Bovine serum was subjected to ammonium sulfate fractionation, and subsequently chromatographed on DEAE-cellulose column to obtain a globulin protein having a weak auti-ulcerogenic activity, Fr. II-A. Agar immunoelectrophoresis and immunodiffusion using rabbit antiserum against bovine IgG revealed that Fr. II-A from bovine serum shared antigenic determinant with bovine IgG. Furthermore, Fr. II-A showed a single band on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. By reducing and alkylating the disulfide bonds by 2-mercaptoethanol and iodoacetic acid, Fr. II-A was separated into two subfragments in the same manner as in the case of immunoglobulin to produce Fr. L (molecular weight ; 24000) and Fr. H (molecular weight ; 50000). Fr. L showed anti-ulcerogenic activity up to about ten times greater than that of original Fr. II-A, but in the case of Fr. H, the activity increased approximately several times. Fr. L showed significant activity in preventing ulcer formation and gastric juice secretion in pylorus-ligated rats, and was also effective in reducing index of the phenylbutazoneinduced ulcer.

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Relationship between contraction and cyclic AMP levels induced by BaCl₂ was examined in the longitudinal smooth muscle isolated from guinea pig ileum. BaCl₂ 3×10^-3M caused a fast initial contraction, often followed by a gradual decrease of the contractile state. There was an increase in the tissue cyclic AMP 7 min or 14 min after the application of Ba. A phosphodiesterase activator imidazole reinforced the later phase of contraction by Ba and inhibited the increase in cyclic AMP. These results indicate that there is still a positive correlation between relaxation and increase in cyclic AMP and that an inhibitory action mediated by cyclic AMP is veiled behind the Ba contraction. Furthermore, these results may be interpreted by assuming that strong Ba contraction operates a feedback mechanism and that the feedback mechanism is associated with cyclic AMP increase. Indomethacin, an inhibitor of prostaglandins synthesis, little influenced the Ba-induced increase in cyclic AMP and rather inhibited the Ba contraction. Propranolol, a beta-adrenergic blocking agent, failed to exert influence on the Ba contraction. Based on these facts, it is suggested that the increase in cyclic AMP is not mediated by prostaglandins or catecholamines.

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Esterases of intestinal mucosa and liver from human and rat were used to study the biotransformation of midecamycin derivatives. In the in vitro experiment with rat esterases, the 4"-acyl derivatives were more easily hydrolyzed than the 9-acyl derivatives. Among the 9-acyl esters, the highest hydrolytic activity was observed with butyrate. In the in vivo experiment, when the rats were administered with the derivatives of 4"-depropionylmidecamycin (Ml) orally, comparatively more 9-acyl metabolites were excreted in the urine, but the amount of the 4"-acyl metabolites was very small. In the in vitro experiment with human esterases, the 9-acyl esters were hydrolyzed more easily than the 4"-acyl esters. Among the 9-acyl esters of Ml, the highest hydrolytic activity was observed with butyrate. When the 9-acyl esters were administered to humans, the n-butyl ester was hydrolyzed faster than the acetyl ester. When the 9, 4"-diacetyl ester was administered to humans, comparatively more 4"-acyl metabolites were excreted in the urine. These results suggest that the experiment with the use of these esterases is useful to estimate the biotransformation of midecamycin derivatives.

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Gelatinase, elastase and cathepsin G isolated from the granule extract of rat polymorphonuclear leukocytes (PMNs) had similar properties to the enzymes of human PMNs already reported. Effect of proteinase inhibitors on these neutral proteinases isolated from rat PMNs was studied. ε-Amino-n-caproic acid n-hexyl ester, a proteinase inhibitor having anti-inflammatory activity, inhibited cathepsin G, whereas elastase was activated by the inhibitor. On the other hand, leupeptin, L-1-tosylamide-2-phenylethyl chloromethyl ketone and N-a-p-tosyl-L-lysine chloromethyl ketone, which had been reported as antiinflammatory inhibitors, had no inhibitory effect on these neutral proteinases. These results suggest that proteinase inhibitors reported as anti-inflammatory agents exert their anti-inflammatory actions not by direct inhibition of the neutral proteinases released from PMNs, but by other effects such as suppression of the infiltration of PMNs into inflammatory locus.

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The effects of cortisol and testosterone on calcium metabolism in the hepatic bile system were investigated in thyroparathyroidectomized rats. A single subcutaneous administration of cortisol (0.1 mg/100 g) or testosterone (0.1 mg/100 g) caused a significant fall in serum calcium and a corresponding increase in liver calcium. On the other hand, the excretion of calcium into the bile after a single intraperitoneal injection of calcium chloride (4.0 mg Ca/100 g) was markedly increased by the administration of cortisol (0.1 and 1.0 mg/100 g) or testosterone (0.1 and 1.0mg/100 g). However, the bile calcium excretion was not significantly altered by the administration of thyroxine (0.1 and 1.0 mg/100 g) or vitamin D₃ (20 and 100 μg/100 g). The present results suggest that the hypocalcemic effect of cortisol or testosterone is partly based on the stimulation of calcium excretion into the bile by the hormones.

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Influence of aggregation on the action of methamphetamine in general behavior was investigated in mice. The action of increasing the amount of movement by methamphetamine was significantly increased by aggregation compared to isolation of mice after administration of 5, 15 and 30 mg/kg, i.p., suggesting the group toxicity of methamphetamine.

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4-¹⁴C-Estradiol and its 17β-sulfate were incubated with rat liver microsomes under NADPH-generating system. Estradiol in liver microsomes from male and female rats was metabolized to multiple kinds of oxidized products including estrone, 2-hydroxyestrone, 2-hydroxyestradiol, and other minor steroids. Incubation of estradiol 17β-sulfate was carried out by the same condition, and it was shown that the metabolic pattern between male and female rats was different. By incubation of estradiol 17β-sulfate with male rat liver microsomes, 2-hydroxyestradiol 17β-sulfate was obtained as the sole product (6%). The hydroxylation was shown to occur without cleavage of the conjugate group. No such regulating effect by conjugate group on 2-hydroxylation of estradiol 17β-sulfate was observed in liver microsomes from female rats. The amount of 2-hydroxyestradiol 17β-sulfate formed was only 1%, and other metabolites which were thought to be monohydroxylated estradiols were produced as the major products. The 2-hydroxylated metabolite of estradiol 17β-sulfate was confirmed by its isolation as a stable form of derivative by the following way. The incubation mixture of massive amount of estradiol 17β-sulfate was extracted with n-butanol. Methylation of the extract with diazomethane, followed by acid-catalized hydrolysis, acetylation, and finally separation by preparative thin-layer chromatography, gave a crystalline material, the spectral properties of which were completely identical with those of the synthetic specimen, 2, 3-dimethoxy-1, 3, 5 (10)-estratrien-17β-y1 acetate.

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Intravenous injection of antigenic dose of sheep erythrocytes (SRBC)^* into mice caused a 150-200% increase in plasma cyclic GMP level within 5 min which continued for 60 min thereafter. Immunization by soluble antigens such as dextran sulfate and bovine serum albumin also elevated cyclic GMP level in plasma. The plasma cyclic GMP increased by antigen stimulation might be derived from immunocompetent cells. This assumption was supported by several lines of evidence as follows : (1) rat erythrocytes which are less antigenic for mice caused a relatively low response in terms of increase in cyclic GMP compared with SRBC, (2) this cyclic GMP response was abrogated in animals under immunosuppressive states where X-irradiated, azathioprine-treated or tumor-bearing mice were used. Mice pretreated with agents which block autonomic nervous system functions develop normal plasma cyclic GMP responses upon SRBC injection. Our observation in this report and many in vitro studies by other investigators suggested that plasma cyclic GMP elevated by antigen stimulation may be mainly derived from lymphoid cells as a consequence of the triggering of immune response.

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The effects of dilazep on blood fibrinolytic system were studied in ex vivo experiment. Fibrinolytic activity in plasma was determined by the fibrin plate method at designed times after the administration of dilazep in guinea pigs, rabbits and mongrel dogs. In guinea pigs, fibrinolytic activity of euglobulin (plasminogen activator level, 1.10±0.08 CU/ml in control animals) rose to 1.53±0.24, 1.92±0.13 and 2.35±0.15 CU/ml 2 h after the p.o. administration of 30, 100 and 300 mg/kg of dilazep, respectively. Plasminogen levels in euglobulin were unchanged, however, plasmin inhibitory activities were lowered following the increase of plasminogen activator levels. About 2-fold increase of plasminogen activator level was also observed in rabbits 60 min after the p.o. administration of 300 mg/kg dilazep and returned to the initial level (0.054 CU/ml) within 3 h after the administration. A rapid increase of plasminogen activator level was observed in a case of the i.v. administration of dilazep into rabbits and mongrel dogs. In in vitro experiment, dilazep added to whole blood or plasma of guinea pigs ranging 0.1 to 1000 μg/ml did not reveal any effects on plasminogen activator level or plasmin inhibitory activity in plasma.

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The influence of drugs on the release of plasminogen activator was studied in the isolated perfused pig ear. The pig ear was perfused with oxygenated Tyrode's solution pH 7.4, at 37°C via main artery and perfusate from the veins was collected at 2 min intervals. The drug was injected into a rubber tube connected in the front of arteria cannula, and the fibrinolytic activity of perfusate collected was measured by the fibrin plate method using plasminogen-containing bovine fibrinogen. Under the test condition, dilazep enhanced plasminogen activator release (PA release) in a dose-dependent fashion ranging 10 to 100 μg, however, it did not affect the perfusion pressure. Hypotensors, namely acetylcholine (0.1-3μg), bradykinin (0.1-3μg) and histamine (0.1-3μg) enhanced also the PA release in a dose-dependent fashion with no effects on perfusion pressure. Vasoconstricting drugs, namely phenylephrine (0.3-10 μg), norepinephrine (0.1-3 μg) and serotonin (0.1-3μg) exerted hypertensive effects on perfusion pressure in a dose-dependent fashion, however, it did not cause the PA release.

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The mean in vitro dissolution times (in vitro MDT) of ampicillin products were evaluated from the dissolution curves in distilled water using the USP-XX dissolution apparatus, and were compared with the mean in vivo dissolution times (in vivo MDT) reported previously. In both in vivo and in vitro cases, MDT of the trihydrate capsule was greater than those of the anhydrate capsules. A good linear correlation was observed between in vivo and in vitro MDT (r=0.999, p<0.001).

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