may be an important aspect by which omega-3 PUFA exerts its antiatherosclerotic influence.

Cell Movement↗

Acute effects of thromboxane dual blocker (KDI-792) on different portions of lower limb blood flow--a study using Doppler ultrasonography and laser Doppler flowmetry in type 2 diabetic patients.

The acute effects of a newly synthesized thromboxane dual blocker (KDI-792), a combined thromboxane synthase inhibitor and receptor antagonist, on lower limb circulation were examined using two-dimensional color and pulse Doppler ultrasonography and laser Doppler flowmetry. A randomized single-masked, placebo-controlled trial was performed on 36 type 2 diabetic patients with minimally impaired baseline flow. The anatomical cross-sectional area (CSA), maximum flow velocity (MFV) and flow volume index (FVI) in the right dorsal pedis artery (DPA) and right femoral artery (FA) were determined by Doppler ultrasonography before and 45 and 90 minutes after the administration of either 100 or 200 mg of KDI-792 to the dose groups or placebo to the control group. Periflux blood flow (PBF) in the right foot was determined simultaneously by laser Doppler flowmetry. Both CSA and MFV in the dose groups were significantly increased in both the FA and DPA. FVI was markedly increased from 21.4 +/- 3.7 to 68.3 +/- 26.8 in the DPA (M +/- SD, P < 0.01) and from 365.4 +/- 35.3 to 771.7 +/- 75.7 in the FA (P < 0.01) in the 200 mg dose group. In the 100 mg dose group, FVI was markedly increased from 20.0 +/- 8.7 to 68.3 +/- 26.8 (P < 0.01) in the DPA and from 372.5 +/- 130.0 to 677.5 +/- 187.8 (P < 0.01) in the FA. PBF was also increased in both dose groups (from 4.15 +/- 1.4 to 7.0 +/- 4.0 ml/min/100 g tissue in the 200 mg dose group, P < 0.01), whereas there were no significant changes in either measurement in the control group. There were no significant changes in pulse rate or blood pressure after administration in either the dosage group or the placebo group. These and previous findings indicate that a single administration of KDI-792 markedly increases lower limb blood flow and might have a more potent vasodilating effect than that of prostaglandin I2 derivatives.

Aged↗

Nitric oxide via an inducible isoform of nitric oxide synthase is a possible factor to eliminate inflammatory cells from the central nervous system of mice with experimental allergic encephalomyelitis.

We recently identified the inducible isoform of nitric oxide synthase (iNOS) in inflammatory lesions of the central nervous system (CNS) in mice with experimental allergic encephalomyelitis (EAE), a known animal model of multiple sclerosis (MS). In the present study, the role of excessive nitric oxide (NO) production via iNOS was investigated in mice with EAE using immunohistochemistry with antibodies to nitrotyrosine and iNOS, NADPH-diaphorase histochemistry, and the in situ terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) method to detect cell death, presumably through an apoptotic mechanism. NADPH-diaphorase histochemistry and immunohistochemistry for iNOS revealed an elevation of nitric oxide synthase (NOS) activity during the course of EAE, which came from iNOS. Nitrotyrosine was detected in infiltrated cells and some glial cells in the spinal cord lesions, where iNOS-positive inflammatory cells were present at the peak of EAE. The findings implied the generation of NO and peroxynitrite in the EAE lesions, which might damage structural and functional proteins. The TUNEL positive cells were mainly inflammatory ones, and most of them were located in close proximity to iNOS-positive cells, while some of them were iNOS-positive themselves. These results suggested that excessive NO via iNOS played an important role to eliminate inflammatory cells in the CNS of mice with EAE, possibly through an apoptotic mechanism.

Animals↗

Inhibitory effects of omega-3 polyunsaturated fatty acids on receptor-mediated non-selective cation currents in rat A7r5 vascular smooth muscle cells.

1. The effects of omega-3 polyunsaturated fatty acids on receptor-mediated non-selective cation current (Icat) and K+ current were investigated in aortic smooth muscle cells from foetal rat aorta (A7r5 cells). The whole-cell voltage clamp technique was employed. 2. With a K(+)-containing solution, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA, 30 microM) produced an outward current at a holding potential of -40 mV. This response was inhibited by tetraethylammonium (20 mM) or Cs+ in the patch pipette solution, and the reversal potential of the EPA-induced current followed the K+ equilibrium potential in a near Nernstian manner. 3. Under conditions with a Cs(+)-containing pipette solution, both vasopressin and endothelin-1 (100 nM) induced a long-lasting inward current at a holding potential of -60 mV. The reversal potential of these agonist-induced currents was about +0 mV, and was not significantly altered by the replacement of the extracellular or intracellular Cl+ concentration, suggesting that the induced current was a cation-selective current (Icat). 4. La3+ and Cd2+ (1 mM) completely abolished these agonist-induced Icat, but nifedipine (10 microM) failed to inhibit it significantly. 5. omega-3 polyunsaturated fatty acids (3-100 microM), EPA, DHA and docosapentaenoic acids (DPA), inhibited the agonist-induced Icat in a concentration-dependent manner. The potency of the inhibitory effect was EPA > DHA > DPA, and the half maximal inhibitory concentration (IC50) of EPA was about 7 microM. 6. Arachidonic and linoleic acids (10, 30 microM) showed a smaller inhibitory effect compared to omega-3 fatty acids. Also, oleic and stearic acids (30 microM) did not show a significant inhibitory effect on Icat. 7. A similar inhibitory action of EPA was observed when Icat was activated by intracellularly applied GTP gamma S in the absence of agonists, suggesting that the site of action of omega-3 fatty acids is not located on the receptor. 8. These results demonstrate that omega-3 polyunsaturated fatty acids can activate a K+ current and also effectively inhibit receptor-mediated non-selective cation currents in rat A7r5 vascular smooth muscle cells. Thus, the data suggest that omega-3 fatty acids may play an important role in the regulation of vascular tone.

Animals↗

Generation and characterization of rat monoclonal antibodies against human serum amyloid A.

Monoclonal antibodies against human serum amyloid A (SAA) were generated in rat (which seems not to have mature SAA proteins) by immunizing intact human SAA. Thirteen clones selected by initial screening were analysed based on reactivity with synthetic peptides of SAA and with carboxyl-terminal truncated recombinant SAA. Antibodies were divided into four types, i.e. those recognizing the area around residue 18, 30, 90, and 100, respectively, of SAA. The antibody to the carboxyl terminus (around residue 100) of SAA, when subjected to immunohistochemistry for amyloid deposits in specimens from patients with reactive amyloidosis, always yielded negative reactivity, supporting the general concept that the carboxyl terminus of SAA is absent from human AA deposits.

Amino Acid Sequence↗

Comparison of stellate ganglion block with intravascular infusion of prostaglandin E1 on brachial artery blood flow in dogs.

We sought to determine whether sympathetic blockade or infusion of prostaglandin E1 (PGE1) is better for vasodilation. We measured brachial artery blood flow (BABF) in 10 mongrel dogs using an ultrasonic time flowmeter to compare the effects of stellate ganglion block (SGB) and intravascular infusion of PGE1. The experimental protocol was designed as follows: 1) intravenous (IV) infusion of PGE1 at a rate of 10 ng x kg(-1) x min(-1) for 10 min, 2) IV infusion of PGE1 at a rate of 150 ng x kg(-1) x min(-1) for 10 min, 3) intraarterial infusion of PGE1 at a rate of 0.1 ng x kg(-1) x min(-1) for 10 min, 4) SGB with 0.5% mepivacaine 1.0 mL was used as a sympathetic blockade. These procedures were successively performed on each dog. Mean arterial pressure (MAP), heart rate (HR), and BABF were measured before and after each procedure for 40 min. MAP and HR did not change significantly after the procedures. BABF increased significantly after IV infusion of PGE1 150 ng x kg(-1) x min(-1), intraarterial infusion of PGE1 and SGB, reaching maximums of 157%, 174%, and 171% 10 min after IV infusion of PGE1 150 ng x kg(-1) x min(-1), intraarterial infusion of PGE1 and SGB compared with the prevalues, respectively. These data indicate that sympathetic blockade may produce the same vasodilation as IV infusion of PGE1 150 ng x kg(-1) x min(-1) and intraarterial infusion of PGE1 0.1 ng x kg(-1) x min(-1). Intravascular infusion of PGE1 could provide clinically equivalent vasodilation without the complications associated with SGB.

Alprostadil↗

Intractable diarrhoea associated with secondary amyloidosis in rheumatoid arthritis.

OBJECTIVE: To examine the clinical characteristics of intractable diarrhoea associated with secondary amyloidosis in rheumatoid arthritis (RA). METHODS: Of 179 RA patients with biopsy confirmed secondary amyloidosis, 24 cases (23 women and one man) with intractable diarrhoea lasting for more than one month were retrospectively evaluated. RESULTS: The mean (SD) duration of diarrhoea was 87 (64) days. Prodromal symptoms of gastrointestinal dysfunction (n = 21) and impaired peristalsis (n = 16) were observed. Laboratory data showed hypoproteinaemia (4.7 (0.85) g/dl) caused by malabsorption or protein loss and high values of C reactive protein (17.0 (9.3) mg/dl). Recurrence of intractable diarrhoea (n = 4) and transition from intractable diarrhoea to other gastrointestinal problems of amyloidosis (ischaemic colitis (n = 2) and intestinal pseudo-obstruction (n = 4)) were observed. In 19 patients (25 episodes) the duration of intravenous hyperalimentation at remission (18 episodes) was 68 (52) days. Corticosteroid pulse therapy was administered to 10 patients (11 times) and the time elapsed from the end of corticosteroid pulse therapy to the end of diarrhoea was 18 (14) days. One and five year survival rates after the onset of intractable diarrhoea were 73.4% and 38.9%. Seven of 13 patients (54%) had died as a result of infectious diseases. CONCLUSION: Intractable diarrhoea associated with secondary amyloidosis in RA is a serious clinical entity and the prognosis is poor. Although it is assumed that intravenous hyperalimentation treatment and corticosteroid pulse therapy are favourable regimens for intractable diarrhoea, the patients should be monitored for possible infectious complications.

Adult↗

Is mitomycin effective in preventing muscle migration after hang-back recession in a rabbit model?

To investigate the effect of intraoperative mitomycin C (MMC) application on muscle insertion migration after adjustable surgery, hang-back recession was bilaterally performed for 4 mm on the inferior rectus muscle in 30 rabbits. The scleral wound of the insertion site of one eye of each pair was treated with a sponge soaked with a solution cotaining mitomycin at a concentration of either 0.4 or 1.0 mg/ml for 5 min. As a control, the contralateral eye was treated with a distilled-water-soaked sponge. Three, 6 and 12 weeks later, the distance from the anterior border of the reattached muscle to the original insertion was measured with in vivo and microscopic examination. Anterior migration of the muscle insertion was observed in both the MMC-treated and control eye. The measurement by the in vivo and histologic examinations revealed that MMC treatment did not significantly reduce the anterior migration relative to the control eye. Analysis of the time course in the MMC-treated eye revealed a significant increase during the observation period in anterior migration as measured by the histologic examination (p = 0.0267 for the 0.4 mg/ml group and p = 0.0408 for the 1.0 mg/ml group). Exposure to MMC has no significant inhibitory effect on the muscle migration compared to that in the control eyes.

Animals↗

Nitric oxide production by cultured rat Leydig cells.

Nitric oxide (NO) has emerged as an intracellular and intercellular messenger in a number of biological systems. In the present study, we demonstrated that NO was produced by cultured rat Leydig cells, and that inducible nitric oxide synthase (iNOS) messenger RNA (mRNA) was expressed in Leydig cells. NO was measured as nitrite with the method of Griess. Although unstimulated Leydig cells produce little NO, interleukin-1 beta (IL-1 beta) markedly increased NO production. NO production was inhibited by the NOS inhibitor, NG-monomethyl-L-arginine. Northern blot analysis showed that iNOS mRNA was little expressed in freshly isolated immature Leydig cells, but that iNOS mRNA levels were increased by the addition of IL-1 beta in a dose-dependent manner at the concentration up to 10 ng/ml. The levels of iNOS mRNA were increased as early as 3 h after the addition of IL-1 beta and persisted for up to 24 h. In adult Leydig cells, IL-1 beta stimulated iNOS mRNA expression. Immunocytochemical analysis demonstrated iNOS-like immunoreactivity in the cytoplasm of Leydig cells. These results indicate that NO is produced in Leydig cells and suggest that NO might be involved in the physiological function of Leydig cells.

Animals↗

[Chondroblastoma of the temporal bone: a case report].

A 30-year-old male had been suffering from left temporalgia of six months duration and then developed left hearing disturbance. Craniogram and bone window CT revealed a well defined osteolytic lesion in the left temporal bone. CT scan showed an expansile heterogenous mass with calcification. Both T1 and T2 weighted MRI demonstrated a well lobulated mixed intensity mass, but no evidence of dural or intracranial invasion. The tumor exhibited homogenous enhancement on CT and MRI. Angiogram revealed a well marked staining supplied by the left middle meningeal and deep temporal arteries. Subtotal removal of the tumor was carried out with cranioplasty. Histologically, this tumor was composed of round or polygonal chondroblasts, scattered osteoclast-like giant cells with a foci of cartilage in the stroma. Many reports describe giant cell tumor can be differentiated by immunohistochemical demonstration of S100 protein. Although in our case, histological findings simulated those of eosinophilic granuloma, it was diagnosed as chondroblastoma because of the foci of cartilage in the stroma. Because this tumor is usually benign, recurrence of the tumor is rare after surgical resection. Post-operative irradiation has been reported to be effective in decreasing the recurrence of the tumor. But it should be carefully observed because of possible sarcomatous change in such tumors.

Adult↗

[Where is a leak point detected by "the low flow leak test" of anesthetic machines?].

"The low flow leak test" is recommended for pre-anesthetic inspection of anesthetic machines. We carried out anesthesia compression tests as a standard. Even in that case, often the low flow leak test does not meet the standard. We investigated the point where there is a leak in the anesthetic machine. Observing the leak that fluctuates each time there is detachment or attachment of the canister, the primary cause of the leak is thought to be related to the canister. It is important to carry out an inspection of the canister if the low flow leak test does not meet the standard.

Anesthesiology↗

Cloning and sequencing of a gene encoding a new member of the tetratricopeptide protein family from magnetosomes of Magnetospirillum magnetotacticum.

A gene encoding the 22 kDa protein (MAM22) which was localized in the magnetosomes isolated from the magnetotactic bacterium, Magnetospirillum magnetotacticum, was cloned and sequenced. MAM22 was composed of 220 amino acids (aa) with a molecular weight of 24,186 Da. The deduced aa sequence exhibited significant homology with a number of proteins that belong to the tetratricopeptide repeat (TPR) protein family, including mitochondrial protein import receptors and peroxisomal protein import receptors. The presence of three repeats of a degenerate 34-aa consensus sequence, suggest that MAM22 localized in magnetosome membranes may interact with the cytoplasmic proteins containing similar TPR motifs.

Amino Acid Sequence↗