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Oncogenic types of human papilloma virus (HPV) are known to be closely associated with cervical carcinoma. On the other hand, the oncogenic process is associated with various abnormalities in the mechanisms of cellular regulation. In this study, we detected the expressions of p53 and p21 proteins in cervical lesions by immunohistochemical techniques, and examined the relationship with HPV infection as well as the clinical usefulness of the results. Cervical biopsy specimens from 107 cases of cervical lesions were studied. HPV-DNA was detected by the hybrid capture method using probe A for low oncogenic types and probe B for high oncogenic types. Anti p21, anti-p53 antibodies were used to perform immunostaining. Point mutation in the p53 gene was analyzed by the DGGE method. High oncogenic HPV types were detected at high frequencies in CIN and SCC. In lesions associated with high oncogenic HPV, p53 protein was detected in 33.4% of the lesions and p21 protein in 36.3%. The p53 gene was analyzed in all cases, and point mutation was not detected. No relation was detected between HPV infection and p53/p21 protein expression. Since mutation was not found in the p53 gene, the p53 protein expressed was considered to be wild-type, which is suspected to play a role in inhibiting disease progression in some cases.

Transcription factors p53 and nuclear factor kappaB (NF-kappaB) have been implicated in apoptosis induced by DNA-damaging agents, but the relationship between these two factors at the molecular level is largely unknown. We have isolated apoptosis-resistant mutant sublines from a radiosensitive mouse lymphoma 3SB cell line that undergoes p53-depen-dent apoptosis after X-ray irradiation, and we have analyzed the NF-kappaB activity. Two of these apoptosis-resistant sublines expressed mutant p53 protein and exhibited a defect in the induction of cyclin-dependent kinase inhibitor p21 after X-ray irradiation. A decrease in the DNA binding activity of NF-kappaB was observed in the parental 3SB cells after exposure to X-rays, whereas the same activity was unaffected by radiation in the two mutant sublines. A similar down-regulation of NF-kappaB activity by X-rays was observed in thymocytes derived from p53 wild-type and heterozygous mice, but not in thymocytes from p53 homozygous knock-out mice. These results suggest that NF-kappaB inactivation is p53 dependent and is required for X-ray-induced apoptosis in thymic lymphoma cells and normal thymocytes.

The heterogeneity of the liver parenchyma in relation to uric acid production from adenosine was investigated using the bivascularly perfused rat liver in the anterograde and retrograde modes. Adenosine was infused in livers from fed rats during 20 min at four different concentrations (20, 50, 100 and 200 microM) according to four experimental protocols as follows: (A) anterograde perfusion, with adenosine infusion into the portal vein; (B) anterograde perfusion, with adenosine in the hepatic artery, (C) retrograde perfusion, with adenosine in the hepatic vein; (D) retrograde perfusion, with adenosine in the hepatic artery. With protocols A, B, and D uric acid production from adenosine was always characterized by initial bursts followed by progressive decreases toward smaller steady-states. With protocol C the initial burst was present only when 200 microM adenosine was infused. The initial bursts in uric acid production were accompanied by simultaneous increases in the ratio of uric acid production/adenosine uptake rate. These initial bursts are thus representing increments in the production of uric acid that are not corresponded by similar increments in the metabolic uptake rates of adenosine. Global analysis of uric acid production revealed that the final steady-state rates were approximately equal for all infusion rates with protocols A, B and C, but smaller with protocol D. This difference, however, can be explained in terms of the differences in accessible cellular spaces, which are much smaller when protocol D is employed. When the analysis was performed in terms of the extra amounts of uric acid produced during the infusion of adenosine, where the initial bursts are also taken into account, different dose-response curves were found for each experimental protocol. These differences cannot be explained in terms of the accessible cell spaces and they are likely to reflect regional heterogeneities. From the various dose-response curves and from the known characteristics of the microcirculation of the rat liver it can be concluded that the initial bursts in uric acid production are generated in periportal hepatocytes. The reason for this heterogeneity could be related to the metabolic effects of adenosine, especially to oxygen uptake inhibition, which is likely to produce changes in the ATP/AMP ratios.

Conflicting results have been reported concerning the effect of endoscopic injection sclerotherapy upon the long-term survival of cirrhotic patients with esophageal varix. The recurrence and rebleeding of esophageal varices seems to be an important factor influencing long-term survival. We investigated the long-term survival of patients after complete eradication of esophageal varices.Forty patients treated by endoscopic injection sclerotherapy for acute esophageal variceal bleeding were studied. The recurrence rate of varices and the long-term survival of patients in whom complete eradication of esophageal varices was obtained were compared with those of patients in whom the eradication of varices was incomplete.The frequency of the recurrence/deterioration rate of varices and rebleeding in the complete eradication group was significantly lower than that in the incomplete eradication group (10.7% vs. 100%, 0% vs. 58.3%, respectively, p0.01). Accumulated 5-year survival rate of the complete eradication group was significantly higher than that of the incomplete eradication group (68.2% vs. 43.2%, p0.05).Complete eradication of esophageal varices by endoscopic injection sclerotherapy is effective both in preventing variceal re-bleeding and in improving the survival of cirrhotic patients with esophageal varices.

We report a case of a 61-year-old Japanese woman who developed intra-abdominal angiosarcomatosis 20 years after receiving radiotherapy for squamous cell carcinoma of the cervix.The surgically resected portion of the ileum showed diffuse proliferating angiosarcoma, with irregular channels lined by atypical vascular endothelial cells. Immunohistochemical studies showed that the tumour cells were positive for Factor VIII-related antigen and ulex europaeus agglutinin 1. At autopsy, the tumour had disseminated to the peritoneum and invaded into the right thoracic cavity.These findings were compatible with radiation-induced angiosarcomatosis.

Two cases of axis fracture are reported. Traffic accidents were the cause of injury in both cases. The odontoid process was dislocated anterolaterally with fracture of the lateral mass of the axis in both cases. In Case 1 good alignment of the atlas and axis could not be maintained even with a Halo-vest. The case 2 patient with spinal cord compression caused by dislocation of the axis was referred to our hospital 8 weeks after the accident. Both cases were surgically treated in a similar fashion. Because of the lateral mass fracture and lateral displacement of the atlas relative to the axis, posterior screw fixation with interlaminar fixation (Magerl) was thought to carry a risk of causing damage to the vertebral artery. Because of this, anterior screw fixation combined with posterior wiring was conducted. First, the odontoid process was fixed anteriorly by a screw because it was able to be placed back in good alignment while the patient was in the supine position. Then, the posterior C1-2 interlaminar wiring with iliac bone graft was added. Rigid fixation was obtained without any complication. Various kinds of fixation, such as posterior interlaminar wiring, odontoid screw fixation and Magerl's fixation, have been reported for the treatment of unstable axis fracture. Among them Magerl's method has been regarded as the most stable. When it is not applicable, combination of the first two methods can be an alternative way of treatment for this odontoid process fracture.

Rapid atrial pacing in sinus rhythm may directly induce atrial flutter without provoking intervening atrial fibrillation, or initiate atrial flutter indirectly, by a conversion from an episode of transient atrial fibrillation provoked by rapid atrial pacing. The present study was performed to examine whether or not the direct induction of clockwise or counterclockwise atrial flutter was pacing-site (right or left atrium) dependent.We analyzed the mode of direct induction of atrial flutter by rapid atrial pacing. In 46 patients with a history of atrial flutter, rapid atrial pacing with 3 to 20 stimuli (cycle length = 500 - 170 ms) was performed in sinus rhythm to induce atrial flutter from 3 atrial sites, including the high right atrium, the low lateral right atrium, and the proximal coronary sinus, while recording multiple intracardiac electrograms of the atria. Direct induction of atrial flutter by rapid atrial pacing was a rare phenomenon and was documented only 22 times in 15 patients: 3, 11, and 8 times during stimulation, respectively, from the high right atrium, low lateral right atrium, and the proximal coronary sinus. Counterclockwise atrial flutter (12 times) was more frequently induced with stimulation from the proximal coronary sinus than from the low lateral right atrium (8 vs 1, P = .0001); clockwise atrial flutter (10 times) was induced exclusively from the low lateral right atrium (P = .0001 for low lateral right atrium vs proximal coronary sinus, P = .011 for low lateral right atrium vs high right atrium).Direct induction of either counterclockwise or clockwise atrial flutter was definitively pacing-site dependent; low lateral right atrial pacing induced clockwise, while proximal coronary sinus pacing induced counterclockwise atrial flutter. Anatomic correlation between the flutter circuit and the atrial pacing site may play an important role in the inducibility of counterclockwise or clockwise atrial flutter.

Aurora- and Ipl1-like midbody-associated protein (AIM-1) is a serine/ threonine kinase that is structurally related to Drosophila aurora and Saccharomyces cerevisiae Ipl1, both of which are required for chromosome segregation. A kinase-negative form of AIM-1 inhibits the formation of cleavage furrow without affecting nuclear division, indicating that the gene controls entry into cytokinesis during M phase in mammalian cells. A human gene that encodes the protein AIM-1 was overexpressed in colorectal and other tumor cell lines. The regulation of AIM-1 expression was cell cycle dependent in normal and tumor cells, and the maximum accumulation was observed at G2-M. Exogenous overexpression of wild-type AIM-1 produced multinuclearity in human cells, suggesting that the excess amount of AIM-1 had a dominant-negative effect on the overexpressing cells. In long-term culture of AIM-1-overexpressing cells, multiple nuclei in a cell were occasionally fused, and then an increased ploidy and aneuploidy were induced. Thus, the overexpression of AIM-1 in colorectal tumor cell lines is thought to have a causal relationship with multinuclearity and increased ploidy. Cytokinesis error caused by AIM-1 overexpression is a major factor in the predisposition of tumor cells to the perturbation of chromosomal integrity that is commonly observed in human neoplasia. Thus, defects of pathways essential for mitotic regulation are important during human cancer development.

Human papilloma viruses (HPV) have been regarded seriously not only as an important agent causing sexually transmitted disease, but also because of its association with malignant transformation. Over 70 types of HPV are known, of which 30 types have been detected in specimens from the cervix. These have been classified into a low risk group, a high risk group and an intermediate group according to their association with malignant transformation. Therefore, diagnosis of cervical HPV infection and to a certain extent the HPV type are extremely important. Diagnostic methods of HPV infection include morphological methods such as cytological, pathological and electron microscopical diagnoses; DNA hybridization methods such as Southern blot hybridization (SBH) and Dot blot hybridization (DBH); and a combination of the two methods such as in situ hybridization (ISH), in situ polymerase chain reaction (in situ PCR) and polymerase chain reaction-in situ hybridization (PCR-ISH). Morphological methods using koilocytosis as an indicator have a low detection rate for HPV. Although ISH, in situ PCR and PCR-ISH have lower sensitivity that SBH and DBH, they enable visual localization of the detected DNA in cells or tissues. These methods are therefore important for cytopathologists and histopathologists. In this study, we report the morphological characteristics of HPV, as well as the advantages, disadvantages and detection rates of the various diagnostic methods.

Placental site trophoblastic tumor (PSTT) is a rare form of trophoblastic neoplasm. Approximately 100 cases of PSTT have been reported, but we found no report on its cytodiagnosis.Case 1, a 39-year-old female, came to the hospital because of abnormal genital bleeding. Case 2, a 36-year-old female came because of amenorrhea for a year. In both cases, endometrial smear and intrauterine curettage suggested trophoblastic disease, and hysterectomy was performed. Laboratory data revealed a mild increase in human chorionic gonadotropin (hCG) and beta-hCG but normal human placental lactogen (hPL). In the cytologic examination, the background contained some hemorrhagic and fibrinous areas but no necrosis. Most tumor cells stained light green, were round or polygonal, and contained abundant cytoplasm. Some were palely stained and had vacuoles. Some cells showed hyperchromatism, an irregular nucleus, fine-to-coarse chromatin granules and markedly different sizes. Most of the cells were hPL positive, and a few were hCG positive.Both cases were considered benign because of rare mitoses despite cellular pleomorphism. However, careful follow-up is required. The differential diagnosis of PSTT is difficult from cytologic and biopsy specimens alone but may be achieved with additional magnetic resonance imaging findings and positive staining of hPL and hCG.

Angiogenesis is critical for tumor growth and inflammation. Shiunko is a Chinese herbal ointment used for the treatment of burns in Japan. Its main ingredient is the root of Lithospermum erythrorhizon, which had been used for treating tumors and inflammation in China since the 5th century. We report here that shikonin, the main chemical ingredient of L. erythrorhizon is a novel inhibitor of angiogenesis. It inhibited tumor necrosis factor-alpha-induced and B16 melanoma-induced angiogenesis in mice and normal developmental angiogenesis in the yolk-sac membranes of chick embryos. Shikonin also inhibited proliferation and migration of endothelial cells in culture and network formation by endothelial cells on Matrigel in vitro. The dose-responsive study suggests that the mechanism of this inhibitory effect on angiogenesis involves the prevention of network formation by endothelial cells via blocking integrin alpha v beta 3 expression.

Endodermal sinus tumor (EST) of vaginal origin is extremely rare, and few previously published reports have described the cytology of this tumor.A high serum alpha-fetoprotein (AFP) level and magnetic resonance imaging findings in a 10-year-old female suggested EST of the lower genital tract. Laparotomy revealed EST occupying the pelvic cavity. The patient died on the 17th postoperative day due to complications from disseminated intravascular coagulation. Autopsy was performed immediately after death. The tumor originated in the posterior vaginal wall directly below the ectocervical area. Both ovaries were involved, and the tumor proliferated to occupy the cavity of the pelvis minor. A vaginal smear, obtained on two occasions, revealed severe necrosis with few cells, and a diagnosis could not be made. An imprint smear of the tumor obtained during surgery showed relatively characteristic cytologic findings of EST. Immunocytochemical and immunohistochemical studies revealed staining positive for AFP.Vaginal EST usually occurs in patients under 3 years of age. This 10-year-old patient was the oldest among the reported cases. Vaginal EST is clinically similar to sarcoma botryoides and clear cell carcinoma, and a differential diagnosis is necessary. Immunocytostaining for AFP is essential for a definitive cytologic diagnosis of EST.

The 11C-labeled KF17837 ([7-methyl-11C](E)-8-(3,4-dimethoxystyryl)-1,3-dipropyl-7-methylxa nthine) was evaluated as a PET ligand for mapping adenosine A2a receptors in the central nervous system (CNS).The regional brain distribution of [11C]KF17837 and the effect of adenosine antagonists on the distribution were measured in mice by the tissue sampling method. In rats, the regional brain uptake of [11C]KF17837 and the effect of carrier KF17837 was visualized by autoradiography. Imaging of the monkey brain with [11C]KF17837 was performed by PET.In mice, a high uptake of [11C]KF17837 was found in the striatum in which A2a receptors were highly enriched. The uptake was decreased by co-injection of carrier KF17837 or a xanthine-type A2a antagonist CSC but not by nonxanthine-type A2a antagonists ZM 241385 or SCH 58261, or an A1 antagonist KF15372. In the rat brain, [11C]KF17837 was accumulated higher in the striatum than in other brain regions, and the uptake was blocked by co-injection of carrier KF17837. In a monkey PET study, a high striatal uptake of radioactivity was observed.Carbon-11-KF17837 binds to adenosine A2a receptors in the striatum. However, the presence of an unknown but specific binding site for xanthine-type compounds also was suggested in the other brain regions. The results also suggested that the in vivo receptor-binding sites of xanthine-type ligands are slightly different from those of nonxanthine-type A2a antagonists.

Cartilage is unique among tissues of mesenchymal origin in that it is resistant to vascular invasion due to an intrinsic angiogenic inhibitor. During endochondral bone formation, however, calcified cartilage formed in the center of the cartilaginous bone rudiment allows vascular invasion, which initiates the replacement of cartilage by bone. The transition of cartilage from the angioresistant to the angiogenic status thus plays a key role in bone formation. However, the molecular basis of this phenotypic transition of cartilage has been obscure. We report here purification of an endothelial cell growth inhibitor from a guanidine extract of bovine epiphyseal cartilage. The N-terminal amino acid sequence indicated that the inhibitor was identical to chondromodulin I (ChM-I), a cartilage-specific growth-modulating factor. Purified ChM-I inhibited DNA synthesis and proliferation of vascular endothelial cells as well as tube morphogenesis in vitro. Expression of ChM-I cDNA in COS7 cells indicated that mature ChM-I molecules were secreted from the cells after post-translational modifications and cleavage from the transmembrane precursor at the predicted processing signal. Recombinant ChM-I stimulated DNA synthesis and proteoglycan synthesis of cultured growth plate chondrocytes, but inhibited tube morphogenesis of endothelial cells. In situ hybridization and immunohistochemical studies indicated that ChM-I is specifically expressed in the avascular zone of cartilage in developing bone, but not present in calcifying cartilage. These results suggest a regulatory role of ChM-I in vascular invasion during endochondral bone formation.

A series of 8-substituted derivatives of 3,7-dimethyl-1-propargylxanthine (DMPX) was synthesized and investigated as A2A adenosine receptor antagonists. Different synthetic strategies for the preparation of DMPX derivatives and analogues were explored. A recently developed synthetic procedure starting from 3-propargyl-5,6-diaminouracil proved to be the method of choice for the preparation of this type of xanthine derivatives. The novel compounds were investigated in radioligand binding studies at the high-affinity adenosine receptor subtypes A1 and A2A and compared with standard A2A adenosine receptor antagonists. Structure-activity relationships were analyzed in detail. 8-Styryl-substituted DMPX derivatives were identified that exhibit high affinity and selectivity for A2A adenosine receptors, including 8-(m-chlorostyryl)-DMPX (CS-DMPX, Ki A2A = 13 nM, 100-fold selective), 8-(m-bromostyryl)-DMPX (BS-DMPX, Ki A2A = 8 nM, 146-fold selective), and 8-(3,4-dimethoxystyryl)-DMPX (Ki A2A = 15 nM, 167-fold selective). These and other novel compounds are superior to the standard A2A adenosine receptor antagonists KF17837 (4) and CSC (5) with respect to A2A affinity and/or selectivity.

To evaluate the mechanism of localized portal hypertension associated with stenosis or obstruction of the splenic vein in chronic alcoholic pancreatitis.Surgical and autopsy specimens from 12 patients with clinically diagnosed chronic alcoholic pancreatitis were examined histopathologically. Autopsy specimens of 10 normal pancreases served as control tissues.In tissues from 11 of the 12 patients with chronic-alcoholic pancreatitis, fibrosis in the pancreatic parenchyma continuously extended to the wall of the splenic vein; organized thrombus formation with recanalization was found in five patients, phlebosclerosis in four, and no changes were found in two. In two of the five patients with organized thrombus formation in the splenic vein, localized portal hypertension had been clinically diagnosed because of splenomegaly and varicose veins in the fundus of the stomach, but it was not accompanied by liver cirrhosis. In the control tissues, fibrosis was not observed in the peripancreatic tissue or the area surrounding the wall of the splenic vein.We regard localized portal hypertension due to stenosis or obstruction of the splenic vein as one of the consequences of peripancreatic fibrosis in chronic alcoholic pancreatitis.

We have described previously that CD8+ CD11b+ TCR gamma/delta+ type 2 T cells (BA-type 2 T cells) and suppressor macrophages (Sup-Mł) are generated in spleens of mice 3 to 5 days (Sup-Mł) and 4 to 9 days (BA-type 2 T cells) after thermal injury. In the present study, an additional suppressor T cell, characterized as CD3+ CD4- CD8- TCR alpha/beta+ T cells (double negative suppressor T cells, DN Sup-T cells), was demonstrated in mice 1 day after thermal injury. DN Sup-T cells inhibited the proliferation of lymphocytes stimulated with allogeneic cells or a lectin in a mixed lymphocyte reaction, and produced both type 1 and type 2 cytokines (interferon-gamma, interleukin-2, interleukin-4, and interleukin-10) when they were stimulated in vitro with anti-CD3 monoclonal antibody. These results suggest that DN Sup-T cells express phenotypic properties similar to natural suppressor cells and cytokine-producing profiles different from type 1 (Th1 cells and CTLs) and type 2 T cells (Th2 cells and CD8+ type 2 T cells). DN Sup-T cells may play a role on the burn-associated immunosuppression appearing in the acute phase of thermally injured individuals.

Keishi-ka-kei-to, a traditional Chinese herbal medicine composed of a mixture of crude extracts from five medicinal plants (Cinnamomi cortex, Paeoniae radix, Zizyphi fructus, Zingiberis rhizoma and Glycyrrhizae radix), inhibited experimental pulmonary metastasis in mice implanted with B16F10 melanoma cells. When 136 to 145 metastatic colonies were produced in lungs of mice inoculated with 1 x 10(5) cells/mouse of melanoma cells, less than 15 metastatic colonies were demonstrated when these tumor-inoculated mice were treated orally with 80 to 320 mg/kg doses of Keishi-ka-kei-to. The most active component in the mixture was shown to be 6-gingerol, derived from the Zingiberis rhizoma extract. The antimetastatic activity of 6-gingerol was expressed through the host's antitumor immune functions, as the growth of B16F10 melanoma cells was not affected by this substance in vitro. The splenic CD8+ T cells from mice treated with the compound showed inhibitory activities on pulmonary metastasis when these T cells were adoptively transferred to mice bearing B16F10 melanoma cells. These results may suggest that Keishi-ka-kei-to inhibits pulmonary metastasis in mice bearing B16F10 melanoma cells through the stimulation of CD8+ T cells.

A cDNA clone, which has an open reading frame of 1398 nucleotides encoding a 466-amino-acid peptide, has been isolated from rabbit liver cDNA library. Compared with the peptide sequence, it shows high homology to alpha1a adrenoceptors of human, bovine and rat. We expressed this clone in COS-7 and investigated the pharmacological properties, revealing similarity to those of human alpha1a adrenoceptors. Competitive RT/PCR has detected the mRNA in variety of rabbit tissues, especially abundantly in liver, vas deferens, brain, and aorta, but not in heart.

This study was carried out to determine the relationship of conduction delay during pace-mapping, as indicated by the stimulus to QRS interval to different reentry circuit sites and bystanders, as identified by entrainment criteria, in patients with ventricular tachycardia late after myocardial infarction. Catheter mapping and ablation were performed in 40 patients with ventricular tachycardia after myocardial infarction. Data were retrospectively analyzed from 122 endocardial sites which met the following the criteria: a 12-lead electrocardiogram was recorded during pace-mapping, ventricular tachycardia was then induced and entrained by pacing to classify the type of the site relative to the reentry circuit exit. By entrainment criteria 77 sites were in the reentry circuit (28 exit sites, 49 other circuit sites) and 45 sites were bystanders not in the reentry circuit. The average stimulus conduction delay at central/proximal sites was 103 +/- 43 ms, which was significantly longer than at exit (57 +/- 31 ms), outer loop sites (57 +/- 32 ms), and bystander sites (57 +/- 37 ms); P0.05. Pace-mapping revealed evidence of greater conduction delay at sites proximal to the reentry circuit exit, consistent with slow conduction between these regions and the border of the infarct. Exit sites and outer loop sites were more likely to be located along the border of the infarct, with less conduction delay evident during pace-mapping.

The carbon-11-labeled selective adenosine A1 antagonist KF15372 ([1-propyl-11C]8-dicyclopropylmethyl-1,3-dipropylxanthine) was elevated in vivo as a PET ligand for mapping CNS adenosine A1 receptors.The regional brain distribution of [11C]KF15372 and the effects of adenosine antagonists on the distribution were determined in mice by tissue sampling. In rats, in which the retinal projection fibres to the superior colliculus had degenerated due to unilateral eye removal, the brain distribution of [11C]KF15372 was visualized by ex vivo autoradiography.The mouse brain uptake of [11C]KF15372 was 1.8% i.d./g at 5 min and then it gradually decreased. The uptake was high in the hippocampus, cerebral cortex, striatum and cerebellum, and was significantly reduced by A1 antagonists but not by A2 antagonists. The brain distribution of 11C assessed by the tissue sampling and autoradiography was compatible with that of the A1 receptors. Autoradiography clearly visualized unilaterally decreased A1 receptor binding in the superior colliculus.The results demonstrated that [11C]KF15372 is a selective and high-affinity adenosine A1 receptor ligand and is useful for detecting the degeneration of presynaptic neurons.

Adenosine produces tone-dependent pulmonary vascular responses; however, the adenosine receptor subtype mediating these responses is unknown. In the present study, the adenosine receptor subtypes mediating tone-dependent responses were investigated, Intralobar injections of adenosine,ATP, and analogues under low-tone conditions caused dose-related increases in lobar arterial pressure; the order of potency was alpha,beta-methylene ATP (alpha,beta-metATP) > N6-cyclopentyladenosine (CPA) > ATP > adenosine. Under low-tone conditions, pressor responses to adenosine, ATP, and CPA, an adenosine A1-receptor agonist, were reduced by KW-3902, an adenosine A1-receptor antagonist, whereas KW-3902 and meclofenamate had no effect on responses to alpha,beta-metATP, norepinephrine, serotonin, or angiotensin II. Under elevated-tone conditions, injections of adenosine, ATP, and analogues caused dose-related decreases in lobar arterial pressure, and adenosine was 10-fold less potent than 5'-(N-cyclopropyl)-carboxamidoadenosine (CPCA), an A2-receptor agonist, and ATP. KF-17837, an A2-receptor antagonist, reduced vasodilator responses to adenosine and CPCA, whereas responses to ATP, isoproterenol, diethylamine-NO, lemakalim, and bradykinin were not changed. The vasodilator responses to adenosine were not attenuated by Nw-nitro-L-arginine benzyl ester, methylene blue, or U-37883A. These results suggest that vasoconstrictor responses to adenosine are mediated by A1 receptors and the release of vasoconstrictor prostanoids, and that, under elevated-tone conditions, vasodilator responses are mediated by A2 receptors but not the release of nitric oxide or the activation of guanylate cyclase or K+ATP channels.

This study determines, in vivo, whether endogenous adenosine/A1 receptor interactions at juxtaglomerular cells restrain the release of renin induced by receptor-mediated activation of the adenosine 3',5'–cyclic monophosphate pathway and whether endogenous adenosine/A2 receptor interactions diminish this restraining response. The following four pharmacological probes were employed: 1) 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) and 2) FK-453, both selective A1 receptor antagonists; 3) FR-113452, a nearly inactive enantiomer of FK-453; and 4) KF-17837, a selective A2 receptor blocker. Adult Sprague-Dawley rats were prepared (adrenalectomized, renal denervated, uninephrectomized, and treated with indomethacin, aldosterone, and hydrocortisone) to minimize endogenous stimulation of renin release and received either vehicle (control group) or one of the four drugs. Intrarenal infusions of isoproterenol (3, 30, and 100 ng.kg-1.min-1) caused dose-related increases in plasma renin activity (PRA). This PRA response was significantly augmented in the groups receiving DPCPX (P = 0.0010) or FK-453 (P = 0.0001) but was not altered in the groups treated with FR-113452 (P = 0.3422) or KF-17837 (P = 0.2155). Systemic and renal hemodynamics and renal electrolyte excretions were monitored and could not account for the PRA augmentation caused by the A1 antagonists. This study clearly demonstrates that endogenous adenosine acts on the A1 receptor to restrain the renin release induced by activation of intrarenal beta-adrenoceptors and is not counteracted by endogenous activation of the A2 receptor.

Using immuno-colloidal gold surface replicas (IGSR), we developed a method to demonstrate surface antigens in a whole view of a cell at the EM level. In this study, a monoclonal antibody (MAB) 7C5 directed against sp56, which is a mouse sperm protein (M(r) = 56,000 daltons) and has specific affinity for the zona pellucida, was used as primary antibody. Fixed mouse spermatozoa were incubated with primary antibody and colloidal gold-labeled secondary antibody on a slide glass. Used gold particles were 10, 20 or 30 nm in diameter. The spermatozoa were then critical point dried, and replicated with platinum and carbon. Replicas were examined with a transmission EM. The result suggests that sp56 is located on the outer surface of the plasma membrane covering approximately 2/3 of dorsal edge of the sperm head, but it is virtually absent in the tapered apical tip. When 10 and 20 nm colloidal particles were used, about 500 particles were observed on one side of the flat sperm head. Although the large 30 nm particles were easily recognized in low magnification figures, the attached number on the sperm head was generally lower than the number of 10 or 20 nm particles observed. By this method, we could effectively demonstrate the localization of a surface antigen in a wide cell view with clear landmarks.

Intraperitoneal or intrahippocampal injections of kainate induce both hippocampal cell death and axonal remodeling of the dentate gyrus granular neurons. We report here that injection of kainate into the dorsal hippocampus of adult mice may also trigger a conspicuous and long-lasting global trophic response of granule cells. Morphological changes include somatic and dendritic growth and increased nuclear volume with ultrastructural features characteristic of neuronal development. The trophic response is correlated with a specific overexpression of brain-derived neurotrophic factor that is maintained for at least six months. This shows that plasticity in adult neurons can, in addition to axonal remodeling, extend to generalized cell growth. Our results further suggest that brain-derived neurotrophic factor could be involved in the activation and/or maintenance of this phenomenon.

Human papilloma virus (HPV) infection is diagnosed by morphological methods (including cytodiagnosis, histodiagnosis and electron microscopy) and DNA diagnostic methods. One of the DNA diagnostic methods, Dot blot hybridization (DBH), is generally regarded to be superior to the morphological methods in terms of specificity and sensitivity. There have recently been reports showing that such morphological methods as cytodiagnosis [for koilocytotic atypia (KC) or koilocytosis] and histodiagnosis are best for HPV detection in laboratories because they have better sensitivity, and are cheaper, and procedurally simpler than the DBH method. We compared detection rates by morphological methods (cytodiagnosis and histodiagnosis) and the DBH method. (Materials and Methods) The subjects were 377 patients who came to the Department of Obstetrics and Gynecology of Juntendo University Urayasu Hospital and had abnormal findings in the uterine cervix by cytodiagnosis of colposcopy. Uterine cervix swabs were scraped during colposcopic examination, and specimens for Papanicolaou and Giemsa staining were prepared on 2 slides. Scraping was again performed for the DBH (Vira pap. and type, Toray Industries, Inc.) method using swabs specified for the kit, and cells were obtained in a collection kit. At the same time, at least 3 sites including, possible atypical conversion zones, were biopsied, followed by paraffin embedding and hematoxylin-eosin staining of the specimens. (Results) 1) Comparison of detection rates by cytodiagnosis and the DBH method The positive rates by the DBH method were determined according to cytodiagnostic category in 377 patients. HPV-DNA was detected in every category and in 86 (22.8%) of all of the patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Transgenic mice have recently been used for mutagenesis assays in vivo. The present study was undertaken to clarify whether such assays can detect mutations induced after treatment of male germ cells in mouse with isopropyl methanesulfonate (iPMS), ethylnitrosourea (ENU) or X-ray irradiation. The transgenic mice used for assay are Muta Mouse (MM) strain, which carries 80 copies of the bacterial lacZ gene per cell as targets for mutagenesis. Male MM animals were given a single intraperitoneal injection of 200 mg/kg iPMS, 150 mg/kg ENU or were irradiated with 500 rads of X-rays. Vasa deferential sperm, caudal epididymal sperm and/or whole testes were extracted at various times after treatment with each agent. After the genomic DNA was extracted from each tissue, mutation analysis at the lacZ locus was carried out by the method of Myhr et al. The spontaneous lacZ- mutant frequencies were on the order of 10(-5)-10(-6). The lacZ- mutant frequencies in all treatment groups were increased over the control animals. The iPMS-induced mutant frequency in postmeiotic stages was low. However, ENU induced relatively high mutant frequencies in the spermatogonia. X-rays induced mutant frequencies in the late spermatid and early spermatid stages that were higher than the mutant frequencies in spermatogonia. Mutant frequencies in MM detected after treatment of male germ cells with ENU or X-rays were lower than mutant frequencies detected by the mouse specific-locus test in previous reports. Hence, considering the lower resolution power of the transgenic animal mutagenesis assays using the target lacZ gene compared with the specific locus test, to detect mutations induced in male germ cells, it is not clear whether this assay is a practical alternative to the specific locus test.

Changes of liver functions associated with endoscopic injection sclerotherapy (EIS) in which less than 6 ml of 5% ethanolamin oleate was used were investigated in 50 patients with remarkable esophageal varices. The index used for evaluating hepatic reserve was the difference of total bilirubin in serum between immediately and 15 hours after EIS and was tentatively named as "delta TB" (normal rangeor = 0.1). In the patients classified as Child-Pugh A (n = 25), Child-Pugh B (n = 19) and Child-Pugh C (n = 6), delta TB was 0.04 +/- 0.30, 0.23 +/- 0.41, and 0.54 +/- 0.21, respectively. One month after the entire sessions of EIS, the changes of liver function tests before and after EIS were compared in 45 cases. It was disclosed that delta TB was elevated in parallel with the severity of their liver dysfunction before EIS. Also, delta TB seems to reflect the prognosis of the patients treated by EIS for some extent, since the survival period of the cases treated by EIS were correlative with delta TB (r = -0.734, p0.01), and delta TB of 20 cases who survived for longer than 5 years after EIS was 0.04 +/- 0.32. Therefore, delta TB seems to be a useful marker for estimating the influence of EIS on liver function.

Endoscopic intravariceal injection sclerotherapy (EIS) using sclerosant mixed with contrast medium was done in 100 patients without hepatocellular carcinoma. They were followed longer than 12 months (mean; 58 +/- 29 months) after EIS. The recurrence rate of esophageal varices in cases with complete eradication (n = 79) and cases with incomplete eradication of (n = 21) was 8.9% and 85.7%, respectively (p0.01). In 21 cases, complete eradication was achieved by intravariceal injection and additional therapy using paravariceal injection was not performed. The recurrence rate of this group was only 4.8%. Endoscopic varicealography during injection sclerotherapy were evaluated in 91 cases. At final session of EIS, narrowing of diameter (less than 1 mm) and irregularity of vessel walls were observed in small vessels (devastated vessels). Appearance rate of devastated vessel in 75 cases with completely eradicated esophageal varices was 65.3%. In contrast, among 16 cases with incomplete eradication of varices, devastated vessels were seen only in 6.3% (p0.01). It is concluded that the important point in preventing the recurrence of esophageal varices after EIS was the complete eradication of esophageal varices by intravariceal injection sclerotherapy resulting in the eradication of the routes to esophageal varices from port-splenic venous system. For the sake of accomplishment of this treatment, appearance of devastated vessel is very useful.

Rabbit chondrocytes were isolated from resting cartilage of ribs of 4-week-old New Zealand rabbits. Chondrocytes were seeded at low density and grown to confluency in medium (DME) with 10% fetal bovine serum, 50 micrograms/ml ascorbic acid, and antibiotics, at 37 degrees C under 5% CO2 in air, and then serum concentration was reduced to 0.3%. At the low serum concentration, chondrocytes adopted fibroblastic morphology. Addition of concanavalin A to the culture medium induced a morphologic alteration of the fibroblastic cells to polygonal or spherical chondrocytes that were surrounded by refractile matrix. Wheat germ agglutinin and garden pea lectin induced similar cell shape changes without any increase in matrix synthesis. Concanavalin A decreased [3H]thymidine incorporation into DNA in a dose-dependent manner with an ED50 of 0.4-1 micrograms/ml in the presence of various concentrations of serum ranging from 0.3 to 20%. The concanavalin A inhibition of DNA synthesis was abolished by 10 mmol/L methyl-D-mannopyranoside. The inhibition of chondrocyte proliferation does not seem to be specific to concanavalin A. Wheat germ agglutinin, lentil lectin, phytohemagglutinin, Ulex europeaus agglutinin, and garden pea lectin also decreased, dose-dependently, [3H]thymidine incorporation into DNA in chondrocytes. Other lectins at 0.01-20 micrograms/ml had little effect on [3H] thymidine incorporation. Because the molecular structure of concanavalin A and its mode of actions have been extensively characterized, chondrocytes exposed to this lectin will be useful as a novel model in studying of the control of cellular differentiation.

Nephrocalcin, an acidic glycoprotein that inhibits calcium oxalate crystal growth, has been previously localized in proximal tubules of kidneys by an immunohistochemical staining method and purified from tissue culture media of 2 renal carcinoma cell lines. A polyclonal antibody specific to nephrocalcin was raised in rabbits and the level of nephrocalcin was quantitatively determined in urine of 19 renal cell carcinoma patients (0.241 +/- 0.341 microgram nephrocalcin per mg. creatinine) and compared to healthy controls (0.022 +/- 0.012 micrograms nephrocalcin per mg. creatinine). Nephrocalcin levels after tumor nephrectomy decreased dramatically in 5 patients and to a lesser degree in 7. A specific nephrocalcin fraction that was eluted from an anion exchange column with low ionic strength was detected in urine of the renal cell carcinoma patients, and this fraction decreased or disappeared after tumor nephrectomy in 6 of 9 patients studied. Amino acid composition, phosphate content and dissociation constants toward calcium oxalate monohydrate crystals were investigated in the nephrocalcin from tumor patients and compared to that from healthy controls. Our studies demonstrate that nephrocalcin in patients with renal cell carcinoma is atypical and usually in much higher quantity. Further studies are needed to determine the clinical significance of these observations.

The enhanced expression of the trophic factor transforming growth factor alpha (TGF alpha) in reactive astrocytes following CNS injury suggests that TGF alpha has a role in the development of astrogliosis. We explored this hypothesis in the murine mutant wobbler, which presents a progressive motoneuronal degeneration associated with an astrogliosis. Evolution of astrogliosis, and expression of TGF alpha precursor (pro-TGF alpha) and of its receptor were examined over the course of the disease, using genetically diagnosed animals and immunocytochemical techniques. We report here that degenerating motoneurons of the cervical spinal cord and a subset of astrocytes express pro-TGF alpha, prior to the onset of astrogliosis, when the first clinical manifestations of the disease are observed at 4 weeks of age. TGF alpha expression appeared strongly correlated with motoneuronal degeneration. All pro-TGF alpha-immunoreactive neurons exhibited a degenerative morphology, and the number of pro-TGF alpha- immunoreactive neurons increased with the progression of the disease. At the glial level, we observed that astrogliosis was a transitory phenomenon in the wobbler mice, developing in coordination with the motoneuronal expression of pro-TGF alpha. Astrogliosis became evident in 6-week-old wobbler mice, when the number of pro-TGF alpha- immunoreactive motoneurons was maximal, and regressed in older mutant mice in correlation with the disappearance of pro-TGF alpha- immunoreactive motoneurons. Furthermore, TGF alpha/EGF receptor immunoreactivity was exclusively localized in a subset of reactive astrocytes, its expression following closely the course of the astrogliosis. These data show that TGF alpha synthesis by the affected motoneurons is an early event in the course of the wobbler disease, and suggest a role for TGF alpha as a neuronal inducer of astrocytic reactivity.

The purpose of this study was to investigate the relationship between changes in parathyroid hormone (PTH) receptor levels and chondrocyte maturation during endochondral ossification. Chondrocytes were isolated from the growth plate of rabbit ribs and maintained in the presence of 10% serum in mass cultures. Treatment with PTH-(1-84) and a PTH-(1-34) fragment suppressed the increases in alkaline phosphatase activity and in type X collagen and 1 alpha,25-dihydroxyvitamin D3 receptor levels and abolished 45Ca incorporation into mineral, all of which occurred in parallel untreated cultures in the hypertrophic (terminal) stage. These effects of PTH were observed at low concentrations (10(-10) to 10(-9) M) and within 24-48 h of treatment. PTH-(1-84) and PTH-(1-34) also increased [35S]sulfate incorporation into newly synthesized proteoglycans. In contrast, the middle and carboxyl-terminal fragments of PTH tested had little effect on proteoglycan synthesis or terminal differentiation. The binding of 125I-PTH-(1-34) to cells in the growth plate was greater than that to cells in liver, skin, muscle, brain, or kidney. When the correlation between binding levels and stage of maturation was examined, we found that 125I-PTH-(1-34) binding to its 72-kDa receptor was low in resting and proliferating chondrocytes, increased 10-fold in matrix-forming chondrocytes, and thereafter decreased in hypertrophic chondrocytes both in vitro and in situ. Scatchard analysis revealed that the changes in PTH binding were due to changes in the number, and not in the affinity, of the receptor. The changes in PTH-(1-34) binding paralleled those in [35S]sulfate incorporation into proteoglycans. These findings suggest that stage-dependent increases in PTH/PTH-related peptide receptor levels localize the hormone stimulation of proteoglycan synthesis and inhibition of precocious hypertrophy in the matrix-forming zone of growth plates.

Changes of liver functions associated with endoscopic injection sclerotherapy (EIS) were investigated in 143 patients with remarkable esophageal varices. The index used for evaluating hepatic reserve was the ratio between the increase of total bilirubin and the increase of LDH after EIS [I.I. index = (delta T.Bil./delta LDH) x 100] in cases treated by intravariceal injection of 5% ethanolamine oleate. I.I. index value caused by hemolysis was stable and always below 0.2, while the elevation of I.I. index above 0.2 was regarded as the reflection of the deterioration of liver function. After the entire sessions of EIS, the changes of liver function tests before and after EIS were estimated in 104 cases using 3 factors as follows; (1) delta T.Bil.; increase of total bilirubin more than 0.5 mg/dl (2) delta PT; decrease of prothrombin time more than 5% (3) delta ICG R15; increase of ICG R15 more than 5%. I.I. index of group I (without any changes of 3 factors mentioned above; n = 66) was 0.22 +/- 0.16, approximately within normal range. However, in group II (with 1 factor; n = 29), group III (with 2 factors; n = 5), group IV (with all factors; n = 4), I.I. index was elevated according to the severity of their liver dysfunction. The cases whose I.I. index value increased gradually during the course of EIS, were apt to deteriorate the liver functions and therefore, should be carefully observed. Also, I.I. index seems to reflect the prognosis of the patients treated by EIS for some extent, since the survival period of the cases treated by EIS were correlative with the value of I.I. index (r = -0.542, p0.01), and I.I. index of 32 cases who survived for longer than 5 years after EIS was almost normal (0.22 +/- 0.15).

The case was in a 7-month-old boy. Rubinstein-Taybi syndrome was diagnosed from the broad halluces and the characteristic physiognomy noticed since birth. He was brought to us at 7 months of age with fever as the chief complaint. Voiding cystourethrogram showed bilateral vesicoureteral reflux. The testis was palpable in the inguinal canal on either side, but could not be brought to the scrotum. No abnormalities were detected in the penis. From the above findings, he was diagnosed to have bilateral 4th degree vesicoureteral reflux and undescended testes as complicating Rubinstein-Taybi syndrome. The condition was treated by bilateral anti-reflux repair and bilateral orchidopexy at 8 months of age. There has been no recurrence of vesicoureteral reflux for 3 years. A brief discussion was made about the urological complications of Rubinstein-Taybi syndrome.

The goal of this study was to determine whether (E)-1,3-dipropyl-7-methyl-8-(3,4-dimethoxystyryl)xanthine (KF17837) is a useful pharmacological probe for investigating in the rat the in vivo physiological roles of A2 adenosine receptors. In anesthetized rats, bradycardic responses to N6-cyclopentyladenosine and hypotensive responses to 2-[p-(2-carboxyethyl)phenethylamino]-5'-N-ethylcarboxamido adenosine (CGS21680C) were used to assess A1 receptor and A2 receptor activation, respectively. After obtaining control responses to N6-cyclopentyladenosine and CGS21680C, the rats received infusions of either vehicle or one of two dosage levels of KF17837, a compound recently demonstrated to be a potent and selective A2 receptor antagonist in vitro. KF17837 was infused for 4 hr and, at various times during the infusions, bradycardic and hypotensive responses to N6-cyclopentyladenosine and CGS21680C, respectively, were elicited. Infusion of either 10 or 30 micrograms kg-1 min-1 (2.4 or 7.4 mg kg-1 4 hr-1) of KF17837 did not significantly affect the bradycardic responses to N6-cyclopentyladenosine. By contrast, 10 micrograms kg-1 min-1 of KF17837 attenuated and 30 micrograms kg-1 min-1 of KF17837 nearly abolished hypotensive responses to CGS21680C. In a second study, pretreatment with KF17837 (30 micrograms kg-1 min-1) did not affect the hypotensive response to either PGI2 (3 micrograms kg-1 min-1) or acetylcholine (100 micrograms kg-1 min-1); however, it attenuated the hypotensive response to adenosine (300 micrograms kg-1 min-1). In a third study, hypotension was induced and maintained with an infusion of adenosine (300 micrograms kg-1 min-1).(ABSTRACT TRUNCATED AT 250 WORDS)

A case of non-Hodgkin's T cell lymphoma (diffuse lymphoma, large cell type) associated with marked eosinophilia and pleurisy in a 57-year-old male is reported. The leukocyte count was 12.5 x 10(3)/microliters and eosinophil count was 53% and the absolute count of 6.6 x 10(3)/microliters. The patient's serum and pleural effusion fluid, containing abundant lymphoma cells, showed eosinophil colony stimulating factor (Eo-CSF) activity. Conditioned medium (CM) prepared from patient's T cells (T-CM) produced Eo-CSF and this was enhanced by interleukin-2 (IL-2) stimulation. We demonstrated that the patient's serum contained a significant amount of interleukin-5 (IL-5) and the patient's T-CM, particularly after IL-2 stimulation contained a significant amount of granulocyte-macrophage colony-stimulating factor (GM-CSF). These findings suggest that Eo-CSF produced by neoplastic T cells or normal T cells activated by tumor antigen stimulated the production of eosinophils in this patient and that both IL-5 and GM-CSF might play a role in Eo-CSF activity.

The effect of Z-100, a natural immunomodulator extracted from Mycobacterium tuberculosis strain Aoyama B, on concomitant antitumor immunity (CAI) was investigated in mice immunized with Meth-A tumor cells (primary inoculation) in combination with Corynebacterium parvum. CAI, which was observed in mice 10 days after immunization (I10 mice), disappeared in mice 20 days after immunization (I20 mice). However, no growth of secondary inoculated Meth-A tumors was demonstrated in I20 mice treated with Z-100 (IZ20 mice). CAI was demonstrated in tumor-bearing recipients when splenic mononuclear cells (SMNC) from I10 mice or IZ20 mice were transferred to recipients intralesionally. However, CAI was not detected when recipients received SMNC from I20 mice or a SMNC mixture from I10 and I20 mice. The SMNC mixture from I10 and IZ20 mice inhibited the growth of Meth-A solid tumors in the recipients. The activity of non-specific suppressor cells, which were characterized as CD8+ T cells, was demonstrated in SMNC from I20 mice, while SMNC from I10 and IZ20 mice did not show any suppressor cell activities. In addition, clear inhibition of tumor growth was demonstrated in recipient mice which received a SMNC mixture from I20 mice, previously treated with anti-Lyt-2+ MAb plus complement, and I10 mice. These results suggest that Z-100 might be able to prolong CAI observed in I10 mice through the inhibition of Lyt-2+ T suppressor cells detected in SMNC from I20 mice.

Acute scrotum is a clinical syndrome which includes various disorders. Differential diagnosis of these disorders is not always easy. We reviewed cases experienced in our hospital, focusing on diagnostic procedures. Final diagnosis in 39 cases were acute epididymitis in 23 cases, torsion of the testis in 13 cases, torsion of appendix epididymis in 2 cases and torsion of appendix testis in 1 case. It became clear ultrasonography was a useful diagnostic procedure; if the torsed portion is delineated, torsion of the testis can be diagnosed. Heterogenous echoes from the interior of the testis indicate the necrosis of the testis. Inspection was also useful; if the blue dot sign is detected, torsion of the appendix testis/epididymis is indicated. However, Doppler ultrasound did not always provide a correct diagnosis of torsion of the testis. If torsion of the spermatic cord cannot be completely ruled out, early surgical intervention and treatment are considered essential.

A series of 4-hydroxy-3-quinolinecarboxylic acid derivatives (6) and 4-hydroxy-2-oxo-1,2-dihydro-3-quinolinecarboxylic acid derivatives (7) were designed and synthesized as 5-HT3 receptor antagonists. Molecular modeling studies suggested that the 3-carbonyl moiety in 6 was almost coplanar to the plane of an aromatic ring, but in 7 there was a 30 degrees deviation. 4-Hydroxy substitution in quinoline derivatives enhanced affinity for the 5-HT3 receptors, and endo-N-(8-methyl-8-azabicyclo[3.2.1]oct-3-yl)-4-hydroxy-3- quinolinecarboxamide (6f) exhibited the most potent activity in the Bezold-Jarisch (B-J) reflex test (ED50 = 0.1 micrograms/kg, iv) among quinoline derivatives 6. Although 4-hydroxy-2-oxo-1,2-dihydro-3-quinolinecarboxamide derivatives (7a) exhibited higher affinity (e.g., 7d: Ki = 0.48 nM) for the 5-HT3 receptors than ondansetron (Ki = 7.6 nM) or granisetron (Ki = 2.1 nM), these amides showed less potent activity in the B-J reflex test than the reference compounds. Interestingly, the ester derivatives 7c, 7f, and 7h eliminated affinity for the 5-HT3 receptors. These unusual structure-activity relationships and the deviation of the 3-carbonyl moiety from the plane of an aromatic ring suggest that the active conformation of 7a might be different from the proposed one for the preceding 5-HT3 antagonists. Thus, 6f was chosen for further studies. No receptor binding for a variety of ligands was significantly antagonized by 6f. Comparing the ratios of the ED50 value in the B-J reflex test (rat, iv) with the LD50 value in acute lethal toxicity (mouse, iv), 6f was proved to have a 600-fold wider margin of safety than ondansetron. Compound 6f dose-dependently attenuated both the incidence and frequency of emetic episodes induced by cisplatin in the dog (ED50 = 14 micrograms/kg, iv) more potently than ondansetron (ED50 = 210 micrograms/kg, iv). Compound 6f (KF-20170) is now under further investigation as a drug for treating gastrointestinal disorder.