Measurement of plasma free and total growth hormone concentrations in patients with growth hormone antibodies developed in response to therapy.

Recombinant human growth hormone is more antigenic than pituitary preparations. Since GH antibodies interfere with radioimmunoassay of GH, we measured plasma free and total GH in patients with pituitary dwarfism with GH antibodies during treatment with a recombinant methionyl GH preparation. Plasma free GH was measured in the supernatant after polyethylene glycol precipitation. Total GH was measured after extracting plasma with acid-ethanol. In normal subjects, both free and total GH levels were similar to those measured in plasma by a direct conventional RIA. Peak free GH levels after administration of 4 IU methionyl GH to 3 normal subjects were 38.0, 30.7 and 13.2 micrograms/1, values similar to those measured in most of the patients with GH antibodies. All values were undetectable in one patient with a very high antibody titre. Total GH levels were similar to free GH levels in normal subjects. In patients with GH antibodies, total GH levels were high compared with their free levels, but similar to those assayed by conventional RIA. The patient with the highest antibody titre had total GH levels which were the lowest of those observed in the patients with GH antibodies in spite of having the highest GH levels measured by conventional RIA. The antibody in this particular case may have a high capacity and a high affinity. A relatively poor growth rate in this patient may be associated with the finding of undetectable free GH levels. Measurement of plasma free and total GH may be of value in examining GH dynamics and their relation to the clinical effectiveness of GH treatment in patients with GH antibodies.

The changes in plasma cortisol and urinary free cortisol by an overnight dexamethasone suppression test in patients with Cushing's disease.

We studied the suppressibility of cortisol secretion in 15 patients with Cushing's disease by measuring morning plasma cortisol level as well as the 24-hour urinary free corisol (UFC) excretion following single doses of increasing amounts of dexamethasone (ranging from 0.5 to 32 mg) given at 11 p.m. The mean plasma cortisol level in patients with Cushing's disease was twice as high as in normal subjects, whereas the mean UFC in these patients was 6 times as high. Plasma cortisol in seven patients were suppressed by less than 4 mg of dexamethasone (in 2 cases, less than 0.5 mg; in 3 cases, less than 2 mg; and in 2 cases less than 4 mg). In these cases, basal plasma cortisol and UFC were less than 25 micrograms/dl and 350 micrograms/day, respectively. Among the other eight patients, plasma cortisol was partially suppressed in 5 cases and not suppressed in 3 cases by high doses of dexamethasone (16-32 mg). In these cases the basal plasma cortisol and UFC were more than 25 micrograms/dl and 350 micrograms/day, respectively. There was a significant correlation between the basal plasma cortisol and UFC (r = 0.687, p less than 0.01). These data suggest that the suppression by increasing amounts of dexamethasone in most cases with Cushing's disease was related to the severity of hypercortisolism.

Hypercortisolism and the resistance to dexamethasone suppression during gestation.

Maternal adrenocortical function was studied by measuring plasma cortisol and urinary free cortisol during gestation. Changes in suppressibility of pituitary-adrenocortical function were determined by dexamethasone administration. Urinary free cortisol as well as plasma cortisol increased during the course of gestation. The suppressibility by dexamethasone became less effective as pregnancy advanced. These results suggest that pregnant women have pituitary-adrenocortical hyperfunction and tissue refractoriness to glucocorticoid which increases during the course of gestation.

Studies on pituitary-gonadal function in patients with Cushing's syndrome.

Basal levels of sex steroids, and the responses of LH and FSH to LH-RH were studied in twenty-five female patients with Cushing's syndrome (17 Cushing's disease and 8 adrenocortical adenoma). Only two patients had a regular menstrual cycle. Amenorrhea or oligomenorrhea had been of long duration in the other cases except for three postmenopausal patients. In patients with Cushing's disease, basal estradiol was low or below normal in 86%. Progesterone was normal in 83%, but testosterone was high in half of the cases. The response of LH to LH-RH in patients with Cushing's disease was normal in 35%, low in 35% and high in 29% of the cases. FSH response to LH-RH was normal in 23.5%, low in 23.5% and high in 53%. In patients with adrenocortical adenoma, basal of estradiol was low or below normal, but progesterone and testosterone were normal in all cases. The response of LH and FSH to LH-RH in all patients with adrenocortical adenoma was higher than normal. In three postmenopausal women, a higher response of LH and FSH to LH-RH was seen in two cases and suppressed in one case. These data suggest that the main site of suppression of the gonadal axis in patients with adrenocortical adenoma is the gonad rather than the pituitary gland or hypothalamus, though the mechanism of hypogonadism in patients with Cushing's disease is heterogeneous.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miocamycin). Part IV-2. Toxicity of metabolites of miocamycin: acute toxicity of Mb1 in rats.

Acute toxicity studies on miocamycin (MOM), non-crystalline solid, and its metabolite Mb1 were performed in mice in the previous studies. In the present studies, we evaluated acute toxicity of Mb1 in male and female rats after single oral administration at the maximum physically applicable dose of 5,000 mg/kg. Observations were continued for 1 week after treatment. It is concluded that LD0 values of Mb1 were estimated more than 5,000 mg/kg.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miocamycin). Part IV-3. Toxicity of metabolites of miocamycin: subacute toxicity of Mb1 in rats.

Miocamycin (MOM) is a derivative of midecamycin, a macrolide antibiotic and is metabolized into 4 main metabolites of Mb1, Mb2, Mb6 and Mb12. In the previous study, LD0 values of Mb1 were estimated more than 5,000 mg/kg in male and female rats as Mb1 did not exhibited any lethal toxicity even at the maximum physically applicable dose of 5,000 mg/kg. The object of this study was to examine subacute toxicity in male and female rats after repeated p.o. administration of Mb1 for 5 weeks at a daily dosage of 125, 250, 500 and 1,000 mg/kg. It is concluded that no manifest toxicity was observed in this subacute toxicity study on Mb1 in rats.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miocamycin). Part IV-5. Toxicity of metabolites of miocamycin: acute toxicity of Mb2 in rats.

Miocamycin (MOM) is a derivative of midecamycin and is metabolized into 4 main metabolites. At previous study, LD0 values of Mb1 were estimated more than 5,000 mg/kg in male and female rats. The object of this study was to evaluate acute toxicity in male and female rats after single oral administration of Mb2, a metabolite of MOM. It is concluded that LD0 values of Mb2 were estimated more than 5,000 mg/kg.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miocamycin). Part IV-6. Toxicity of metabolites of miocamycin: subacute toxicity of Mb2 in rats.

Miocamycin (MOM) is a derivative of midecamycin, a macrolide antibiotic and is metabolized into 4 main metabolites of Mb1, Mb2, Mb6 and Mb12. At previous study, the acute and subacute toxicity of Mb1 and acute toxicity of Mb2 were performed that those metabolites did not exhibit any lethal toxicity even at the maximum physically applicable dose. The object of this study was to examine subacute toxicity in male and female rats after repeated p.o. administration of Mb2 for 5 weeks at a daily dosage of 125, 250, 500 and 1,000 mg/kg. It is, therefore, concluded that Mb2 exerted no toxic effects in this subacute toxicity.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miocamycin). Part IV-8. Toxicity of metabolites of miocamycin: acute toxicity of Mb6 in rats.

Miocamycin (MOM) is a derivative of midecamycin and is metabolized into 4 main metabolites of Mb1, Mb2, Mb6 and Mb12. In the previous studies, we estimated LD50 values of Mb6 in male and female mice after single oral administration. The LD50 values were 4,150 mg/kg in male mice and 4,000 mg/kg in female mice, respectively. In the present studies, we evaluated acute toxicity and estimated LD50 values of Mb6 in male and female rats after single oral administration. Observations were continued for 1 week after treatment. It is concluded that LD0 values of Mb6 in male and female rats, were estimated more than 5,000 mg/kg as Mb6 did not exhibit any manifest acute toxicity even at the maximum physically applicable dose of 5,000 mg/kg.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miocamycin). Part IV-9. Toxicity of metabolites of miocamycin: subacute toxicity of Mb6 in rats.

Miocamycin (MOM) is a derivative of midecamycin and is metabolized into 4 main metabolites of Mb1, Mb2, Mb6 and Mb12. It is also known that LD50 values of Mb6 were 4,150 mg/kg in male mice and 4,000 mg/kg in female mice but LD0 values in male and female rats were estimated more than 5,000 mg/kg. The object of this study was to examine subacute toxicological effects in male and female rats after repeated oral administration of Mb6 for 5 weeks at a daily dosage of 125, 250, 500 and 1,000 mg/kg. It is concluded that no manifest toxicity was observed in this study with Mb6 in male and female rats after oral administration at dosage levels of 125, 250, 500 and 1,000 mg/kg for 5 weeks.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miocamycin). Part IV-11. Toxicity of metabolites of miocamycin: acute toxicity of Mb12 in rats.

Miocamycin (MOM) is a derivative of midecamycin and is metabolized into 4 main metabolites of Mb1, Mb2, Mb6 and Mb12. In the previous studies, LD50 values of Mb12 were 5,750 mg/kg in male mice and 4,950 mg/kg in female mice, respectively. The object of this study was to evaluate acute toxicity in male and female rats after single oral administration of Mb12. Observations were continued for 1 week after treatment. It is concluded that LD0 values of Mb12 were estimated more than 5,000 mg/kg.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miocamycin). Part IV-12. Toxicity of metabolites of miocamycin: subacute toxicity of Mb12 in rats.

Miocamycin (MOM) is a derivative of midecamycin and is metabolized into 4 main metabolites of Mb1, Mb2, Mb6 and Mb12. In the previous study, LD0 values of Mb12 in male and female rats were estimated more than 5,000 mg/kg. The object of this study was to evaluate subacute toxicity in male and female rats after repeated oral administration of Mb12 for 5 weeks at a daily dosage of 125, 250, 500 and 1,000 mg/kg. It is concluded that no manifest toxic effects were caused by Mb12 even at the highest dosage level of 1,000 mg/kg/day for 5 weeks to male and female rats.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miocamycin). Part VI-2. Acute toxicity in infant rats in comparison with young adult rats.

In the present acute toxicity studies on MOM, non-crystalline solid, with infant male and female rats (5-day-old) and young adult male and female rats (5-week-old), it is confirmed as follows: LD50 values were estimated more than 5,000 mg/kg in both cases of subcutaneous and oral administrations. MOM, non-crystalline solid, did not exhibit any toxic effects similarly as previously reported with infant male and female mice and young adult male and female mice. There might be no definite age difference in toxicity between young and adult rats as LD50 values were estimated more than 5,000 mg/kg in independence upon the age. There might be no definite species difference in toxicity between mice and rats.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miokamycin). Part I-2. Acute toxicity in rats.

Miokamycin (MOM) is a derivative of midecamycin, a macrolide antibiotic isolated from a culture broth of Streptomyces mycarofaciens. The object of this study was to determine the acute toxicity in male and female rats (Wistar, 5-week-old) after single i.p., s.c. and p.o. administration of MOM, non-crystalline solid, at a dose level of 5,000 mg/kg as the maximum physically applicable dose. Observations were kept for 1 week after administration. In conclusion, no animal died during an observation period for 1 week so that the LD0 values were estimated to be more than 5,000 mg/kg in all routes of administration.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miokamycin). Part II-1. Subacute toxicity in rats.

Miokamycin (MOM) is a derivative of midecamycin, a macrolide antibiotic isolated from a culture broth of Streptomyces mycarofaciens. The objective of this study was to determine the subacute toxicity in male and female rats (Wistar, SPF, 5-week-old) after repeated oral administration of MOM, non-crystalline solid, for 5 weeks at selected dosage levels of 1,000, 2,000 and 4,000 mg/kg/day. It is concluded that the maximum non-toxic dosage level of MOM, non-crystalline solid, was 1,000 mg/kg/day but without specific toxic effects with rats when it was orally administered once daily for 5 weeks.

Toxicological studies on a new macrolide antibiotic, midecamycin acetate (miokamycin). Part III-1. Chronic toxicity in rats.

Miokamycin (MOM) is a derivative of midecamycin, a macrolide antibiotic isolated from a culture broth of Streptomyces mycarofaciens. The objective of this study was to determine the chronic toxicity of MOM in male and female rats (Wistar, SPF, 5-week-old) after repeated oral administration of MOM, non-crystalline solid, for 26 weeks at daily dosages of 62.5, 125, 250, 500 and 1,000 mg/kg. The lowest dosage level of 62.5 mg/kg/day was only applied for female rats. In conclusion, the maximum non-toxic dosage level of MOM, non-crystalline solid, is presumed to be 250 mg/kg in male and female rats with p.o. administered once daily for 26 weeks.

[Toxicological studies on a new cephamycin, MT-141. I. Its acute toxicities in mice and rats].

Acute toxicities of MT-141 were studied in mice and rats to obtain the following results. LD50 value of MT-141 by i.v. administration was 6,100 mg/kg for male mice and 5,200 mg/kg for female mice. The LD50 value by i.m. administration was 8,200 mg/kg for the males and 8,600 mg/kg for the females, respectively. The mice administered with a lethal dose of MT-141 showed abnormal syndromes such as decreased spontaneous movement, decreased rate of respiration, ataxic gait, sedative state and loss of righting reflex, followed by a decrease of body weight. Gross inspection revealed no remarkable change in the organs and tissues of mice after a treatment with a lethal dose of MT-141. LD50 value of this compound was 6,600 mg/kg for male rats and 5,700 mg/kg for female rats by i.v. administration, 8,600 mg/kg for the males and 8,550 mg/kg for the females by i.p. administration, 9,600 mg/kg for the males and 9,700 mg/kg for the females by i.m. administration and more than 15,000 mg/kg for both sexes by s.c. or p.o. administration, respectively. The rats given a lethal dose of MT-141 showed abnormal syndromes such as stepping gait, face-down position, decreased rate of respiration, ataxic gait, decreased spontaneous movement and loss of righting reflex, followed by a decrease of body weight. The rats exhibited stretching behavior when given MT-141 through i.p. route and manifested vocalization when given it through s.c. and i.m. routes. The results of gross inspection and histopathological observation suggested that high doses of MT-141 induced slight renal toxicity in rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Toxicological studies on Amfenac sodium (AHR-5850) (I) Acute toxicities in mice and rats.

Acute toxicities of Amfenac sodium (AHR-5850), new nonsteroidal anti-inflammatory agent, were studied in mice and rats. Each value of LD50 by oral, sc, im, iv and ip administration with this compound was 1190, 580, 540, 550 and 790 mg/kg for male mice and 1450, 625, 610, 630 and 710 mg/kg for female mice, respectively. Rats showed higher lethality than mice. There was no significant difference of sex in the values of LD50 for mice and rats. Movement and respiration rate followed by gastrointestinal ulcer, secondary peritonitis and systemic emaciation. These results suggest that the death is caused by secondary peritonitis and systemic emaciation due to gastrointestinal ulcer.

Acute and subacute toxicity studies on collagen wound dressing (CAS) in mice and rats.

Single administration of collagen wound dressing (CAS) made from bovine derm in the form of finely ground powders was given to mice and rats via i.p., s.c. and p.o. routes and via i.v. route in the form of physiological saline extracts and it was continuously injected into mice for 28 days via s.c. route to study its acute and subacute toxicity. Examinations were made of on general conditions, body weight, food and water consumption, hematology, serum biochemistry, organ weight, and gross and microscopic findings. Results showed no marked toxicity except for local irritation which was seen only after parenteral administration. We concluded on the basis of these animal experiments that there should be no problem in regard to safety after somewhat more extensive therapeutic application of CAS as a wound dressing in clinical practice.