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Volume 40, Issue 8
Displaying 1-6 of 6 articles from this issue
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[in Japanese]1964 Volume 40 Issue 8 Pages 1054-1064
Published: November 20, 1964
Released on J-STAGE: September 24, 2012
JOURNAL FREE ACCESSDownload PDF (1031K) -
1964 Volume 40 Issue 8 Pages 1065-1094
Published: November 20, 1964
Released on J-STAGE: September 24, 2012
JOURNAL FREE ACCESSDownload PDF (2883K) -
Hiroji OKADA, Satoru OTA, Hirohiko TAKE, Hiroshi YAMAMOTO1964 Volume 40 Issue 8 Pages 1095-1098,1047
Published: November 20, 1964
Released on J-STAGE: September 24, 2012
JOURNAL FREE ACCESSDemonstration of remarkable biological effect of 3-deoxy-steroid confused the view about the mechanism of action of steroid, while it has long been believed that the oxygen function at C-3 of steroid molecule is closely related to the mechanism of steroid action.
17α-Ethynyl-4-estrene-17β-ol (ethynyl-estrenol) is one such compound which has no oxygen function at C-3 and shows remerkable progestational activity.
In the studies reported here, the conversion of ethynyl-estrenol to 17α-ethynyl-19-nortestosterone (ethynyl-nortestosterone) have been demonstrated by the incubation with rabbit liver homogenates.View full abstractDownload PDF (559K) -
Takashi HORI1964 Volume 40 Issue 8 Pages 1099-1105,1049
Published: November 20, 1964
Released on J-STAGE: September 24, 2012
JOURNAL FREE ACCESSThe reduction of 2, 3, 5-triphenyltetrazolium chloride (TTC) in the vagina of ovariectomized mice is reported by Martin (1960) as a highly sensitive index of estrogenic activity.
This paper deals with a bioassay method for estrogen when administered subcutaneously, a modification of the method of Martin (1960) who used an intravaginal injection. Mice of 30 to 32 days of age were ovariectomized. Ten days later, they were injected subcutaneously a single dose of material. Twenty-seven hours later, 1mg of TTC was applied into the vagina, and 30 minutes later, TTC reduction was measured. Under these experimental conditions, a dose-response regression was obtained between logarithm of the dose of estrogen and vaginal TTC reduction with sufficient precision over the range of 0.0075 to 0.03, μg for estradiol-17β and 0.015 to 0.24μg for estrone. A 4-point assay of estrone against estradiol-17β gave relative potencies of 0.236 in the subcutaneous test and 0.315 in the local test. In the subcutaneous test, estradiol-17β caused nearly a ten fold increase of vaginal TTC reduction over the control level when administered at the dosage of 0.03μg. Sixteen performances of the same experiment demonstrated a good reproducibility of the response.
These results indicate that the increase of vaginal TTC reduction in spayed mice is a useful index of estrogenicity, not only for the local test but also for the subcutaenous assay.View full abstractDownload PDF (749K) -
Takashi HORI1964 Volume 40 Issue 8 Pages 1106-1116,1050
Published: November 20, 1964
Released on J-STAGE: September 24, 2012
JOURNAL FREE ACCESSVarious steroids and non-steroidal compounds were injected alone or in combination with estradiol-178 into ovariectomized mice to study their estrogenic and antiestrogenic effect, using vaginal TTC (2, 3, 5-triphenyltetrazolium chloride) reducing activity of spayed mice as the index of estrogen action.
Mice of the dds strain were used. The compounds were dissolved in sesame oil or suspended in an aqueous medium, and injected once subcutaneously in the back 10 days after castration. Vaginal TTC reducing activity was measured 27 hours after injection of the compound.
The order of activity of the compounds which caused over 5 fold increase of TTC reduction may be summarized as estradiol-17β>ethynylestradiol>diethylstilbestrol> estrone>17α-ethynyl-17β-hydroxy-5, 10-estren-3-one>estriol>testosterone. The effective dosage ranges of these compounds in the TTC reduction method closely agreed with those in the uterine weight assay. Most of the aromatic compounds and 17α-ethynyl-17β-hydroxy-5, 10-estren-3-one caused a marked increase of TTC reduction, showing a steep dose response curve, while estriol and testosterone showed a very shallow dose response line slope. The other uterotropic steroids such as 19-nortestosterone, 17α-ethynyl-19-nortestosterone, 17α-ethyl-19-nortestosterone, 19-nortestosterone phenylpropionate also caused a slight increase of vaginal TTC reduction with the dosages producing uterine weight increase, the intensity of which was much greater than the vaginal response. Progesterone, oxymetholone (2-hydroxymethylene-17α-methyl-17β-hydroxy-5α-androstan-3-one) and MER-25 (1- (p-2-diethylaminoethoxyphenyl) -1-phenyl-2-p-methoxyphenyl ethanol) also caused only a slight increase of TTC reduction with the dosage of 3 mg or more, which produced a considerable increase of uterine weight. These results indicate that the increase of vaginal TTC reducing activity closely reflects the estrogenicity of various substances.
In the anti-estrogenic activity test, the compounds were administered simultaneously in combination with 0.03, μg of estradiol-17β by a single injection at separate sites on the back. The endpoint was the percent inhibition of TTC reduction against 0.03μg of estrogen response. The order of anti-estrogenic activity of the compounds producing over 30% inhibition with statistically significance may be summarized as 17α-ethyl-19- nortestosterone>19-nortestosterone phenylpropionate>17α-ethynyl-19-nortestosterone> dexamethasone > 19-nortestosterone > estriol > progesterone testosterone > testosterone propionate. The most active was 17α-ethyl-19-nortestosterone, which caused 78% inhibition with a dosage of 300μg. In the other steroidal compounds, a dosage of 300μg or more was required to produce maximal inhibition, the percentage of which was 60% or less. In contrast to these steroidal compounds, MER-25 almost completely inhibited the estrogen action although, relatively, a large dosage was needed. These results suggest that the vaginal TTC reduction in spayed mice is a useful parameter not only for estrogen assay but also for anti-estrogen assay.
The coexistence of estrogenicity and anti-estrogenicity in most of the compounds demonstrated in the vaginal TTC reduction was discussed.View full abstractDownload PDF (1319K) -
Studies on Exophthalmos Producing SubstanceHiromasa TORIBE1964 Volume 40 Issue 8 Pages 1117-1138,1052
Published: November 20, 1964
Released on J-STAGE: September 24, 2012
JOURNAL FREE ACCESSIn order to obtain some information about endocrine exophthalmos, the author attempted to determine the exophthalmos producing activity of desiccated thyroid (thyradine), thyroxine (T4) and triiodothyronine (T3), to separate exopthalmos producing substance (EPS) from thyroid stimulating hormone (TSH) with column chromatography using pig thyrotropin preparation (pretiron) as the starting material and finally to test the effect of the thyroid hormone preparations on the exophthalmos producing response in gold fish, induced by pretiron.
The bioassay of EPS was performed by the determination of the percent increase in intercorneal distance (ICD) in gold fish after the intracloacal injection of the materials. The bioassay of TSH was performed with Bakke-Ogura's in vitro method using the suppressive effect of TSH on the release of 131I from bovine thyroid slice. Column chromatography was performed on CM-C and DEAE-C.
The following results were obtained :
1) No marked diurnal variation was observed in ICD in gold fish, and ICD was not affected by the operative procedure of cutting off the fins or intracloacal puncture.
2) As the increase in ICD after the injection of 0.9% saline was under 5%, the author defined that an increase over 5% was significant. Macroscopic exophthalmos was observed at 3 hrs. after the injection of 5 Junkmann-Schoeller Units (JSU) of pretiron, and the peak of percent increase was reached at about 5 hrs.. The exophthalmic response of gold fish was decreased in summer when water temperature showed above 30°C.
3) At 5 hrs. after the injection of pretiron a linear responsiveness of percent increase in ICD to graded doses of TSH was observed to be between 1.25 JSU and 10.0 JSU of pretiron, but on the other hand, at 10 hrs it was observed to be between 1.25 JSU and 5.0 JSU.
4) Chromatographing pretiron on CM-C using 0.01M phosphate buffer solution for the first elution, and 1.0 M NaCl for the second, the second fraction had biologically highly thyroid stimulating and exophthalmos producing activity. Changing the elute beffer gradiently from the phosphate buffer to 1.0 M NaCl, the latter half of the second fraction also showed both thyroid stimulating and exophthalmos producing activity.
5) DEAE-C, 0.005 M glycine-NaOH, 0.1 M glycine-NaOH, 0.2 M NaH2PO4 and 1.0 M NaCl were used as the elute buffer. The fractions eluted by the 1st, 2nd, 3rd and 4th buffer were named Fraction I, II, III and IV, respectively. Fraction III showed marked thyroid stimulating activity, and Fraction II, III and IV showed exophthalmos producing activity equally.
6) An exophthalmic response was observed on the 4th day following the administration of a large quantity (20 mg) of thyradine, but it was not observed following the administration of T3 or T4.
7) The pretreatment with T3 showed a marked, inhibiting effect on exophthalmos production by pretiron, compared with that of thyradine or T4.
From the results above mentioned, it is suggested that EPS is distinct from TSH and that T3 may be used as one of the therapeutic drugs for proptosis in patients with hyperthyroidism.View full abstractDownload PDF (2390K)
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