Search PubMed⌕ Search

Biomedical subjects

T Ohhashi

Publications and source records attributed to T Ohhashi.

At least 19 recordsLinked to original sources
PubMed2001

Effects of diets enriched in n-6 or n-3 fatty acids on cholesterol metabolism in older rats chronically fed a cholesterol-enriched diet.

Hypocholesterolemic effects in older animals after long-term feeding are unknown. Therefore, aged rats (24 wk of age) fed a conventional diet were shifted to diets containing 10% perilla oil [PEO; oleic acid + linoleic acid + alpha-linolenic acid; n-6/n-3, 0.3; polyunsaturated fatty acid/saturated fatty acid (P/S), 9.6], borage oil [oleic acid + linoleic acid + alpha-linolenic acid; n-6/n-3, 15.1; P/S, 5.3], evening primrose oil (EPO; linoleic acid + gamma-linolenic acid; P/S, 10.5), mixed oil (MIO; oleic acid + linoleic acid + gamma-linolenic acid + alpha-linolenic acid; n-6/n-3, 1.7; P/S, 6.7), or palm oil (PLO; palmitic acid + oleic acid + linoleic acid; n-6/n-3, 25.3; P/S, 0.2) with 0.5% cholesterol for 15 wk in this experiment. There were no significant differences in the food intake and body weight gain among the groups. The liver weight in the PEO (n-6/n-3, 0.3) group was significantly higher than those of other groups in aged rats. The serum total cholesterol and very low density lipoprotein (VLDL) + intermediate density lipoprotein (IDL) + low density lipoprotein (LDL)-cholesterol concentrations of the PLO (25.3) group were consistently higher than those in the other groups. The serum high density lipoprotein cholesterol concentrations of the PEO (0.3) and EPO groups were significantly lower than in the other groups at the end of the 15-wk feeding period. The liver cholesterol concentration of the PLO (25.3) group was significantly higher than those of other groups. There were no significant differences in the hepatic LDL receptor mRNA level among the groups. Hepatic apolipoprotein (apo) B mRNA levels were not affected by the experimental conditions. The fecal neutral steroid excretion of the PLO (25.3) group tended to be low compared to the other groups. The results of this study demonstrate that both n-6 fatty acid and n-3 fatty acids such as gamma-linolenic acid and alpha-linolenic acid inhibit the increase of serum total cholesterol and VLDL + IDL + LDL-cholesterol concentrations of aged rats in the presence of excess cholesterol in the diet compared with dietary saturated fatty acid.

Aging↗
PubMed2001

B16-BL6 melanoma cells release inhibitory factor(s) of active pump activity in isolated lymph vessels.

We investigated whether supernatant cultured with melanoma cell lines B16-BL6 and K1735 or the Lewis lung carcinoma cell line (LLC) can regulate lymphatic pump activity with bioassay preparations isolated from murine iliac lymph vessels. B16-BL6 and LLC supernatants caused significant dilation of lymph microvessels with cessation of pump activity. B16-BL6 supernatant produced dose-related cessation of lymphatic pump activity. There was no significant tachyphylaxis in the supernatant-mediated inhibitory response of lymphatic pump activity. Pretreatment with 3 x 10(-5) M N(omega)-nitro-L-arginine methyl ester (L-NAME) or 10(-7) M or 10(-6) M glibenclamide and 5 x 10(-4) M 5-hydroxydecanoic acid caused significant reduction of supernatant-mediated inhibitory responses. Simultaneous treatment with 10(-3) M L-arginine and 3 x 10(-5) M L-NAME significantly lessened L-NAME-induced inhibition of the supernatant-mediated response, suggesting that endogenous nitric oxide (NO) plays important roles in supernatant-mediated inhibitory responses. Chemical treatment dialyzed substances of <1,000 molecular weight (MW), producing complete reduction of the supernatant-mediated response. In contrast, pretreatment with heating or digestion with protease had no significant effect on supernatant-mediated response. These findings suggest that B16-BL6 cells may release nonpeptide substance(s) of <1,000 MW, resulting in significant cessation of lymphatic pump activity via production and release of endogenous NO and activation of mitochondrial ATP-sensitive K(+) channels.

Animals↗
PubMed2001

Parathyroid hormone-related protein-(1-34) inhibits intrinsic pump activity of isolated murine lymph vessels.

Parathyroid hormone-related protein (PTHrP) was originally found as a tumor-derived vasoactive factor and has also been known to produce significant relaxation of vascular smooth muscles. Thus effects of PTHrP-(1-34), a PTH receptor-binding domain, on spontaneous lymphatic pump activity was investigated in isolated pressurized lymph vessels of mice. Low concentrations (1 x 10(-10) and 3 x 10(-10) M) of PTHrP-(1-34) dilated lymph vessels and reduced the frequency of pump activity, whereas high concentrations (1 x 10(-9) to 1 x 10(-8) M) of PTHrP-(1-34) caused dilation with cessation of the lymphatic pump activity. N(omega)-nitro-L-arginine methyl ester (L-NAME; 3 x 10(-5) M) but not indomethacin (1 x 10(-5) M) significantly reduced the PTHrP-(1-34)-induced inhibitory responses of the lymphatic pump activity. In the presence of L-NAME (3 x 10(-5) M) and L-arginine (1 x 10(-3) M), the L-NAME-induced inhibition in the PTHrP-(1-34)-mediated responses was significantly reduced. Glibenclamide (1 x 10(-6) M) significantly suppressed the inhibitory responses of the lymphatic pump activity induced by PTHrP-(1-34) and S-nitroso-N-acetyl-penicillamine. The PTHrP-(1-34)-mediated inhibitory responses were significantly reduced by treatment with PTHrP-(7-34) (1 x 10(-7) M). These results suggest that PTHrP-(1-34) inhibits spontaneous pump activity of the isolated lymph vessels via PTH receptors and that production and release of endogenous nitric oxide and activation of ATP-sensitive K(+) channels in the lymph vessels contribute to the PTHrP-(1-34)-mediated inhibitory responses of the lymphatic pump activity.

Animals↗
PubMed2001

[Lymphodynamic factors governing lymphatic spread of carcinoma cells].

Carcinoma frequently spreads and grows through the lymphatic system, whereas malignant tumors of mesenchymal origin more frequently spread via the hematogenous route. This paper reviews cancer spread with special reference to tumor microcirculation, vascular endothelial growth factor receptor (VEGFR)-3-mediated lymphangiogenesis, regeneration of collecting lymph vessels, and the role of nitric oxide in lymphatic metastasis. Oxygen and pH are key microenvironmental factors in the development and growth of tumors and their response to treatment. Thus the physiological and pathophysiological roles of hypoxia and acidic pH in the tumor microenvironment in lymphatic metastasis are demonstrated. Lymph angiogenesis in tumor tissues and regeneration of collecting lymph vessels with special reference to VEGFR-3 and its agonists VEGF-C/VEGF-D are discussed. The occurrence and biological significance of intratumoral lymph angiogenesis in breast cancer have been established. VEGF-C as a molecular link between tumor lymph angiogenesis and metastasis is identified. Finally, the role of nitric oxide (NO) in tumor microcirculation and lymphatic metastasis of carcinoma cells is evaluated.

Humans↗
PubMed2000

Physiological roles of endogenous nitric oxide in lymphatic pump activity of rat mesentery in vivo.

Physiological roles of endogenous nitric oxide (NO) in the lymphatic pump activity of rat mesenteries in vivo were evaluated using an intravital video microscope system. Changes in the pumping frequency (F), the end diastolic diameter (EDD), and the end systolic diameter (ESD) of the mesenteric lymph microvessels were measured with the microscope system and then the pump flow index (PFI) was calculated. A 15-min superfusion of 30 microM N(omega)-nitro-L-arginine methyl ester (L-NAME) in the mesenteries caused significant increases of F and PFI and a significant decrease of the EDD and ESD. Simultaneous superfusion of 1 mM L-arginine with 30 microM L-NAME produced a significant reversal of the L-NAME-mediated increase of F and decrease of ESD. A 15-min superfusion of 100 microM aminoguanidine caused no significant effects on F, EDD, and ESD of the mesenteric lymph vessels in vivo. These findings suggest that endogenous NO has physiologically modulated the lymphatic pump activity in rat mesentery in vivo and that the production and release of NO may be mediated by constitutive NO synthase but not by inducible NO synthase.

Animals↗
PubMed2000

Development of an experimental apparatus for investigating lymphatic pumping activity of murine mesentery in vivo.

The present study has been attempted to establish a modified intravital microscope system for investigating murine lymphatic pumping activity in vivo and evaluate whether or not there is rhythmic pumping activity of murine mesenteric lymphatic vessels in vivo. We designed and constructed a custom organ chamber with a semicircular channel (8 mm in radius, 5 mm in width, 3 mm in depth), being suitable for the superfusing of murine mesentery in vivo. A marked lymphatic pumping activity was observed in the mesenteries of DDY mice. The maximal and minimal diameter and frequency in the pumping activity were 60.9 +/- 1.0 microm, 53.7 +/- 1.8 microm and 12.8 min(-1) (n = 5), respectively. Both NE (norepinephrine, 10(-8)-10(-6) M) and TEA (tetraethylammonium, 1-10 mM) caused dose-dependent constriction of the mesenteric lymphatic vessels in the mice. These findings suggest that a modified intravital microscope system with a specially designed and constructed edge-monitoring device enables us to investigate in vivo lymphatic circulation in murine mesenteries.

Animals↗
PubMed2000

Effects of VEGF on Ca(2+)-transient in cultured lymphatic endothelial cells and mechanical activity of isolated lymph vessels.

We investigated the effects of vascular endothelial growth factor (VEGF(165)) on [Ca(2+)](i)-transient in cultured lymphatic endothelial cells (LEC) and mechanical activity of isolated dog thoracic ducts. VEGF (0.1-10 ng/ml) caused a dose-dependent increase of the [Ca(2+)](i) in LEC. Pretreatment with 10(-5) M genistein or 5x10(-6) M herbimycin A produced a significant reduction of the VEGF-induced [Ca(2+)](i)-transient. In the presence of 10(-6) M thapsigargin, VEGF caused no significant effect on the [Ca(2+)](i)-transient. Pretreatment with Ca(2+)-free solution containing 0.1 mM EGTA produced no significant effect on the peak increase of [Ca(2+)](i) induced by 0.1 or 10 ng/ml VEGF, but significantly depressed the sustained part of [Ca(2+)](i) observed at the higher concentration of VEGF. The VEGF (0.1-10 ng/ml) caused a significant dilation of the isolated lymph vessels with intact endothelium, which were precontracted with U46,619. The 10 ng/ml VEGF-induced dilation was significantly reduced by 3 x 10(-5) M N(omega)-nitro-L-arginine methyl ester (L-NAME). The action of L-NAME was inhibited by the simultaneous application of 10(-3) M L-arginine. Mechanical rubbing of the endothelium also caused significant inhibition of the VEGF-induced dilation. The findings suggest that VEGF(165) may activate the receptor-related tyrosine kinase and cause the release of Ca(2+) from the inositol 1,4, 5-triphosphate-sensitive intracellular Ca(2+) stores in LEC. VEGF(165) also produces endothelium-dependent nitric oxide-mediated dilation of the precontracted isolated lymph vessels.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
PubMed2000

Effects of leg rotation on lymph flow and pressure in rabbit lumbar lymph circulation: in vivo experiments and graphical analysis.

The intraluminal lymphatic pressure in rabbit lumbar lymphatic trunks was determined. To estimate the lymphatic pressure, graphical analysis was performed from lymph outflow pressure-flow rate relationship and lymph infusion pressure-flow rate relationship. A direct measurement technique with a T-tube was also carried out to measure the pressure in the lumbar lymphatic. The rabbit leg was passively rotated at 0.3 Hz in the diameter of 8 cm to enhance the lymphatic pump activity of the leg. The estimated pressure and measured pressure in the lymphatics were 6.50 and 7.14 +/- 2.38 cmH2O, respectively. It was confirmed that similar values could be obtained from these two methods. The lymphatic pumping in the leg may affect a distribution of pressure in the lumbar lymphatic system.

Animals↗
PubMed1999

Effects of endothelin on spontaneous contractions in lymph vessels.

A mode of action of endothelin (ET) on spontaneous contractions was investigated in ring preparations of isolated bovine mesenteric lymphatics. ET-1 at concentrations between 10(-10) and 10(-9) M caused a dose-dependent increase in the frequency of spontaneous contractions. The specific ET(A)-receptor antagonist BQ-123 (5 x 10(-7) M) caused a significant inhibition of the ET-1-induced positive chronotropic effect in the ring preparations with and without the endothelium. Mechanical denudation of the lymphatic endothelial cells produced a significant potentiation of the ET-induced positive chronotropic effect. BQ-3020 (10(-8)-10(-7) M), a selective ET(B)-receptor agonist, induced dose dependently negative chronotropic and inotropic effects on the spontaneous contractions in the ring preparations with intact endothelium. Mechanical removal of the endothelium caused a significant reduction of the BQ-3020-induced negative chronotropic and inotropic effects. The ET-1-induced positive chronotropic effect was potentiated by pretreatment with N(omega)-nitro-L-arginine methyl ester (L-NAME) (10(-5) M) but unaffected by aspirin (10(-5) M). Additional treatment with L-arginine (10(-4) M) completely reversed the L-NAME-mediated potentiation of the ET-induced chronotropic effect. These results suggest that stimulation of ET(A) receptors on the lymphatic smooth muscles causes a positive chronotropic effect on the spontaneous contractions, and stimulation of ET(B) receptors on the lymphatic endothelial cells induces a release of nitric oxide, which results in the chronotropic and inotropic effects on spontaneous contractions in isolated bovine mesenteric lymphatics.

Animals↗
PubMed1999

Involvement of ATP-sensitive K(+) channels in spontaneous activity of isolated lymph microvessels in rats.

Physiological roles of ATP-sensitive K(+) channels for spontaneous activity in isolated rat mesenteric lymph microvessels (maximum diameter approximately 80-150 microm) were investigated. The lymph microvessels were cannulated with glass micropipettes and pressurized at a perfusion pressure of 6 cmH(2)O. Changes in the diameter and frequency of spontaneous contractions in the lymphatics were measured with videomicroscopy. Pinacidil (K(+)-channel opener) inhibited the spontaneous activity. In the presence of glibenclamide (selective ATP-sensitive K(+)-channel blocker; 10(-7) and 10(-6) M) and tetraethylammonium (TEA; nonselective K(+)-channel blocker; 10(-4) and 10(-3) M), the pinacidil-induced inhibition of the spontaneous contractions in lymph microvessels was significantly reversed. Glibenclamide and TEA themselves, however, did not affect the frequency of spontaneous activity in the lymph microvessels. These results suggest that ATP-sensitive K(+) channels are involved in the regulation of spontaneous activity in the smooth muscles of isolated lymph microvessels of rat mesenteries.

Adenosine Triphosphate↗
PubMed1998

Electrical stimulation-induced alpha1- and alpha2-adrenoceptors-mediated contraction in isolated dog thoracic ducts.

The electrical stimulation-induced responses of isolated dog thoracic ducts were investigated using an organ bath technique. Electrical stimulation (0.7 ms in pulse width, 25 V in nominal voltage, 10 s in duration time, 1-32 Hz at frequency) produced frequency-related contractions in the lymphatic preparations. The contractions were abolished by pretreatment with tetrodotoxin (10(-7) M), guanethidine (10(-7), 10(-6) M), and bretylium (10(-7), 10(-6) M). Cocaine (10(-6) M) significantly potentiated the electrical stimulation-induced contractions. Phentolamine (10(-8)-10(-5) M), prazosin (10(-8)-10(-5) M), bunazosin (10(-6), 10(-5) M), yohimbine (10(-8)-10(-6) M) and rauwolscine (10(-8)-10(-6) M) also dose-dependently reduced the contractions. On the other hand, propranolol (10(-8)-10(-6) M), atropine (10(-6) M), hexamethonium (10(-6) M), aspirin (3 x 10(-5) M), N(omega)-nitro-L-arginine methyl ester (L-NAME) (3 x 10(-5) M) and L-NAME (3 x 10(-5) M) + L-arginine (10(-4) M) caused no significant effect on electrical stimulation-induced contractions. No significant difference in the electrical stimulation-induced responses was observed between the lymphatic preparations with and without an intact endothelium. The electrical stimulation caused only a small contraction with no relaxation in the thoracic duct preparation precontracted with 10(-8) M U46619. The small contraction was abolished by 10(-5) M phentolamine. These findings suggest that there exists alpha1- and alpha2-adrenoceptors-mediated excitatory innervation, but no NO-ergic inhibitory nerve fiber in dog thoracic ducts.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
PubMed1998

Human perspiration measurement.

We review various methods developed for human perspiration measurement and their physiological applications, with special reference to the performance and application of a new home-made ratemeter and instrumentation with a microscope. Many kinds of humidity sensor based on humidity-sensitive electrical properties have been investigated and placed on the market. Recently a capacitive thin-film humidity sensor was constructed and confirmed to be one of the best humidity sensors for accurately and quickly detecting changes in the relative humidity of gas-flow perfused through a ventilated chamber for human perspiration measurement. In this paper we also introduce a new home-made ratemeter with a capacitive humidity sensor, the electrical output of which is not disturbed by changes in ambient temperature, and new instrumentation for directly observing drops of sweat secreted from eccrine glands in human skin and simultaneously measuring the change in amount of perspiration at the same area of skin. Finally, we review physiological applications of the methods for measuring human palmar perspiration including emotional sweating.

Humans↗
PubMed1997

Capsaicin-induced nitric-oxide-dependent relaxation in isolated dog urethra.

Capsaicin (5 x 10[-8] to 5 x 10[-5] M) produced a non-adrenergic and non-cholinergic phasic relaxation in a concentration-dependent manner in isolated dog urethral preparations precontracted by noradrenaline. The mode of action of capsaicin was investigated with special reference to the possible involvement of endogenous nitric oxide (NO). A marked tachyphylaxis was observed in the responses to capsaicin. Pretreatment with NG-nitro-L-arginine-methyl-ester (L-NAME) prevented or markedly reduced the inhibitory effect of L-NAME. Methylene blue inhibited the capsaicin-induced relaxation. In preparations stored at 4 degrees C for 72 h, the reduction in the capsaicin-induced relaxation was significantly greater than that in the relaxation induced by either electrical field stimulation or by sodium nitroprusside. We conclude that capsaicin produces an endogenous-NO-dependent relaxation in the isolated dog urethra via mechanisms that deteriorate during cold storage of the preparations.

Animals↗
PubMed1997

The primary structure of the Cl(-)-translocating ATPase, b subunit of Acetabularia acetabulum, which belongs to the F-type ATPase family.

The genes possibly encoding the b subunit (50 kDa) of the Cl(-)-translocating ATPase of Acetabularia acetabulum were cloned from total RNA and from poly(A)+ RNA and sequenced. The deduced amino acid sequence of the open reading frame consisted of 478 amino acids and showed high similarity to the beta subunit of chloroplast F1-ATPase. Gene fragments encoding the putative beta subunit of chloroplast F1- (273 bp) and mitochondrial F1-ATPases (332 bp) were also cloned from A. acetabulum and sequenced, respectively. The deduced amino acid sequence of the chloroplast F1-ATPase showed 92.5% identity to be primary structure of the b subunit of the Cl(-)-translocating ATPase, while the nucleotide sequences were 79.9% identical. The deduced amino acid sequence of the latter was 77.3% identical to that of the b subunit of the Cl(-)-translocating ATPase and the nucleotide sequences were 67.5% identical. By Northern analysis, these three beta-like genes were demonstrated to be transcribed with different sizes of RNA species. A putative chloroplast F1-beta fragment also hybridized with chloroplast DNA isolated from the organism.

Acetabularia↗
PubMed1997

Inhibitory effects of fluorescein isothiocyanate photoactivation on lymphatic pump activity.

The effects of photoactivation of fluorescein 5'-isothiocyanate (FITC)-dextran on lymphatic pump activity of rat mesenteric collecting vessel were studied in vivo. Rats were anesthetized with intraperitoneal alpha-chloralose and urethane, and the mesenteries were studied by using intravital videomicroscopic techniques. The diameter of the collecting lymph vessels were continuously monitored and lymphatic pump parameters (end diastolic diameter, end systolic diameter, stroke volume index, ejection fraction, contraction frequency, and pump flow index) were calculated. FITC-dextran (42 nmol/100 g body wt) without illumination caused no disturbance of lymphatic pump activity. Photoactivated FITC-dextran significantly increased end systolic diameter and decreased stroke volume index, ejection fraction, contraction frequency, and pump flow index. End diastolic diameter was not changed throughout the experiment. Superoxide dismutase (120 U/ml) and catalase (5000 U/ml) had no protective effect on photoactivated FITC-induced pump dysfunction, while histidine (singlet oxygen quencher, 10 mM) significantly prevented the disturbance of pump parameters. These results indicate that photoactivation of FITC induces negative chronotropic and negative inotropic effects in lymphatic pump activity through generation of singlet oxygen in the mesentery.

Animals↗
PubMed1997

Instrumentation of a handy microscopic probe for concurrent observation and measurement of active sweat secretion, and its applications.

Instrumentation for the concurrent, dynamic monitoring of active sweat glands and perspiration volume is described. A device for the measurement of the rate of sweat secretion was installed on the head part of a microscope. The combined apparatus (microscopic probe) is handy for use and its weight is very light (ca. 300 g). The microscopic probe is easily attached to the surface of human skin. The dynamic activities of the sweat glands on the forehead and nose and under the nose were observed and measurement when thermal, mental and physical stimuli were applied. The activities of individuals sweat glands were asynchronous when observed in units of a few seconds or less; however, they worked synchronously in a unit period of several seconds. The latter were recorded as fine peaks by a strip chart recorder. The proposed system may be useful for the study of the sympathetic nervous system, the skin sympathetic reflex and the working of sudomotor nerves.

Adult↗
PubMed1997

Flow- and agonist-mediated nitric oxide- and prostaglandin-dependent dilation in spinal arteries.

Isolated rabbit spinal resistance-sized arteries (approximately 100 microns in diameter and approximately 3 mm long) were cannulated at both ends with glass micropipettes and perfused at constant pressure (60 mmHg). An increase of flow rate corresponding to a change of pressure gradient (delta P) ranging from 0 to 20 mmHg produced a flow-dependent vasodilation. Treatment with 50 microM aspirin or 10 microM indomethacin produced a significant reduction of the flow-dependent vasodilation only at delta P of 5 mmHg. In contrast, treatment with N omega-nitro-L-arginine methyl ester (L-NAME, 30 microM) produced no significant change. In the presence of 10 microM indomethacin, however, 30 microM L-NAME caused a marked decrease in the arterial diameter at delta P of 5 mmHg, which was completely reversed with additional administration of 1 mM L-arginine. Acetylcholine (ACh) produced a dose-dependent increase in the arterial diameter. The ACh-induced vasodilation was significantly reduced by 10 microM indomethacin or 50 microM aspirin and partially suppressed by 30 microM L-NAME. Pretreatment with both indomethacin and L-NAME completely reduced the ACh-induced vasodilation. In the presence of 10 microM indomethacin, additional treatment with 1 mM L-arginine significantly reversed the L-NAME-induced inhibition of the ACh-mediated vasodilation. Endothelial removal with Triton X-100 significantly reduced the ACh-induced vasodilation. Isocarbacyclin (a stable prostaglandin I2 analogue), prostaglandin E2, and arachidonic acid caused a dose-dependent dilation in the small arteries. These findings suggest that prostanoids play a major role in the flow- or ACh-induced vasodilation in the rabbit spinal resistance-sized small arteries.

Acetylcholine↗