We have investigated the two types of voltage-dependent outward potassium (K) currents, i.e. delayed rectifier K current (I-K(V)) and 'A-like' transient outward K current (I-to) with patch-clamp technique in single smooth muscle cells (SMCs) isolated from rabbit basilar artery, and investigated the characteristics of them. The time-courses of activation were well fitted by exponential function raised to second power (n-2) in I-K(v) and fourth power (n-4) in I-to. The activation, inactivation and recovery time courses of I-to were much faster than that of I-K(V). The steady-state activation and inactivation of I-K(V) was at the more hyperpolarized range than that of I-to contrary to the reports in other vascular SMCs. Tetraethylammonium chloride (TEA; 10 mM) markedly inhibited I-K(V) but little affected 1-to. 4-Aminopyridine (4-AP) had similar inhibitory potency on both currents. While a low concentration of Cd-2+ (0.5 mM) shifted the current-voltage relationship of I-to to the positive direction without change of maximum conductance, Cd-2+ did not cause any appreciable change for I-K(V).
4-Aminopyridine ; Basilar Artery* ; Muscle, Smooth* ; Myocytes, Smooth Muscle* ; Patch-Clamp Techniques ; Potassium* ; Tetraethylammonium

The authors have recently experienced 3 cases of ocular toxoplasmosis. The diagnosis was based on typical ocular lesions and hemagglutination test for toxoplasmosis. In addition to some clinical observations, a brief review of literature has been described.
Diagnosis ; Hemagglutination Tests ; Toxoplasmosis ; Toxoplasmosis, Ocular*

By using differential display, we identified one of the genes encoding the multi-subunit complex protein V-ATPase, c subunit gene (ATP6L), and showed alterations of the gene expression by oxidative stresses. Expression of the ATP6L gene in Neuro-2A cells was increased by the treatment with H2O2 and incubation in hypoxic chamber, implying that the expression of the ATP6L gene is regulated by oxidative stresses. To examine mechanisms involved in the regulation of the gene expression by oxidative stresses, the transcriptional activity of the rat ATP6L promoter was studied. Transcription initiation site was determined by primer extension analysis and DNA sequencing, and promoter of the rat ATP6L and its deletion clones were constructed in reporter assay vector. Significant changes of the promoter activities in Neuro-2A cells were observed in two regions within the proximal 1 kbp promoter, and one containing a suppressor was in -195 to -220, which contains GC box that is activated by binding of Sp1 protein. The suppression of promoter activity was lost in mutants of the GC box. We confirmed by electrophoretic mobility shift and supershift assays that Sp1 protein specifically binds to the GC box. The promoter activity was not changed by the H2O2 treatment and incubation in hypoxic chamber, however, H2O2 increased the stability of ATP6L mRNA. These data suggest that the expression of the ATP6L gene by oxidative stresses is regulated at posttranscriptional level, whereas the GC box is important in basal activities of the promoter.
Animals ; Clone Cells ; Gene Expression* ; Hydrogen Peroxide ; Oxidative Stress* ; Rats ; RNA, Messenger ; Sequence Analysis, DNA ; Transcription Initiation Site ; Vacuolar Proton-Translocating ATPases*

OBJECTIVES: To understand the importance of the serum level of Ca-125 among pelvic mass, we performed a study. METHOD: From January to December 1998, we performed the study. Before hysterectomy, we performed a blood sampling to know the serum level of Ca-125, After hysterectomy, we weighed the uterus and measured the thickness of endometrium and other histologic characteristics. RESULTS: We performed my research to 80 peoples. The relation between uterine weight and the serum level of Ca-125 is little, if ever(R2=0.0007), and the relation between the thickness of endometrium and the serum level of Ca-125 is also little, if ever(R2=0.0353). The relation between leiomyoma, the cycle of endometrium and the serum level of Ca-125 were also little, but there was a close relationship between adenomyosis and the serum level of Ca-125. CONCLUSION: There was little relationship between uterine weight and the serum level of Ca-125.
Adenomyosis ; Endometrium ; Female ; Humans ; Hysterectomy* ; Leiomyoma ; Uterus

delta12-Prostaglandin J2 (delta12-PGJ2) is one of cyclopentenone prostaglandins. The delta12-PGJ2 is known to induce apoptosis of tumor cells, however, it's action mechanism is not clear. It has recently been reported that STAT3 is involved in tumorigenesis. In the present study, we investigated the role of STAT3-interacting protein (STIP1) in the cytotoxicity of delta12-PGJ2, since STIP1 was recently reported as a modulator of STAT3 activation by specifically binding to inactive (unphosphorylated) STAT3. The effect of delta12-PGJ2 was observed in stably overexpressing Neuro-2A cells transfected with full cDNA of STIP1, and cytotoxicity of delta12-PGJ2 in the transfected cells was increased, compared with the vector control cells. The cytotoxicity of delta12-PGJ2 treatment was significantly accentuated by pretreatment of the STIP1-transfected cells with protein kinase inhibitor, genistein, and less activation of STAT3 in STIP1-transfected cells was shown, compared with the vector control cells. Expression of bax was also increased in the STIP1-transfected cells. These data suggest that STIP1 inhibits cell growth via inhibition of STAT3 activation in delta12-PGJ2 treatment.
Apoptosis ; Carcinogenesis ; Cell Death* ; DNA, Complementary ; Genistein ; Prostaglandins ; Protein Kinases

This study was designed to identify and characterize Na+ -activated K+ current (I(K(Na))) in guinea pig gastric myocytes under whole-cell patch clamp. After whole-cell configuration was established under 110 mM intracellular Na+ concentration ([Na+]i) at holding potential of -60 mV, a large inward current was produced by external 60 mM K+([K+] degree). This inward current was not affected by removal of external Ca2+. K+ channel blockers had little effects on the current (p>0.05). Only TEA (5 mM) inhibited steady-state current to 68+/-2.7% of the control (p<0.05). In the presence of K+ channel blocker cocktail (mixture of Ba2+, glibenclamide, 4-AP, apamin, quinidine and TEA), a large inward current was activated. However, the amplitude of the steadystate current produced under [K+]degree (140 mM) was significantly smaller when Na+ in pipette solution was replaced with K+ - and Li+ in the presence of K+ channel blocker cocktail than under 110 mM [Na+]i. In the presence of K+ channel blocker cocktail under low Cl- pipette solution, this current was still activated and seemed K+ -selective, since reversal potentials (E(rev)) of various concentrations of [K+]degree-induced current in current/voltage (I/V) relationship were nearly identical to expected values. R-56865 (10-20 microgram), a blocker of IK(Na), completely and reversibly inhibited this current. The characteristics of the current coincide with those of IK(Na) of other cells. Our results indicate the presence of IK(Na) in guinea pig gastric myocytes.
Tetraethylammonium Compounds/pharmacology ; Stomach/*physiology ; Sodium/metabolism/*pharmacology ; Potassium Channels/*physiology ; Potassium Channel Blockers/pharmacology ; Myocytes, Smooth Muscle/*physiology ; Membrane Potentials ; Male ; Guinea Pigs ; Female ; Chlorides/pharmacology ; Calcium/metabolism ; Animals

We explore the question of whether adenosine 5'-triphosphate (ATP) acts as an excitatory neurotransmitter in guinea-pig gastric smooth muscle. In an organ bath system, isometric force of the circular smooth muscle of guinea-pig gastric antrum was measured in the presence of atropine and guanethidine. Under electrical field stimulation (EFS) at high frequencies (>20 Hz), NO-mediated relaxation during EFS was followed by a strong contraction after the cessation of EFS (a "rebound-contraction"). Exogenous ATP mimicked the rebound-contraction. A known P2Y-purinoceptor antagonist, reactive blue 2 (RB-2), blocked the rebound-contraction while selective desensitization of P2x-purinoceptor with alpha, beta-MeATP did not affect it. ATP and 2-MeSATP induced smooth muscle contraction, which was effectively blocked by RB-2 and suramin, a nonselective P2-purinoceptor antagonist. Particularly, in the presence of RB-2, exogenous ATP and 2-MeSATP inhibited spontaneous phasic contractions, suggestingthe existence of different populations of purinoceptors. Both the rebound-contraction and the agonist-induced contraction were not inhibited by indomethacin. The rank orders of agonists' potency were 2-MeSATP > ATP gtoreq UTP for contraction and alpha, beta-MeATP gtoreq beta, gamma-MeATP for inhibition of the phasic contraction, that accord with the commonly accepted rank order of the classical P2Y-purinoceptor subtypes. Electrical activities of smooth muscles were only slightly influenced by ATP and 2-MeSATP, whereas alpha, beta-MeATP attenuated slow waves with membrane hyperpolarization. From the above results, it is suggested that ATP acts as an excitatory neurotransmitter, which mediates the rebound-contraction via P2Y-purinoceptor in guinea-pig gastric antrum.
Adenosine Triphosphate* ; Adenosine* ; Atropine ; Baths ; Guanethidine ; Indomethacin ; Membranes ; Muscle, Smooth ; Neurotransmitter Agents* ; Pyloric Antrum* ; Receptors, Purinergic ; Relaxation ; Stomach ; Suramin ; Uridine Triphosphate

The spontaneous contractions of gastric smooth muscles are regulated by slow waves, which are modulated by both nervous system and humoral agents. This study was designed to examine the effects of Prostaglandin E2 (PGE2) on the contractile and electrical activities of antral smooth muscles in guinea-pig stomach, using an intracellular recording technique. To elucidate the underlying mechanism for its effect on contractility, ionic currents were also measured using a whole-cell patch clamp method. The basal tone by PGE2 was variable, whereas the magnitude of phasic contractions was reduced (19.0 +/- 2.1%, n=19). The resting membrane potentials were hyperpolarized (-4.4+/-0.5 mV, n=10), and plateau potentials were lowered (-2.9+/-0.5 mV, n=10). In most cases, however, the initial peak potentials of slow waves were depolarized more by PGE2 than those of control. The frequency of the slows wave was increased from 5.7+/-0.2 cycles/min to 6.5+/-0.2 (n-22). Voltage-operated Ca2+ currents were decreased by PGE2 (n=5). Voltage-operated K+ currents, both Ca-dependent and Ca-independent, were increased (n-5). These results suggest that PGE2 plays an important role in the modulation of gastric smooth muscle activities, and its inhibitory effects on the contractility and activities of slow waves are resulted from both decrease of Ca2+ currents and increase of K+ currents.
Dinoprostone* ; Membrane Potentials ; Muscle, Smooth ; Nervous System ; Stomach*

OBJECTIVES: Several reports demonstrated that ethanol administration impairs the DNA synthesis in rat hepatocytes. Also, it has been demonstrated that prostaglandin (PG) helps prevent membrane damage by hepatotoxic chemicals. In this study, the authors examined PG's effects on the toxicity of ethanol in the primary culture of rat regenerations. METHODS: We examined two kinds of parameters, i.e., DNA synthesis and lipid peroxidation in the primary culture of rat hepatocytes. Hepatocytes were isolated by the collagenase perfusion method. The rate of DNA synthesis was determined by pulse-labelling cultured cells with [3H]-thymidine. Incorporation of (3H)-thymidine was determined by liquid scintillation spectrophotometer. DNA content was measured by the fluorescence spectrophotometer. The lipid peroxidation was assayed with spectrophotometer. RESULTS: The results were as follows: 1) PG family (PGA1, PGD2, PGE1, PGE2, PGG2a, PGI2 & Thromboxane B2) stimulated the DNA synthesis of hepatocytes (especially PGD2 and PGE1), 2) ethanol decreased DNA synthesis by clear dose-dependent manner, 3) the combined treatment of PGD2 or PGE1, prevents the decreasing of DNA synthesis, which was induced by ethanol, 4) in ethanol treatment, lipid peroxidation was decreased significantly, but PGD2, PGE1 and PGA1 were not affected, and 5) PGD2, PGE1 and PGA1 decreased lipid peroxidation with ethanol, significantly. CONCLUSIONS: From these results, we concluded that PG could be useful for the treatment of degenerative liver disease and alcohol-induced liver disease in the assumption that further studies on the action mechanisms of PG will continue.
Animal ; Cells, Cultured ; DNA/biosynthesis ; Drug Interactions ; Ethanol/toxicity* ; Lipid Peroxidation/drug effects ; Liver/metabolism ; Liver/drug effects* ; Prostaglandins, Synthetic/pharmacology* ; Rats

BACKGROUND AND OBJECTIVES: The restrictive filling pattern (RFP) is accepted as a poor prognostic marker in congestive heart failure (CHF) patients. But, recently the RFP has been categorized into various prognosis subgroups by their clinical signs or echocardiographic markers with loading manipulation. But, in critically ill or severe dyspneic patients these loading manipulations are not practical to apply. Therefore, we tried to establish simple, reliable prognostic echocardiographic variables in CHF patients with the RFP. MATERIALS AND METHOD: 40 patients with the RFP were observed for 35+/-19 months after echocardiographic examination. We obtained baseline peak early (E), late (A) mitral inflow velocities, E/A ratio, deceleration time of E velocity (DT), peak early (E'), late (A') diastolic mitral annulus velocities, E'/A' ratio, reversibility of the RFP. The reversibility of RFP was defined as E/A ratio reverse (<1) during the Valsalva's maneuver. With the clinical and survival data during follow up period, we established significant prognostic variables in these patients. RESULTS: In univariate analysis, low systolic blood pressure (p=0.013), low A velocity (p=0.044), low A' velocity (p=0.028) and the irreversibility of RFP (p=0.024) were significant drastic prognosis variables. Especially, patients with A velocity <0.32 m/sec or A' velocity <0.04 m/sec showed significantly higher mortality. CONCLUSION: In CHF patients with the RFP, A velocity and A' velocity are very practical prognostic echocardiographic variables and patients with the low A velocity (<0.32 m/sec) or the low A' velocity (<0.04 m/sec) showed higher mortality rate.
Blood Pressure ; Critical Illness ; Deceleration ; Echocardiography ; Follow-Up Studies ; Heart Failure* ; Heart* ; Humans ; Mortality ; Prognosis* ; Valsalva Maneuver