Acute phase response in toxicity studies. I. Survey of beagle dogs subjected to single-dose toxicity studies.

In the field of routine single-dose toxicity studies, we occasionally meet with transient leukocytosis associated with an increase in fibrinogen in beagle dogs within a few days after treatment with the test article. Only a little is known, however, about the toxicological significance of these changes. However, these changes were thought to belong to the category of "Acute Phase Response, APR," which has been known for a long time in connection with injury, trauma or infection. Aiming at proper understanding of these experiences, we surveyed 25 single-dose toxicity studies (7 intravenous bolus, 5 intravenous infusion, 12 oral and 1 subcutaneous treatment, hereafter referred to simply as i.v. bolus, i.v. infusion, oral and s.c.) in beagle dogs, provided with data from hematological examinations. We set the following criteria as a positive response in the present survey: increases of 50% or more in either or both WBC or fibrinogen compared to the predosing value, transiently from Day 1 to Day 3 of the study. Among 25 studies surveyed, about 1/2 of the studies exhibited increases of 50% or more in either or both fibrinogen or WBC counts compared to the predosing values showing dose-dependency transiently on Day 1 or Day 2. These changes were remarkable after intravenous application. Oral application produced similar effects, although the incidence and severity were low compared to the i.v. routes. Regarding blood chemical and hematological changes other than changes in fibrinogen and WBC counts, there were no essential differences between the groups of studies with and without the changes in fibrinogen and WBC counts. These changes were thought to be characteristic and to have occurred as incidents unrelated to other changes. The reported changes seen in single-dose toxicity studies may belong to the category of APR as the non-specific mechanism of living bodies as stated by Burns et al. (1996).

Antiulcer agents. III. Synthesis and antiulcer activity of N-[3-(3-piperidinomethylphenoxy)propyl]pentacyclo[4.2.0.0(2,5).0(3,8).0 (4,7)]-octane carboxamides and related compounds.

The synthesis and antiulcer activity of highly strained cage compounds such as pentacyclo[4.2.0.0(2,5).0(3,8).0(4,7)]-octane (cubane), pentacyclo[4.3.0.0(2,5).0(3,8).0(4,7)]nonane (homocubane) and pentacyclo[5.3.0.0(2,4).0(3,6).0(5,8)]decane are described. Of the compounds obtained, N-[3-(3-piperidinomethylphenoxy)propyl]-4-piperidinocarbonylpen tacyclo [4.2.0.0(2,5).0(3,8).0(4,7)]octane carboxamide (26a) and N-[3'-(3'-piperidinomethylphenoxy)propyl]-1-bromo-9, 9-ethylenedioxypentacyclo[4.3.0.0(2,5).0(3,8).0(4,7)[nonane]-4- carboxamid e (26q) showed more potent antiulcer activity with very good cytoprotective ability in the HCl.ethanol-treated rat model. Compounds 26a and 26q exhibited H2-receptor antagonist potency (in vitro) comparable to that of ranitidine, but did not inhibit histamine-stimulated acid secretion (in vivo) in the gastric fistula rat model, when orally administered in the dose range at which antiulcer and cytoprotective activities were seen. The structure-activity relationships are discussed.

Inhibition of bovine leukocyte thioltransferase by anti-inflammatory drugs and anti-histaminic drugs.

Thioltransferase was partially purified from bovine leukocyte using sonication, heat treatment at pH 5.1, Sephadex G-50 gel filtration and isoelectric focusing techniques. Isoelectric point (pI) of 8.3 for leukocyte thioltransferase was quite different from pI of 6.5 for erythrocyte enzyme. Bovine leukocyte thioltransferase was employed in a study on the influence of 12 anti-inflammatory drugs and 7 anti-histaminic drugs. Piroxicam which is a well-known anti-inflammatory drug demonstrated the most powerful inhibitory effect on enzyme activity. Inhibition of piroxicam was noncompetitive, and the Ki value measured 55.0 microM in the experiment involving bovine leukocyte enzyme. Tranilast which is a typical anti-histaminic drug most strongly inhibited the enzyme activity and the Ki value of the medicine was 20.3 microM (noncompetitive). Bovine liver and erythrocyte thioltransferases also were effectively inhibited by both medicines similar to leukocyte enzyme.

Purification and characterization of carbonyl reductases from bovine liver cytosol and microsome. The cytosolic enzyme has a novel 3 alpha/17 beta-hydroxysteroid dehydrogenase activity.

1. Carbonyl reductase, which is distributed in both cytosolic and microsomal fractions in bovine liver, were purified to homogeneity on 12.5% sodium dodecylsulfate-polyacrylamide gel electrophoresis and shown to have molecular weights of 32 kDa and 68 kDa, respectively. 2. Both carbonyl reductases can catalyze the reduction of many carbonyl compounds including ketone, quinones and aldehyde with relatively low Km values. 3. From the absorption spectrum result, microsomal carbonyl reductase closely resembles cytochrome P-450 reductase. 4. Cytosolic carbonyl reductase is a novel enzyme which can act on both testosterone and androsterone at low concentration.

[Injuries of the sigmoid colon following radiation therapy of carcinoma of the uterine cervix--a method of estimating the radiation dose to the sigmoid colon].

Grade 2 or 3 injuries of the sigmoid colon were observed in 4 out of 42 patients with carcinoma of the uterine cervix who were treated by radiation therapy. The irradiation was planned as the combination of the external irradiation (whole pelvic 30 Gy and 20 Gy with central shielding by 25 fractions, 5 weeks) and the intracavitary irradiation (RALS, 19 Gy at point A by 3 fractions). To analyze the causes of the radiation sigmoiditis, we have investigated the following factors: age, dose at point A, dose at point C, grade of tandem dislocation, uterine angle, obesity score, evidence of previous surgery to the pelvic cavity and hypertension. The dose at point C and the grade of tandem dislocation were determined from the confirming X-Ps at RALS therapy and external irradiation. The superimposition of these films was performed with corrections for the angle between the projection direction of the X-Ps and the vertical magnification factor of the central shielding area. Point C was defined as a point 2 cm anterior to the intersection of the tandem axis and a curvilinear line 1 cm outside from the margin of central shield on the X-Ps. Grades of tandem disclocation were decided as the number of tandem tips which were outside of the central shielding area on X-Ps. As the results, the doses at point C showed very high statistical significance (p less than 0.001) with the evidence of radiation sigmoiditis. All the cases with radiation sigmoiditis were received over 1290cGy at point C. Age had also some significance (p less than 0.05) with radiation sigmoiditis.(ABSTRACT TRUNCATED AT 250 WORDS)

[Radiotherapy of uterine cervix carcinoma].

Seventy-eight patients with uterine cervix cancer were treated radically with standardized radiation therapy at Teikyo University Hospital in Tokyo from January 1979 to December 1985. The age of these patients ranged from 32 to 88 years old (average age 66.0). The pathology and the stage of them were 71 cases of squamous cell carcinoma (1 in stage I, 11 in stage II, 58 in stage III and 1 in stage IV) and 7 cases of adenocarcinoma (6 in stage III and 1 in stage IV). The cumulative survival rates for 5 years by Kaplan-Meier method were 71.5% for squamous cell carcinoma stage II, 47.4% for squamous cell carcinoma stage III and 0% for adenocarcinoma stage III. Radiation injury was studied by the grading system of Kottmeier-NIRS in Japan. The incidence of the injuries for grade 2 and 3 was 12.8% (10/78), and the items of those were rectal injury 5.1% (4/78) and sigmoidal colonic injury 7.7% (6/78). The results of survival rate were fair for the squamous cell carcinoma compared with the other reports but poor for adenocarcinoma stage III. Concerning the therapy for advanced adenocarcinoma of the uterine cervix, special consideration should be given for irradiation doses or infusion chemotherapy.

[The effects of angiographic contrast media on the aggregation and morphology of red cells in vitro].

The effects of four angiographic contrast media on the aggregation and morphology of human red cells in vitro, using microscopic observations were studied. The media included an ionic contrast medium, sodium meglumine amidotrizoate (amidotrizoate); non-ionic low-osmolal contrast media, iopamidol and iohexol; and an ionic low-osmolal contrast medium, sodium meglumine ioxaglate (ioxaglate). Strong, large aggregates formed in the control blood, without media, where aggregation of red cells was inhibited by contrast media mixed with the blood in a ratio of 2:1. Almost no aggregates were observed for amidotrizoate, an ionic contrast medium, while there were a few rouleaux formed in the presence of ioxaglate. Nearly all of the red cells aggregated in the presence of iopamidol and iohexol; iohexol produced the greater aggregation of the two. Besides rouleaux, irregular aggregates were formed with iohexol. When the contrast media were mixed with blood in a ratio of 1:2, their inhibitory effects on aggregation declined. These results clearly indicate that contrast media inhibit the in vitro aggregation of red cells, and ionic-contrast media produced more potent inhibitory effects than non-ionic media. With added NaCl and meglumine, iohexol did not induce red cell aggregation. This suggests that ionic-contrast media have greater inhibitory effects on aggregation than non-ionic media, a result of their ionic properties. Red cells were morphologically quite normal in the presence of ioxaglate, where most red cells were crenated in the presence of iopamidol and iohexol, and shrank in the presence of amidotrizoate. In the presence of iopamidol and iohexol with the osmolality adjusted to that of a saline solution, both normal red cells and crenation were observed. This suggests that non-ionic contrast media may directly effect morphological changes in red blood cells. These results revealed that ioxaglate, an ionic contrast medium, was the best in vitro medium, to prevent aggregation of red cells and crenation deformity of erythrocytes.

The effects of the administration routes on the biliary excretion of antibiotics.

The effects of the administration routes on the biliary excretion of gentamicin and cefazolin were investigated in male white rabbits. The administration routes studied were intravenous, intramuscular injection and injection into portal vein. In both antibiotics, total excretion into bile was the highest when drug was administered by the injection into portal vein. In case of gentamicin, the bile level above MIC (against P.aeruginosa) could be obtained only by the administration into portal vein. These results indicate that the administration into portal vein is useful clinically, for the drug delivery to biliary tract, especially in drugs like aminoglycoside antibiotics which have an extremely low rate of transfer to the bile.