The effect of diphenylhydantoin on LD 50 of ouabain was investigated in frogs, using "one hour frog method". LD50 of ouabain in control group was 1.90 microg/10g. A dose of 100 microg/10g diphenylhydantion did not affect the systemic manifestations of the frogs, but increase the LD50 of ouabain to 2.60 microg/10g. The difference of LD50 of ouabain and potency ratio between control group and diphenylhydantoin-treated group was statistically significant.
Lethal Dose 50 ; Ouabain* ; Phenytoin*

ID clarify the effects of ouabain on the ERG c-wave, isolated chick retinas were exposed to different concentrations of ouabain and the results noted. Although the c-wave was abolished at a highe. dose of ouabain (10(-4)M), its amp1itude increased in the presence of ouabain at a concentration of 10(-7)M, which was within the range of clinical use of the cardiac glycoside. On the other hand, the standing potential of the retina did not change appreciably until 10(-6)M and then decreased gradually at higher concentrations.In the presence of 10(-4)M ouabain, the concentration which completely blorked Na-K-ATPase, both the c-wave and the standing potential were almost abolished. These phenomena were more conspicuous when ouabain was applied to the vitreous side rather than the choroidal side. In the presence of 10(-7)M ouabain, the light sensitivity of the retina was elevated to 0.5 log unit and the maximum response increased about 30%. This may be a sign of visual complications of ouabain, such as metachromatopsia.
Animals ; Chickens ; Electroretinography ; Ouabain/*pharmacology ; Photic Stimulation ; Retina/*drug effects

It has been reported that ascorbic acid[AA]appears to be actively taken up by the corneal endothelium and protect the endothelium against harmful effects of the oxidative reactions. To investigate the effect of ascorbic acidon the corneal endothelial function, rabbit`s corneas were mounted in the in vitro specular microscope. Corneal endothelium was perfused with ascorbic acid, then switched to AA plus ouabain solution, and vice versa. Also, phloretin was perfused onto the endothelium with AA and ouabain. Andcorneal endothelium was perfused with GBR or AA solution followed by perfusion with ouabain. Corneal thickness was measured during the perfusion and the corneal swelling rate calculated. Corneal endothelial permeability was also measured after perfusion of ascorbic acid. Perfusion with AA showed no corneal swelling, but swelling rate was even lower than GBR control. Corneal endothelial permeability did not change upon AA perfusion. In corneas preperfused with ouabain, AA added to ouabain solution decreased corneal swelling rates induced by ouabain solution[19.9 vs. 40.5 micrometer/hr]. The corneas preperfused with AA also showed decreased swelling rates with subsequent perfusion of ouabain added to AA solution[21.7 vs.28.6 micrometer/ hr]. Phloretin inhibited the effect of AA.However, when ouabain was removed, the corneal swelling plateaued but did not return to baseline thickness in both AA and GBR perfusion.The results of this study showed that AA can increase corneal endothelial pump function and reduce corneal swelling caused by ouabain.
Ascorbic Acid* ; Cornea ; Endothelium ; Endothelium, Corneal ; Ouabain ; Perfusion ; Permeability ; Phloretin

The effect of Carteolol hydrocholride on the Na-K ATPase of anterior capsule and epithelium of cattle lens has been investigated. The experiments were also designed to determine the action of Carteolol hydrochloride on the Na-K ATPase activity in anterior capsule and epithelium of cattle lens. The following results were observed. 1. The predominent location of Na-K ATPase was located in the epithelim and NaK ATPase was closely related with active trasport system of sodium and potassium in the lens epithelium. 2. The activity of Na-K ATPase of cattle lens epithelium was alomst totally inhibited by Ouabain. 3. Beta-adrenergic blocking agent(Carteolol hydrochloride) in the epithelium of cattle lens was shown to be actively transported by the ATPase. 4. The Na-K ATPase activity of cattle lens epithelium was inhibited by Carteolol hydrochloride.
Adenosine Triphosphatases* ; Animals ; Biological Transport, Active ; Carteolol* ; Cattle* ; Epithelial Cells* ; Epithelium ; Membranes* ; Ouabain ; Potassium ; Sodium

BACKGROUND AND OBJECTIVES: Controversy exists regarding the role of endogenous ouabain in the pathogenesis of DOCA-salt induced hypertension. The purpose of this study was to investigate the role of endogenous ouabain in the development of hypertension in DOCA-salt rats. MATERIALS AND METHODS: The mean blood pressure and heart rate were recorded in 1, 2 and 4 week old control and DOCA-salt treated rats. The endogenous levels of ouabain in the plasma, hypothalamus, pituitary and adrenal glands of the 1, 2 and 4 week old control and DOCA-salt treated rats were also measured using a radioimmunoassay. RESULTS: The mean blood pressures in the 2 and 4 week old DOCA-salt treated rats were significantly higher than those of the controls. There was no significant change in the heart rate between the DOCA-salt treated and control groups. In the 4 week old DOCA-salt treated rats, the endogenous level of ouabain in the adrenal glands was higher than that in the control rats, but this was only weakly significant. The endogenous level of ouabain in the hypothalamus was significantly higher in the 1 week old DOCA-salt treated rats than in the control, but this significance disappeared in the 2 and 4 week old DOCA-salt treated rats. CONCLUSION: These results suggest that the endogenous level of ouabain contributes to the development and maintenance of high blood pressure in DOCA-salt rats. Further studies will be required to elucidate the relationship between the endogenous level of ouabain and DOCA-salt hypertension.
Adrenal Glands ; Animals ; Blood Pressure ; Heart Rate ; Hypertension ; Hypothalamus ; Ouabain* ; Plasma ; Radioimmunoassay ; Rats*

OBJECTIVE: The oligosaccharide moieties of glycoproteins and proteoglycans have a vital function in blastocyst differentiation. Concanavalin (ConA), a lectin, is known to bind on the preimplantation embryos, especially on blastocyst. In this study, we investigated whether ConA can modulate the trophoblast development and about the regulating mediator. Also, we investigated whether expansion is enough for hatching procession of the mouse blastocyst. METHOD: Embryos were collected at 72 h post hCG injection and chemicals were treated after 24 h (96 hr post hCG injection). ConA or calcium ionophore A23187 were exposed to blastocyst and than analysis the developmental process for 48 hr. Intracellular free-Ca2+ concentration in trophectoderm was measured with confocal laser microscope after exposing to ConA or calcium ionophore A23187. ConA-pretreated blastocyst exposed to the calcium ionophore A23187 and then analyzed the developmental process. Otherwise ouabain was treated to the blastocyst to block the Na+/K+-ATPase activity. RESULTS: In contrast to the control blastocyst, the ConA-exposed blastocysts developed beyond the expansion stage with significantly high rate (90.4%) at 12 h post administration. ConA induced an increase the intracellular Ca2+ concentration in trophectoderm. Calcium ionophore A23187 also stimulated expansion of blastocyst. Most of the control blastocysts developed to the hatching stage at 144 h post hCG injection. However, strongly 65% of the ConA-exposed embryos were arrested at expanded stage at same time point. The developmental progression rates to hatching stage of both ConA- and calcium ionophore A23187-expose blastocysts were significantly lower than that of the control. However ConA-pretreated embryos developed to the hatching stage like control embryos. Ouabain showed a tendency to delayed the progress to expansion stage but did not inhibit the development to the hatching stage. CONCLUSION: ConA-mediated expansion is the result of the increase of intracellular free-calcium in blastocyst stage embryo. It is suspected that expansion of the blasocyst is a essential indirect factor in hatching and the calcium may triggering the cellular mechanisms for the both expansion and hatching progression.
Animals ; Blastocyst* ; Calcimycin ; Calcium* ; Concanavalin A* ; Embryonic Structures ; Glycoproteins ; Mice* ; Ouabain ; Proteoglycans ; Trophoblasts

Intracellular ions which have a major role in cellular function have been reported to affect repair of radiation damage. Recently it has been reported that ouabain sensitizes A549 tumor cells hut not CCL-120 normal cells to radiation. Ouabain inhibits the Na+-K+-pump rapidly thus it increases intracellular Na concentration. Vanadate which is distributed extensively in almost all living organisms in known to be a Na+-K+-ATPase inhibitors. This study was performed to see any change in radiosensitivity of tumor cell by vanadate and any role of Na+-K+-ATPase in radiosensitization. Experiments have been carried out by pretreatment with vanadate in human cell line(A549, JMG) and mouse cell line(L1210, spleen). For the cell survival MTT assay was performed for A549 and JMG cell and trypan blue dye exclusion test for L120, and spleen cells. Measurements of Na+-K+-ATPase activity in control, vanadate treated cell, radiation treated cell (9 Gy for A549 and JMG, 2 Gy for L1201, spleen), and combined 10-6 M vanadate and radiation treated cells were done. The results were summarized as follows. 1. L1210 cell was most radiosensitive, and spleen cell and JMG cell were intermediate, and A549 cell was least radiosensitive. 2. Minimum or cytotoxicity was seen with vanadate below concentration of 10-6 M. 3. In A549 cells there was a little change in radiosensitivity with treatment of vanadate. However radiation sensitization was shown in low dose level of radiation i. E. 2-Gy. In JMG cells no change in radiosensitivity was noted. Both L1210 and spleen cell had radiosensitization but change was greater in tumor cell. 4. Na+-K+-ATPase activity was inhibited significantly in tumor cell by treatment of vanadate. 5. Radiation itself inhibited Na+-K+-ATPase activity of tumor cell with high Na+- K+-ATPase concention. Increase in radiosensitivity by vanadate was closely associated with original Na+-K+-ATPase contents. From the above results vanadate had little cytotoxicity and it sensitized tumor cells to radiation. Inhibitory effect of vanadate on Na+-K+-ATPase activity might be one of the contributing factors for radiosensitization to tumor cells which has greater enzyme activity than that of normal cell. It was suggested vanadate could be used as a potential radiosensitizer for tumor cells.
Animals ; Cell Survival ; Humans ; Ions ; Mice ; Ouabain ; Radiation Tolerance* ; Spleen ; Trypan Blue ; Vanadates*

The effect of saponin on the Na+ - K+ ATPase of anterior capsule and epithelium of cattle lens has been investigated. The experiment also designed to determine the cation of saponin on the Na+ - K+ ATPase activity in anterior capsule and epithelium of cattle lens. The following results are observed. 1. The Na+ - K+ ATPase activity of cattle lens epithelial cell membrane is inhibited by low concentration of saponin, but increased by high concentration. The strongest activity showed at the dose of 50mg% saponin concentration. 2. The activating effect of saponin on the cattle epithelial cell membrane Na+ - K+ ATPase activity is inhibited by ouabain, but the stimulation of the Mg++ ATPase by high concentration of saponin is not inhibited by ouabain. 3. The activity ratio of Na+ - K+ ATPase by high concentration of saponin is increased by raising the sodium concentration but decreased by raising the potassium concentration. 4. The Na+ - K+ ATPase activity is increased by small amount of calcium but completely inhibited by over 5mM concentration of calcium.
Adenosine Triphosphatases* ; Animals ; Calcium ; Cataract ; Cattle* ; Epithelial Cells* ; Epithelium ; Membranes ; Ouabain ; Potassium ; Saponins* ; Sodium

The author had investigated the effects of Na-K-ATPase activity on the incubation period, ATP and enzyme concentration, and various electrolytes in brain homogenate of Korean native goat. The results were summarized as follows: 1) The initial velocity of Na-K-ATPase reaction was in proportion to incubation period and enzyme concentration. However, the optimal reaction was found at 2 mM ATP, 50-100 mM Na, 5-10 mM K, 2-4 mM Mg, and pH 7.4 in medium. 2) Na concentration for the optimal activity of Na-K-ATPase was varied by K concentration and the ratio of K to Na concentration for optimal activity was roughly 1 to 5. 3) The Na-K-ATPase activity was inhibited by Ca. The inhibitory effect by Ca was an apparently mixed type with respect to Na activation of the Na-K-ATPase, by reducing apparent Vmax but increasing the apparent Km for Na. 4) The Na-K-ATPase activity was inhibited by cadmium and ouabain, The Km for cadmium and ouabain was 8+/-10 M and 1+/-10 M, respectively. The general characteristics of Na-K-ATPase activity in goat cerebral cortex were similar to that of another animals.
Adenosine Triphosphate ; Animals ; Brain ; Cadmium ; Cerebral Cortex* ; Electrolytes ; Goats* ; Hydrogen-Ion Concentration ; Ouabain

Effect of Ouabain administration on aqueous humor dynamics were studied in albino rabbits. Intravenous injection of 0.1 mg/kg of ouabain in 10 rabbits produced significant reduction of intraocular pressure. The highest lowering of intraocular pressure occurred 70 minutes after the injection, its magnitude being 35.54% of the original pressure. The intraocular pressure returned to its original pressure 117.0 minutes after the injection. Perilimbal suction cup studies revealed 26.92 % reduction in aqueous flow. Intravitreal injection of O.1 micro gram of ouabain dissolved in 0.02 ml performed in 5 rabbits' eyes produced marked lowering of intraocular pressure. The maximum tension lowering effect (60.15%) was observed 4.6 days after the administration, and the tension recovered 15.8 days after the injection. Perilimbal suction cup studies showed 29.48 % reduction in aqueous flow. It is concluded that Na-K ATPase play an important role in the aqueous formation and that ouabain lowers the intraocular pressure by supressing this enzyme. Intravitreal administration has more powerful tension-lowering effect than intravenous injection.
Adenosine Triphosphatases ; Aqueous Humor ; Injections, Intravenous ; Intraocular Pressure* ; Intravitreal Injections ; Ouabain* ; Rabbits* ; Suction