Expression of organic anion transporting polypeptide E (OATP-E) in human placenta.

PURPOSE: To examine the expression of multifunctional Na(+)-independent organic anion transporting polypeptide, termed OATP-E, involved in the transport of thyroid hormone in human placenta. METHODS: Western blot analysis was performed using a specific antibody against OATP-E in human placenta to confirm the expression of OATP-E at the protein level. Immunohistochemistry was also performed using the specific antibody against OATP-E on frozen sections of human placenta. RESULTS: By Western blot analysis, a single band for OATP-E was observed in human placenta. Immunohistochemistry revealed that OATP-E was predominantly expressed at the apical surface of the syncytiotrophoblasts in placenta. CONCLUSION: These results reveal that OATP-E is expressed in human placenta, suggesting a functional role for the transplacental transfer of thyroid hormone.

[Epidemiological survey for hemolytic streptococci isolated from children in Tokyo].

During the 20-year period between 1979-1998, a total of 4,176 strains of hemolytic streptococci have been isolated from 20,118 healthy primary school children and little children in Tokyo Metropolitan (Tokubetsuku, Tama and Tosho). Culture of throat swabs every November and the following February during the 20-year period were made and serological grouping and typing for isolates were done by T agglutination method. The results were as follows. 1) Serological group of hemolytic streptococci isolated from children were 3,188 strains (76.3%) for isolates of group A out of total strains of 4,176, 569 strains (13.6%) for isolates group B, 63 strains (1.5%) for isolates of group C and 356 strains (8.5%) for isolates of group G. 2) The most dominant was T12 during 1979-1998, and other relatively frequent serotypes were T28, T1, T4, T6 in that order. These ranks of and the main epidemic serotypes showed a similar trend in the 3 areas. 3) The isolation rates of group A streptococci were 15.9% in Tokubetsuku, 17.1% in Tama and 14.9% in Tosho. The average of 3 areas were 15.8%. 4) The epidemic cases seemed to be caused by group A streptococci were 20 cases, their isolated serotype were 7 cases by T28, 5 cases by T12, 4 cases by T6, 2 cases by T4, each 1 case by T1 and T25. 5) A total of 2,927 strains of group A streptococci were examined for drug sensitivity. All strains were sensitive to beta-lactam group of antibiotics (benzylpenicillin and cephaloridine). Resistant (MIC > or = 25 micrograms/ml) to TC, CP and EM etc. were 740 strains (25.3%) in this study. The incidence of resistant strains were to TC 493 strains (66.6%) out of 740 strains, 81 strains (10.9%) for TC.CP, 72 strains (9.7%) for EM and 66 strains (8.9%) for TC.CP.EM.OL.LCM. TC resistant strains have not varied much through the whole period, but CP and EM resistant strains were very variable by year. Many resistant strains to TC were T4, to EM and multiple drug resistant were T12. 6) The rates of isolates of the same type of group A streptococci in school child individual during for the tests taken twice a year were 12.3%, indicating group A streptococci, according to the duration of the carrier state, seems to be a short period.

LST-2, a human liver-specific organic anion transporter, determines methotrexate sensitivity in gastrointestinal cancers.

BACKGROUND & AIMS: One approach to the development of targeted cancer chemotherapy exploits increased uptake of the agent into neoplastic cells. In this scenario, higher concentrations of the agent in cancer cells are responsible for differential killing, whereas the low concentration in normal human cells decreases side effects. The aim of this study was to isolate an organic anion transporter that is weak in normal cells, but abundantly expressed in cancer cells, to deliver the anticancer drugs to the cells. METHODS: A human liver complementary DNA (cDNA) library was screened with liver-specific transporter (LST)-1 cDNA as a probe. Northern blot analyses were performed using the isolated cDNA (termed LST-2). An LST-2-specific antibody was raised, and immunohistochemical analyses including immunoelectron microscopy were performed. Xenopus oocyte expression system was used for functional analysis. We also established a permanent cell line that consistently expresses LST-2 to examine the relationship between methotrexate uptake and sensitivity. RESULTS: The isolated cDNA, LST-2, has 79.7% of overall homology with human LST-1. LST-2 exclusively expressed in the liver under normal conditions and its immunoreactivity was highest at the basolateral membrane of the hepatocytes around the central vein. Although its weak expression in the liver, LST-2 is abundantly expressed in the gastric, colon, and pancreatic cancers. On the other hand, the LST-1 was only detected in a hepatic cell line. LST-2 transports methotrexate in a saturable and dose-dependent manner. Furthermore, introduction of the LST-2 gene into mammalian cells potentiates sensitivity to methotrexate. CONCLUSIONS: LST-2 is one of the prime candidate molecules for determining methotrexate sensitivity and may be a good target to deliver anticancer drugs to the gastrointestinal cancers.

Identification of thyroid hormone transporters in humans: different molecules are involved in a tissue-specific manner.

We have recently identified that rat organic anion transporters, polypeptide2 (oatp2) and oatp3, both of which transport thyroid hormones. However, in humans the molecular organization of the organic anion transporters has diverged, and the responsible molecule for thyroid hormone transport has not been clarified, except for human liver-specific transporter (LST-1) identified by us. In this study we isolated and characterized a novel human organic anion transporter, OATP-E from human brain. The isolated complementary DNA encodes a polypeptide of 722 amino acids with 12 transmembrane domains. A rat counterpart, oatp-E, was also identified. Homology analysis and the phylogenetic tree analysis revealed that OATP-E/oatp-E is a subfamily of the organic anion transporter. Human OATP-E transported 3,3',5-triiodo-L-thyronine (K(m), 0.9 microM), thyronine, and rT(3) in a Na(+)-independent manner. Although the clone was isolated from the brain, OATP-E messenger RNA was abundantly expressed in various peripheral tissues. The rat counterpart, oatp-E, also transported 3,3',5-triiodo-L-thyronine. In addition, in this study we revealed that human OATP, which is exclusively expressed in the brain, transported 3,3',5-triiodo-L-thyronine (K(m), 6.5 microM), T(4) (K(m), 8.0 microM), and rT(3). These data suggest that in humans, several different molecules are involved in transporting thyroid hormone: OATP in the brain, LST-1 in the liver, and OATP-E in peripheral tissues.

Molecular identification of a rat novel organic anion transporter moat1, which transports prostaglandin D(2), leukotriene C(4), and taurocholate.

We have isolated a rat novel multispecific organic anion transporter, moat1. The isolated clones were originated by alternative splicing of the moat1 mRNA. The nucleotide sequences predict a protein of 682 amino acids with moderate sequence similarity to LST-1, the oatp family, and the prostaglandin transporter. Northern blot analysis of rat moat1 identified a predominant transcript of 4.4 kilonucleotides in all tissues. Northern blot and in situ hybridization analyses of rat brain further indicated that moat1 mRNA is widely distributed in neuronal cells of the central nervous system, especially in the hippocampus and cerebellum. moat1 transports prostaglandin D(2) (K(m); 35.5 nM), leukotriene C(4) (K(m); 3.2 microM) and taurocholate (K(m); 17.6 microM) in a sodium-independent manner. moat1 also transports prostaglandin E(1), E(2), thromboxane B(2), and iloprost but not dehydroepiandrosterone sulfate and digoxin, of which the substrate specificity is similar, but definitively different from those of any other organic anion transporters.

Effects of the V(2)-receptor antagonist OPC-41061 and the loop diuretic furosemide alone and in combination in rats.

This study was conducted to characterize the diuretic effect of OPC-41061, a nonpeptide vasopressin V(2)-receptor antagonist, and furosemide by administering each alone and in combination in conscious male rats. OPC-41061 at 1 and 10 mg/kg and furosemide at 10 and 100 mg/kg dose-dependently increased urine volume to the same extent. The high dose of OPC-41061 (10 mg/kg) markedly elevated electrolyte-free water clearance (E-CH(2)o) to a positive value. In contrast to OPC-41061, furosemide elevated only electrolyte clearance but not E-CH(2)o. The differences in diuretic profile reflected the changes in serum sodium and hormone levels. OPC-41061 dose-dependently elevated serum sodium concentration, but furosemide tended to decrease it. The high dose of furosemide (100 mg/kg) significantly elevated serum renin activity and aldosterone concentration, indicating that furosemide activated the renin-angiotensin-aldosterone system (RAA-system). On the other hand, OPC-41061 did not affect these parameters. When OPC-41061 was administered concomitantly with furosemide, OPC-41061 significantly increased urine volume and E-CH(2)o, and decreased urinary osmolality compared with furosemide alone. OPC-41061 dose-dependently elevated serum osmolality and sodium concentration even when administered in combination with the high dose of furosemide. These results suggest that OPC-41061 produces aquaresis leading to increased serum sodium without affecting the RAA-system. On the other hand, furosemide produced natriuresis, leading to decreased serum sodium level and activation of the RAA-system. It was also demonstrated that OPC-41061 produced an additive diuretic effect and elevated serum sodium level in the presence of furosemide.

[Effect of complement on the production of tumor necrosis factor alpha by human leukocytes with culture supernatants of clinical isolates of Staphylococcus aureus].

Induction of tumor necrosis factor alpha (TNF-alpha) of human leukocytes by overnight culture supernatants of Staphylococcus aureus (S. aureus) (CS) (4 strains) and then the effect of the complement on production of TNF alpha by the treated cells were examined. One-tenth diluted solution of CS was added to Heparinized blood and was incubated for 6 hrs at 37 C in a 5% CO-air humidified incubator. TNF alpha level in the plasma was measured by ELISA. The level in the plasma was different in different CS. The induction of TNF alpha was not found in EDTA-added blood but was found in the EGTA-added them treated with CS, though different levels were showed. Western blotting assay of these plasma samples without EDTA-added blood found the presence of both C5a and C3a. Neither C5a and C3a was found in Heparinized blood treated with a relative concentration to TSST-1, enterotoxin C and alpha (alpha) homolysin contained in CS. Mixture of these toxins induced only 1/6.5 of the TNF alpha amount obtained with the CS. Human leukocytes isolated by Mono.-Poly. resolving solution were cultured with CS in RPMI 1640 medium with or without 10% fresh or heated serum. Production of TNF alpha by the isolated leukocytes was not found under the condition of serum free. But it was found in the presence of fresh serum and also in the presence of heated serum. The addition of CS to RPMI 1640 containing heated serum did not occur naturally in the production of C5a. These results suggest that there is a difference in TNF alpha inductivility to human leukocytes in each strain and that other bacterial components without TSST-1, SEC and alpha-hemolysin are more important to induce TNF alpha. Serum may be necessary to produce TNF alpha by CS-treated cells as a source of factor(s) to interact with bacterial components rather than a source of complement.

7-Chloro-5-hydroxy-1-[2-methyl-4-(2-methylbenzoyl-amino)benzoyl ]-2,3,4,5-tetrahydro-1H-1-benzazepine (OPC-41061): a potent, orally active nonpeptide arginine vasopressin V2 receptor antagonist.

We previously reported a series of benzazepine derivatives as orally active nonpeptide arginine vasopressin (AVP) V2 receptor antagonists. After the lead structure OPC-31260 was structurally evaluated and optimized, the introduction of the 7-Cl moiety on the benzazepine and 2-CH3 on the aminobenzoyl moiety enhanced its oral activity. The new AVP-V2 selective antagonist OPC-41061 was determined to be a potent and orally active agent.

[Soluble proteins from Staphylococcus aureus can change expression of CD11b and CD62L, but not H2O2 production by human blood granulocytes].

We examined the production of CD11b, CD62L and H2O2 by human peripheral blood granulocytes after treatment with soluble proteins prepared from five different pressure-disrupted strains of Staphylococcus aureus (SaSP) by flow cytometory. Peripheral blood was treated with final SaSP concentrations of 0.05, 0.5 and 5.0 micrograms for 20 min at 37 degrees C. The ratio of CD11b positive granulocytes did not increase at concentrations from 0.05 to 5.0 micrograms, but fluorescence intensity showed about two and three-fold increase, respectively, at concentrations of 0.5 and 5.0 micrograms, in comparison with that of control cells. The ratio CD62L positive cells decreased as follows: 0.5 microgram, 53.8% and 5.0 micrograms, 19.0%, respectively, whereas the control value was 79.8%. Fluorescence intensity also decreased as follows: 0.5 microgram, 10.4 and 5.0 micrograms, 9.2, respectively, whereas the control value was 46.8. Slight induction of H2O2 was found at 5.0 micrograms concentration only. In addition, SaSP treatment granulocytes that stimulated with PMA (1 ng) increased H2O2 production. Thus, SaSP has no beneficial effect against H2O2 production by granulocytes. All of the SaSP preparations indicated similar results for the production of CD11b, CD62L and H2O2 by granulocytes. SaSP effects activation of granulocytes, and the activation may occur independently of protein kinase C.

OPC-41061, a highly potent human vasopressin V2-receptor antagonist: pharmacological profile and aquaretic effect by single and multiple oral dosing in rats.

The pharmacological profile and the acute and chronic aquaretic effects of OPC-41061, a novel nonpeptide human arginine vasopressin (AVP) V2-receptor antagonist, were respectively characterized in HeLa cells expressing cloned human AVP receptors and in conscious male rats. OPC-41061 antagonized [3H]-AVP binding to human V2-receptors (Ki = 0.43 +/- 0.06 nM) more potently than AVP (Ki = 0. 78 +/- 0.08 nM) or OPC-31260 (Ki = 9.42 +/- 0.90 nM). OPC-41061 also inhibited [3H]-AVP binding to human V1a-receptors (Ki = 12.3 +/- 0.8 nM) but not to human V1b-receptors, indicating that OPC-41061 was 29 times more selective for V2-receptors than for V1a-receptors. OPC-41061 inhibited cAMP production induced by AVP with no intrinsic agonist activity. In rats, OPC-41061 inhibited [3H]-AVP binding to V1a-receptors (Ki = 325 +/- 41 nM) and V2-receptors (Ki = 1.33 +/- 0. 30 nM), showing higher receptor selectivity (V1a/V2 = 244) than with human receptors. A single oral administration of OPC-41061 in rats clearly produced dose-dependent aquaresis. In treatment by multiple OPC-41061 dosing for 28 days at 1 and 10 mg/kg p.o. in rats, significant aquaretic effects were seen throughout the study period. As the result of aquaresis, hemoconcentration was seen at 4 hr postdosing although, no differences were seen in serum osmolality, sodium, creatinine and urea nitrogen concentrations at 24 hr postdosing. Furthermore, there was no difference in serum AVP concentration, pituitary AVP content or the number and affinity of AVP receptors in the kidney and liver at trough throughout the study period. These results demonstrate that OPC-41061 is a highly potent human AVP V2-receptor antagonist and produces clear aquaresis after single and multiple dosing, suggesting the usefulness in the treatment of various water retaining states.

[Difference of host response in identical toxin-produced Staphylococcus aureus-injected mice].

Biological activities of two strains of Staphylococcus aureus (S. aureus), KU-1-06-37 and KU-1-12-44, which produce enterotoxin type C, TSST-1 and alpha-toxin were examined using Std:ddY strain mice. These two strains were found to be different in lethality, ratio of weight loss, induction of leukopenia, adhesion to surrounding organs and clearance period of bacterial cells from the liver, kidney and spleen within 24 hrs after intraperitoneal injection in the mice. All of them were weak or fast in KU-1-12-44 injected mice. Serum amyloid A on all the KU-1-06-37 and KU-1-12-44 injected mice rose within 5 hr to 18 hr. However, this concentration of KU-1-12-44 injected mice was about 40% lower compared with that of KU-1-06-37 injected mice at 21 hr. On the other hand, ability of bacterial adhesion to established cell lines, Vero and HeLa cells, was tested in vitro. Percentage adhesion of KU-1-06-37 was high to both cells, but that of KU-1-12-44 was high to Vero cells and was low to HeLa cells. Adhesion of KU-1-06-37 to HeLa cells that were treated with lipoteichoic acid was about 40% inhibition compared with untreated cells, although that of KU-1-12-44 to them was inhibited only 9%. As the results, identical toxin-produced KU-1-06-37 and KU-1-12-44 showed different biological activities in vivo and in vitro. Not only toxin production but also adhesion to cells or organs in mice may contribute to S. aureus virulence to the host.

[Inducibility of tumor necrosis factor alpha to peritoneal macrophages and lethality to mice of clinical isolates of Staphylococcus aureus].

When murine resident peritoneal macrophages were treated in vitro with overnight culture supernatants of clinical isolates of S. aureus (21 strains)(S. aureus-CS) for 4 hrs at 37 degrees C in a 5% CO2-air humidified incubator, TNF alpha induction from the treated cells was observed in 20/21 strains. The amount of TNF alpha dispersed from 88.4 to 1726.5 pg/ml but more or less amount of TNF alpha did not relate with the presence of TSST-1 and enterotoxin production of S. aureus. Recombinant protein A induced TNF alpha production by macrophages dose-dependently, but the relationship between the amount of protein A in each S. aureus CS and that of TNF alpha induced from macrophages treated with them were not satisfactory (r = 0.69). When each strain was injected (X10(9) bacterial cells) interperitonealy to mice, 13/21 strains indicated lethal activity. Relation between TNF alpha inducibility in vitro and lethal activity, however, was not found (r = 0.4). These results suggest that TNF alpha inducibility is the basic biological activity of S. aureus, although there is a difference in the induction amount and in component(s) to induce TNF alpha on each strain, and then because the strain is strong in TNF alpha inducibility, it does not necessarily follow that pathogenicity is strong.

[Effects of Staphylococcus aureus protein A on the production of primary host defense factors against S. aureus infection].

In vitro examination was carried out to investigate the effects of protein A of Staphylococcus aureus (S. aureus) on the production of fibronectin (Fn) and the third component of complement (C3) by macrophages and that of CD11b, CD49e and H2O2 by granulocytes. Fn production of cultured murine peritoneal macrophages (M phi ) increased significantly (P < 0.01) by treatment for 4 hr at 37 degrees C with recombinant PA (rPA) and the supernatant of overnight culture of S. aureus Cowan I strain (CoCS) (p < 0.01), not that of Wood 46 strain (WoCS), in comparison with that of control. The activity of rPA was inhibited strongly in the presence of CYH (1.0 microgram/ml). The production of C3 by cultured M phi did not increased by treatment for 4 hr at 37 degrees C with rPA, CoCS and WoCS. In these cells treated with CoCS and WoCS, however, the production increased by cultivation in serum free medium for a further 20 hr at 37 degrees C after the treatment. But increase was not found in rPA treated cells. On the other hand, the production of Mac-1, VLA-5 and H2O2 by granulocytes did not increase by treatment with rPA and CoCS. These results show that rPA and PA in CoCS are major components which stimulates Fn synthesis and secretion by cultured M phi some component(s) contained in the supernatant of overnight culture of S. aureus increased the production C3 by cultured M phi , and the supernatant may not contain stimulators to induce production of CD49e, CD11b and H2O2. Increase of Fn production of M phi by PA stimulation may play an important role in the primary host defense against S. aureus infection.

[Isolation frequency and biological characteristics of the multiple-antibiotic resistant Pseudomonas aeruginosa isolated from clinical specimens].

Isolation frequency of multiple-antibiotic resistant Pseudomonas aeruginosa (MARPA) was 11.9% (fifty six strains) of a total of four-hundred seventy-one strains of P. aeruginosa isolated from clinical specimens at the Kyorin University Hospital from October 1994 to December 1996. Eighteen strains of MARPA and thirteen strains of antibiotic sensitive P. aeruginosa (ASPA) isolated from clinical specimens in internal medicine ward A were determined O serotype, and characterized with production of pyocyanin, pyoverdin, hemolysin, elastase, and caseinase. Sixteen strains (88.9%) of MARPA were identified as serotype C. The ability to produce pyocyanin, hemolysin, elastase, and caseinase was not detected in all MARPA. One side, thirteen strains of ASPA showed various serotypes, i.e., B: 5 strains (38.4%), G: 4 strains (30.8%), C: 2 strains (15.4%), E: 1 strain (7.7%) and unknown type: 1 strain (7.7%), and the production of both hemolysin and pyoverdin was observed in 13 strains (100%), pyocyanin in 8 strains (61.5%), elastase and caseinase in 9 strains (69.2%) of ASPAs, which suggests that ASPAs do maintain the synthetic ability of pathogenic factors and pigments, but MARPAs do not. These results indicate that from epidemiological points of view the current strains of MARPA spread from one clone within the internal medicine ward A with nosocomial outbreak by serotype C.

OPC-21268 antagonizes arginine vasopressin-induced vasoconstrictor response in the spinally-anesthetized dog.

We studied the antagonistic actions of OPC-21268 (1-(1-[4-(3-acetylaminopropoxy)-benzoyl]-4-piperidyl)-3,4-dihydro- 2(1H)-quinolinone) on the arginine vasopressin (AVP)-induced vasoconstrictor response in the spinally-anesthetized dog. OPC-21268 at doses of 0.3, 1.0 and 3.0 mg/kg, i.v. produced a rightward parallel shift of the dose-response curves for AVP in a dose-dependent manner. The doses of OPC-21268 were similar to those that inhibited the AVP-induced vasoconstrictor response in the rat. This observation suggests that OPC-21268 acts as a V1-AVP-receptor antagonist in peripheral resistance vessels in dogs as well as in rats.

[Analysis of complement-mediated phagocytosis in systemic lupus erythematosus model mice, NZB/W F1].

We have attempted to examine the cause of lowering phagocytic activity of phagocytes on clearance of bacteria in SLE patients. The experiments were performed in SLE model mice, NZB/WF1 female mice (BWF1). The ratio of the cells which phagocytized fluorescent microspheres and possessed Mac-1 on both peripheral blood leukocytes (PBL) and peritoneal macrophages (PMo) in BWF1 female mice was shown decreasing by using the flow cytometric method before the present of typical symptoms of SLE (under 25 weeks-old), i.e., the appearance of ANA and IgG deposits in glomeruli. In a period of showing the symptoms of SLE (over 25 weeks-old), the ratio of Mac-1 positive cells on PBL increased, but phagocytic function of these cells did not increase. On the other hand, on PMo, they decreased together in that period. Moreover, the phagocytosis of PMo in 15, 25, 35 weeks-old mice against fluorescent microspheres showed a significant increase (1.7 times) by replacing opsonin with normal (ddY) murine serum instead of BWF1 murine serum, and the phagocytosis of PMo treated with anti-Mac-1 antibody (M1/70) showed a significant inhibition (46%) against that of untreated PMo. These results indicate that lowering phagocytic activity of phagocytes on primary defence against microbial infections in SLE patients is due possibly to decrease of C3 activity in the serum rather than a decrease of positive percentage and function of Mac-1 on these cells.

OPC-21268, a vasopressin V1 antagonist, produces hypotension in spontaneously hypertensive rats.

We studied the hypotensive effects of OPC-21268, an orally effective nonpeptide vasopressin V1 receptor antagonist, in spontaneously hypertensive rats (SHR) and stroke-prone SHR (SHRSP). OPC-21268 was given intravenously to conscious, freely moving SHR and SHRSP. We used young and aged animals to examine the contribution of vasopressin to the development and maintenance of hypertension in both types of rats. In SHR, hypertension was fully established at 38 weeks of age, and intravenous injection of OPC-21268 produced slight hypotensive effects at either 38 or 70 weeks of age. In SHRSP, hypertension developed at 25 weeks of age, and blood pressure was sustained at a high level (approximately 250 mm Hg systolic blood pressure) thereafter. Intravenous administration of OPC-21268 did not cause hypotensive effects in young rats at 15 weeks, but at 25 weeks a significant decrease in blood pressure was observed. Furthermore, in the malignant state of SHRSP (35 to 41 weeks), OPC-21268 significantly decreased mean blood pressure by 32.4 +/- 7.9 mm Hg (mean +/- SEM) at 3 mg/kg IV, and the decrease was dose dependent (0.3 to 3.0 mg/kg). Plasma vasopressin concentrations were increased in a more malignant phase of SHRSP at 45 weeks of age, whereas at other ages of SHRSP or in SHR, plasma vasopressin levels were not increased. These results suggest that vasopressin plays an important role through V1 receptors in the maintenance of hypertension, at least in the malignant phase of SHRSP, and OPC-21268 may be therapeutically useful in the treatment of some types of hypertension.

[Kinetics of the complement subcomponent C1q production of peritoneal macrophages in NZB/W F1 female mice].

To clarify the role of local production (exohepatic) of complement on primary host defense mechanisms against microbial infections in the host who decreased the amount of complement in serum, the kinetics of the complement production by the complement-producing cells in exohepatic tissue was examined by measuring the amount of C1q, subcomponent of the first complement component, in cultured supernatant of the monolayer of peritoneal macrophages (PM phi) collected from 5, 15, 35, and 48 weeks-old female mice of NZB/W F1 (B/W F1). The C1q production of PM phi in B/W F1 mice showed remarkable decrease at 15 weeks-old. After that, however, the C1q producibility of PM phi recovered gradually and, at mice 48 weeks old, the complement produced finally exceeded the amount observed in mice 5 weeks-old, contrariwise the C1q values in serum were significantly lowering in the same aged mice. The increased production of C1q of PM phi was observed in both 35 and 48 weeks-old mice, of which the value corresponded with increase of anti-nuclear antibody titer in serum and of the amount of protein in urine. On female mice of ddY (control), the amount of C1q production of PM phi in 5 weeks-old mice was two-fold higher than that of 5 weeks-old B/WF1 mice. But at 15 weeks, the production showed a 1/2 decrease in that of mice 5 weeks-old and the decreased values were kept to 35 weeks-old.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of a novel aquaretic agent, OPC-31260, as an orally effective, nonpeptide vasopressin V2 receptor antagonist.

1. OPC-31260, a benzazepine derivative, has been studied for its ability to antagonize the binding of arginine vasopressin (AVP) to receptors in rat liver (V1) and kidney (V2) plasma membranes, for antagonism of the antidiuretic action of AVP in alcohol-anaesthetized rats and for diuretic action in conscious normal rats. 2. OPC-31260 caused a competitive displacement of [3H]-AVP binding to both V1 and V2 receptors with IC50 values of 1.2 +/- 0.2 x 10(-6) M and 1.4 +/- 0.2 x 10(-8) M, respectively. 3. OPC-31260 at doses of 10 to 100 micrograms kg-1, i.v., inhibited the antidiuretic action of exogenously administered AVP in water-loaded, alcohol-anaesthetized rats in a dose-dependent manner. OPC-31260 did not exert an antidiuretic activity suggesting that it is not a partial V2 receptor agonist. 4. After oral administration at doses of 1 to 30 mg kg-1 in normal conscious rats, OPC-31260 dose-dependently increased urine flow and decreased urine osmolality. The diuretic action of OPC-31260 was characterized as aquaresis, the mode of diuretic action being different from previously known diuretic agents such as furosemide, hydrochlorothiazide and spironolactone. 5. The results indicate that OPC-31260 is a selective V2 receptor antagonist and behaves as an aquaretic agent. OPC-31260 will be a useful tool in studying the physiological role of AVP and in the treatment of various conditions characterized by water retention.