Biomedical subjects

A Ishimatsu

Publications and source records attributed to A Ishimatsu.

18 recordsLinked to original sources

PubMed2003

Acute CO2 tolerance during the early developmental stages of four marine teleosts.

Ocean sequestration of CO2 is proposed as a possible measure to mitigate climate changes caused by increasing atmospheric concentrations of the gas, but its impact on the marine ecosystem is unknown. We investigated the acute lethal effect of CO2 during the early developmental stages of four marine teleosts: red sea bream (Pagrus major), Japanese whiting (Sillago japonica), Japanese flounder (Paralichthys olivaceus), and eastern little tuna (Euthynnus affinis). The percentages of larvae that hatched and survived were not affected by exposure to water with a PCO2 of 1.0 kPa (= 7.5 mmHg) within 24 h. Median lethal PCO2 values for a 360-min exposure were 1.4 kPa (cleavage), 5.1 kPa (embryo), 7.3 kPa (preflexion), 4.2 kPa (flexion), 4.6 kPa (postflexion), and 2.5 kPa (juvenile) for red sea bream; 2.4 kPa (cleavage), 4.9 kPa (embryo), 5.9 kPa (preflexion), 6.1 kPa (flexion), 4.1 kPa (postflexion), and 2.7 kPa (juvenile) for Japanese whiting; 2.8 kPa (cleavage) and > 7.0 kPa (young) for Japanese flounder; and 11.8 kPa (cleavage) for eastern little tuna. Red sea bream and Japanese whiting of all ontogenetic stages had similar susceptibilities to CO2: the most susceptible stages were cleavage and juvenile, whereas the most tolerant stages were preflexion and flexion.

Animals↗

PubMed2000

Mechanism of superoxide anion generation in the toxic red tide phytoplankton Chattonella marina: possible involvement of NAD(P)H oxidase.

Red tide phytoplankton Chattonella marina is known to produce reactive oxygen species (ROS), such as superoxide anion (O(2)(-)), hydrogen peroxide (H(2)O(2)) and hydroxyl radical (&z.rad;OH), under normal physiological conditions. Although several lines of evidence suggest that ROS are involved in the mortality of fish exposed to C. marina, the mechanism of ROS generation in C. marina remains to be clarified. In this study, we found that the cell-free supernatant prepared from C. marina cells showed NAD(P)H-dependent O(2)(-) generation, and this response was inhibited by diphenyleneiodonium, an inhibitor of mammalian NADPH oxidase. When the cell-free supernatant of C. marina was analyzed by immunoblotting using antibody raised against the human neutrophil cytochrome b558 large subunit (gp91phox), a main band of approximately 110 kDa was detected. The cell surface localization of the epitope recognized with this antibody was also demonstrated in C. marina by indirect immunofluorescence. Furthermore, Southern blot analysis performed on genomic DNA of C. marina with a probe covering the C-terminal region of gp91phox suggested the presence of a single-copy gene coding for gp91phox homologous protein in C. marina. These results provide evidence for the involvement of an enzymatic system analogous to the neutrophil NADPH oxidase as a source of O(2)(-) production in C. marina.

Amino Acid Sequence↗

PubMed2000

Aerial ventilatory responses of the mudskipper, Periophthalmodon schlosseri, to altered aerial and aquatic respiratory gas concentrations.

Periophthalmodon schlosseri is a mudskipper which uses the vascularized buccopharyngeal cavity as a respiratory organ. The fish construct mud burrows that contain hypoxic water, but store air inside the burrows. Because the burrow gas is frequently hypoxic and hypercapnic, the effects of altered respiratory gas concentrations on the aerial ventilation frequency (V(F)), inspiratory tidal volume (V(T)) and minute volume (V(M)=V(F)xV(T)) of P. schlosseri were studied by pneumotachography. Both total buccopharyngeal gas volume (V(BP)) and V(T) scaled significantly with body mass (mass exponents=1.10 and 1.03, respectively), and V(T)/V(BP) was 0.54+/-0. 05 (S.E.M., n=6). V(BP), expressed as a percentage of body volume, was much higher (16%) than in other air-breathing gobies (2-4%). When fish respired in normoxic air and water, V(F) was 0.25+/-0.04 breaths min(-1), V(T) 7.6+/-0.6 ml 100 g(-1), and V(M) 1.80+/-0.18 ml 100 g(-1) min(-1). Aquatic hypoxia did not significantly affect V(F), V(T), or V(M). In both moderate (P(O(2))=10 kPa) and severe (P(O(2))=5 kPa) aerial hypoxia, V(F) and V(M) increased significantly. V(T) increased significantly only during severe aerial hypoxia. In aerial hypercapnia, V(F) and V(M) increased significantly.

Amphibians↗

PubMed2000

Concanavalin A-induced discharge of glycocalyx of raphidophycean flagellates, Chattonella marina and Heterosigma akashiwo.

Chattonella marina and Heterosigma akashiwo, known as red tide phytoplankton, are naturally wall-less and have quite fragile cell structures. In this study, we found that an equilibrium dialysis technique allowed the study of lectin binding to these flagellates. The results suggested that concanavalin A (Con A) binds to these flagellate cells through the specific carbohydrate moieties on the cell surface. Interestingly, the binding of an excess of Con A on the cell surface caused morphological changes concomitant with discharge of glycocalyx, a polysaccharide-containing common structure on the external cell surface of these flagellates. Fluorescent microscopic observation using FITC-labeled Con A (F-Con A) confirmed that F-Con A molecules are localized on the discharged glycocalyx.

Animals↗

PubMed2000

Galacturonic-acid-induced increase of superoxide production in red tide phytoplankton Chattonella marina and Heterosigma akashiwo.

Red tide phytoplankton, Chattonella marina and Heterosigma akashiwo, are known to generate superoxide anion (O2-). We found that galacturonic acid (GaLUA) stimulated C. marina and H. akashiwo to generate increased amounts of O2-. Since such effect was not observed in any other monosaccharides tested, our results suggest that the binding of GalUA to specific sites on the flagellate cell surface may induce the increase of 02- production.

Culture Media↗

PubMed1999

Mudskipper periophthalmodon schlosseri can repay oxygen debts in air but not in water

We determined aerial and aquatic oxygen uptake rates (&Mdot;o(2)) of Periophthalmodon schlosseri at rest and after exhaustive exercise at 30 degrees C. Resting &Mdot;o(2) in air (3.16 +/- 0.10 [SE] &mgr;mol/g/hr) was significantly higher than that in air saturated water (2.41 +/- 0.06 &mgr;mol/g/hr). When the fish was placed in air after 2 min exhaustive exercise, &Mdot;o(2) immediately increased about 2.5 times, thereby repaying oxygen debt of 5.0 &mgr;mol/g. In contrast, &Mdot;o(2) failed to show any significant elevation from the resting level as long as the fish was confined in water after exercise. However, when the fish was subsequently emerged, &Mdot;o(2) did increase above the resting level, amounting to an excess oxygen uptake of 5.5 &mgr;mol/g. These results demonstrated that the gas transport system of P. schlosseri is better adapted to air breathing and the mode of adaptation limited the capability for water breathing. J. Exp. Zool. 284:265-270, 1999. Copyright 1999 Wiley-Liss, Inc.

Journal Article↗

PubMed1999

Arterial blood gas levels and cardiovascular function during varying environmental conditions in a mudskipper, periophthalmodon schlosseri

Changes in blood gas levels, blood pressure and heart rate were studied in chronically cannulated mudskippers, Periophthalmodon schlosseri, subjected to air exposure (6 h), aquatic hypoxia with access to air (water PO2 <0.9 kPa, 6 h) and forced submersion in normoxic water (12 h) at 30 degrees C. Air exposure did not affect either blood O2 and had little effect on blood CO2 levels, but blood pH increased slightly, but significantly. Blood ammonia concentration was elevated sixfold during air exposure. Aquatic hypoxia caused no significant changes in blood gas levels. When the fish was forcibly submerged, blood O2 saturation decreased rapidly to approximately 30 %. Blood PCO2 and total CO2 also decreased, but blood pH was unaffected by forcible submersion. Air exposure did not affect blood pressure or heart rate. Aquatic hypoxia did not affect blood pressure but transiently increased heart rate. In contrast, forced submersion significantly depressed heart rate throughout the period of submersion, while blood pressure decreased only transiently. Upon emersion, the heart rate immediately increased to above the control level when the fish took its first air breath.

Journal Article↗

PubMed1998

Hemagglutinating activity of extracellular alkaline metalloendopeptidases from Vibrio sp. NUF-BPP1.

Alkaline metalloendopeptidase (metalloprotease) AP1 (48 kDa) from Vibrio sp. isolated from the intestine of a five-barred goatfish (Parupeneus trifasciatus) was reported in our previous paper to produce AP2 (36 kDa) by releasing a peptide fragment (molecular mass of about 12 kDa) from the C-terminal end of AP1 by autodigestion. AP1 strongly agglutinated fish (flounder, Paralichthys olivaceus) and rabbit erythrocytes, and weakly chicken erythrocytes. In contrast, AP2 had no significant hemagglutinating activity toward any erythrocytes tested, except for weak activity on flounder erythrocytes, suggesting that the C-terminal region of AP1 may be required for the strong hemagglutinating activity. The optimum temperature for the hemagglutinating activity of AP1 was found to be lower than that for the proteolytic activity. At acidic pHs (below pH 7.5), the hemagglutinating activity of AP1 decreased, and its pH profile resembled that of the proteolytic activity. The hemagglutinating activity of AP1 was not observed in the presence of o-phenanthroline or synthetic and proteinous substrates, but different kinds of saccharides and lipids had no effect. While the proteolytic activity of AP1 was not affected by CaCl2, the hemagglutinating activity of AP1 decreased with increases in CaCl2 concentrations. These results suggested that the hemagglutinating activity of these proteases (AP1 and AP2) was most likely caused by their proteolytic action on erythrocyte cell surfaces.

Animals↗

PubMed1997

Possible involvement of somatolactin in the regulation of plasma bicarbonate for the compensation of acidosis in rainbow trout

Somatolactin is a putative pituitary hormone of the growth hormone/prolactin family in fish. Its function is still unknown. The effects of environmental hypercapnia and hypoxia, acid (HCl) infusion and exhaustive exercise on plasma somatolactin levels were examined in the chronically cannulated rainbow trout to study the possible physiological roles of somatolactin. Respiratory acidosis induced by hypercapnia (2% CO2) did not affect plasma somatolactin level. In contrast, metabolic acidosis induced by acid infusion and exercise increased plasma somatolactin level. Blood pH was depressed to a similar extent by both types of acidosis, whereas plasma [HCO3-] was elevated by respiratory acidosis but reduced by metabolic acidosis. A moderate hypoxia (water PO2 9.3kPa) affected neither acidbase status nor plasma somatolactin level. A more severe hypoxia (water PO2 6.1kPa) resulted in metabolic acidosis accompanied by an apparent rise in plasma somatolactin level, although the difference in somatolactin level from the control value was not statistically significant. Somatolactin immunoneutralization retarded recovery of plasma [HCO3-] following acid infusion. These results indicate that somatolactin is involved in the retention of HCO3- during metabolic acidosis but not in the active accumulation of HCO3- for acidbase compensation of respiratory acidosis in rainbow trout Oncorhynchus mykiss.

Journal Article↗

PubMed1997

Generation of reactive oxygen species by raphidophycean phytoplankton.

Chattonella marina, a raphidophycean flagellate, is one of the most toxic red tide phytoplankton and causes severe damage to fish farming. Recent studies demonstrated that Chattonella sp. generates superoxide (O2-), hydrogen peroxide (H2O2), and hydroxyl radicals (.OH), which may be responsible for the toxicity of C. marina. In this study, we found the other raphidophycean flagellates such as Heterosigma akashiwo, Olisthodiscus luteus, and Fibrocapsa japonica also produce O2- and H2O2 under normal growth condition. Among the flagellate species tested, Chattonella has the highest rates of production of O2- and H2O2 as compared on the basis of cell number. This seems to be partly due to differences in their cell sizes, since Chattonella is larger than other flagellate species. The generation of O2- by these flagellate species was also confirmed by a chemiluminescence assay by using 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazin++ +-3-one (MCLA). All these raphidophycean flagellates inhibited the proliferation of a marine bacterium, Vibrio alginolyticus, in a flagellates/bacteria co-culture system, and their toxic effects were suppressed by the addition of superoxide dismutase (SOD) or catalase. Our results suggest that the generation of reactive oxygen species is a common feature of raphidophycean flagellates.

Animals↗

PubMed1996

Analysis of cardiac shunting in the turtle Trachemys (Pseudemys) scripta: application of the three outflow vessel model.

Blood distribution within the ventricle was analysed in acutely prepared turtles Trachemys scripta by measuring the oxygen concentration and flow rates of blood in the central vessels. Pulmonary (Qp) and systemic (Qs) blood flow rates were similar when total cardiac output (Qtot) was below 40 ml min-1 kg-1. Above this value, increments of Qtot were directed to the pulmonary circuit, with Qs levelling off at approximately 20 ml min-1 kg-1. When Qtot was larger than 40 ml min-1 kg-1, the systemic circuit was almost exclusively perfused by left atrial blood and systemic venous return was almost all directed towards the lungs. Blood oxygen levels and flow rates were consistently higher in the right aorta than in the left aorta. Blood movement within the ventricle, coupled with differences in ejection timing, is probably the decisive factor determining this pattern of blood distribution in the turtle heart.

Animals↗

PubMed1996

The mechanism of cardiac shunting in reptiles: a new synthesis.

The mechanism of cardiac shunting in reptiles is controversial. Recent evidence suggests that a right-to-left shunt in turtles results primarily from a washout mechanism. The mechanism that accounts for left-to-right (L-R) shunting is unresolved. This study used haemodynamic analysis and digital subtraction angiography to determine the mechanism of L-R cardiac shunting in the turtle Trachemys (Pseudemys) scripta. Animals were instrumented with ultrasonic blood flow probes (Transonic Systems, Inc.) for the measurement of total pulmonary blood flow and total systemic blood flow. In addition, catheters were inserted into the common pulmonary artery (PA), the systemic arteries, the left atrium and right atrium. These catheters were used for the measurement of blood pressure or for the infusion of radio-opaque material. Haemodynamic conditions were altered by electrical stimulation of the afferent (VAF) or efferent vagal nerves or by infusion of vasoactive drugs. Under control conditions, the peak systolic pressure in the systemic arteries was slightly higher than that in the PA (30.6 versus 28.3 mmHg; 4.08 versus 3.77 kPa), whereas diastolic pressure in the PA was significantly less than that in the systemic arteries (9.8 versus 24.4 mmHg; 1.31 versus 3.25 kPa). During VAF stimulation, the peak systolic pressures in the PA and aortae almost doubled. Diastolic pressure in the systemic arteries also doubled, but it increased by only 45% in the PA. Ejection of blood into the PA preceded that into the left aorta by 53 ms under control conditions. This difference increased (by as much as 200 ms) as the difference in the diastolic pressures between the two circulations increased during VAF stimulation. This resulted in the development of a large net L-R shunt. Under these conditions, digital subtraction angiography showed that the L-R shunt resulted from a combination of both washout and pressure mechanisms.

Angiography, Digital Subtraction↗

PubMed1992

Hydroxyl radical generation by red tide algae.

The unicellular marine phytoplankton Chattonella marina is known to have toxic effects against various living marine organisms, especially fishes. However, details of the mechanism of the toxicity of this plankton remain obscure. Here we demonstrate the generation of superoxide and hydroxyl radicals from a red tide unicellular organism, C. marina, by using ESR spectroscopy with the spin traps 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) and N-t-butyl-alpha-phenylnitrone (PBN), and by using the luminol-enhanced chemiluminescence response. The spin-trapping assay revealed productions of spin adduct of superoxide anion (O2-) (DMPO-OOH) and that of hydroxyl radical (.OH) (DMPO-OH) in the algal suspension, which was not observed in the ultrasonic-ruptured suspension. The addition of superoxide dismutase (500 U/ml) almost completely inhibited the formation of both DMPO-OOH and DMPO-OH, and carbon-centered radicals were generated with the disappearance of DMPO-OH after addition of 5% dimethyl sulfoxide (Me2SO) and 5% ethanol. Furthermore, the generation of methyl and methoxyl radicals, which are thought to be produced by the reaction of hydroxyl radical and Me2SO under aerobic condition, was identified using spin trapping with a combination of PBN and Me2SO. Luminol-enhanced chemiluminescence assay also supported the above observations. These results clearly indicate that C. marina generates and releases the superoxide radical followed by the production of hydroxyl radical to the surrounding environment. The velocity of superoxide generation by C. marina was about 100 times faster than that by mammalian phagocytes per cell basis. The generation of oxygen radical is suggested to be a pathogenic principle in the toxication of red tide to susceptible aquaculture fishes and may be directly correlated with the coastal pollution by red tide.

Cyclic N-Oxides↗

PubMed1989

Relationship between respiratory state and intracardiac shunts in turtles.

Injection of radioactively labeled microspheres and blood O2 analysis from central vascular sites was utilized to assess intracardiac shunts in the unanesthetized and unrestrained turtle, Pseudemys scripta, at 15 degrees C. Both methods indicated a simultaneously occurring right-to-left (R-L) and left-to-right (L-R) shunt during ventilation and apnea. During ventilation, the O2 method estimated a R-L shunt of 24%. In contrast, injection of microspheres during ventilation estimated a R-L shunt of 65%, which was significantly greater than the value determined from the O2 method. During apnea, both methods indicated a significantly larger R-L shunt. Estimates of the R-L shunt from O2 analysis were between 60 and 90% of the venous return. The R-L shunt estimated from the O2 content of left aortic arch blood was significantly greater than the value determined from right aortic arch blood. During apnea, the microsphere method estimated a R-L shunt of 79%, which was in the intermediate range calculated by the O2 method using two aortic arches. Our results verify previous reports of significant differences in R-L shunt levels between ventilation and apnea during the normal ventilatory cycle of turtles.

Animals↗

PubMed1988

Analysis of intracardiac shunting in the lizard, Varanus niloticus: a new model based on blood oxygen levels and microsphere distribution.

The central sites of the cardiovascular system (right and left aortic arches, RAo and LAo, pulmonary artery, PA, and right and left atria, RAt and LAt) were chronically and non-occlusively cannulated for an analysis of intracardiac shunting in Varanus niloticus. Oxygen partial pressure (PO2) and oxygen concentration (CO2) were significantly higher in right aortic blood than values determined in left aortic blood. The difference was larger in animals acclimated to 25 degrees C (RAo CO2 = 4.5 +/- 1.00 vol %, LAo CO2 = 3.8 +/- 1.14, X +/- SD, n = 19) than at 35 degrees C (RAo CO2 = 5.8 +/- 1.24, LAo CO2 = 5.4 +/- 1.35, n = 18) (P less than 0.001 for both temperatures, paired t-test). These data are explained by a new model describing the differential shunting patterns of the two aortae in addition to the conventional overall right-to-left and left-to-right shunt fractions. This model was solved on the basis of blood gas data collected by simultaneous multiple-site gas analysis, together with data on the differential blood flow in the central vascular system, collected by application of the microsphere method. At 35 degrees C both right-to-left and left-to-right shunts were relatively small (about 9%), with the right-to-left shunt fraction directed exclusively into the left aorta. Thus right aortic blood represented left atrial blood, whereas left aortic blood was composed of 80% left atrial and 20% right atrial blood. Ninety percent of the pulmonary arterial blood was derived from the right atrium and 10% from the left atrium. At 25 degrees C the composition pattern of effluent blood for each vessel was similar, the absolute flow distribution, however, was different from that at 35 degrees C. These findings are discussed with respect to their significance and compatibility with the wash-out shunt model.

Animals↗

PubMed1988

In vivo analysis of partitioning of cardiac output between systemic and central venous sinus circuits in rainbow trout: a new approach using chronic cannulation of the branchial vein.

Freshwater-acclimated rainbow trout were chronically and non-occlusively cannulated in the dorsal aorta (DA), sinus venosus (SV) and branchial vein (BV), the latter returning the blood perfusing the central venous sinus (CVS) of the gill after being shunted away from the systemic circuit. After recovery, blood samples from these three sites were analysed for haematocrit (Hct) and [Hb]. Branchial venous blood was found to have considerably lower Hct and [Hb] (Hct = 3.5 +/- 3.1%; [Hb] = 1.04 +/- 0.75 g 100 ml-1) than systemic blood (DA: Hct = 24.3 +/- 8.9%, [Hb] = 6.54 +/- 2.90 g 100 ml-1; SV: Hct = 23.1 +/- 8.8%, [Hb] = 6.15 +/- 2.55 g 100 ml-1; means +/- S.D. N = 8), which strongly suggests that plasma skimming occurred at arteriovenous anastomoses and arterioles draining into the CVS. The partitioning of cardiac output, calculated on the basis of the [Hb] data, showed that the systemic flow accounted for 93 +/- 4.6% (N = 7) of the total cardiac output with only 7 +/- 4.7% of cardiac output being diverted into the CVS, thus bypassing the systemic vasculature. Previous results using in vitro experiments are compared with the present data in an evaluation of the usefulness of the isolated perfused gill and head preparations in the experimental analysis of circulatory physiology in fish gills.

Animals↗

PubMed1986

Autonomic nervous control of the circulatory system in the air-breathing fish Channa argus.

The control of the cardiovascular system with particular emphasis on the regulation of blood distribution in the gills and air-breathing organ was studied in the air-breathing teleost Channa argus. Perfused head preparations were used in addition to experiments with isolated strip preparations of arteries and heart chambers. The distribution of adrenergic nerves was investigated using Falck-Hillarp fluorescence histochemistry. This preliminary study shows an adrenergic control system composed of chromaffin cells and adrenergic nerves similar to that found in other teleosts investigated, although the systemic arteries (coeliac artery, dorsal aorta and the vasculature of the air-breathing organ) appear to lack an adrenergic innervation. The reactions of isolated artery strip preparations to acetylcholine and adrenaline resemble those seen in other teleosts, and there is a prominent inhibitory effect of L-isoprenaline suggestive of arterial beta-adrenoceptors. The general vascular resistance of the gill apparatus-air-breathing organ increases in response to acetylcholine or adrenaline, and there is a redistribution of perfusion flow from the air-breathing organ circuit (anterior venous outflow from the first and second pair of gills and the air-breathing organ) to the general systemic circuit (dorsal aortic outflow from the third and fourth pair of gills). Stimulation of the vagal branch entering the air-breathing organ mimics the effects of acetylcholine or adrenaline. This innervation is probably non-adrenergic since no adrenergic nerve fibres could be demonstrated in the vasculature of the air-breathing organ using the histochemical technique. An adrenergic control of the vasculature of the air-breathing organ is not likely, since the concentration of adrenaline needed to affect the vasculature is not reached in the plasma even during "stress".

Acetylcholine↗

PubMed1983

Blood oxygen levels and acid-base status following air exposure in an air-breathing fish, Channa argus: the role of air ventilation.

1. Acute air exposure of an air-breathing fish Channa argus immediately induced hypercapnic acidemia while total CO2 content of blood remained unchanged. Upon reimmersion, paco2 and pHa quickly restored to pre-exposure levels followed by gradual rise of [HCO-3]. 2. Artificial air ventilation of air-exposed fish restored acid-base status and greatly depressed voluntary air ventilatory movements. We conclude that the major cause of acid-base disturbances occurring during air exposure is the reduced air convection.

Acid-Base Equilibrium↗