The rapid calcium infusin test (Goldsmith, R.S. et al. 1962) was evaluated in 3 normal subjects, 6 patients with parathyroid disorders, 12 patients with urolithiasis and 12 patients with various other diseases. Twenty-five milliliter of 2% calcium chloride solution was infused intravenously over a ten-minute period starting at 9 a.m. and the response of urinary phosphate excretion to calcium infusion was investigated for 3 hours.
All normal subjects, 10 patients with urolithiasis and 8 patients with various other diseases responded to calcium infusion, but a patient with hyperthyroidism and 2 patients with urolithiasis showed delayed responses to calcium infusion. The phosphaturic rhythm was unaffected by calcium infusion in a patient with renal tubular acidosis, a patient with Fanconi's syndrome, 3 hypoparathyroid patients and a patient with multiple myeloma who had severe impairment of renal function. The abnormal spontaneous phosphaturic rhythm, i.e. a progressive fall in phosphorus excretion, was observed in a patient with acromegaly and a patient with hypercalcemia of unknown etiology under corticosteroid therapy. A hyperparathyroid patient associated with adult Fanconi's syndrome showed a decreasing Up/Ucr ratio following calcium infusion, though his spontaneous pattern was not ascertained.
The fall in urinary phosphorus excretion below the control level within 3 hours after calcium infusion may be considered as a normal response to the rapid calcium infusion test, when calcium chloride is infused instead of calcium glucoheptonate.

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Wistar系ラットを妊娠させその妊娠後期にtestosterone propionate (TP) を連続 (妊娠第15～21日;1日量0.05, 0.1, 0.5, 5.Omg/rat) 又は1回 (妊娠第15, 18, 21日;5, 10mg/rat) 皮下投与し, 出生仔性機能分化に対する影響を調べた.得られた結果を概括すると,
(1).TP連続投与群妊娠ラットの分娩は5mg投与の1例を除き正常であつたが, 1回投与群ラットでは妊娠第18日, 第15日と妊娠中期に近く投与されるほど分娩遅延が著しかつた.
(2).胚仔再吸収はTP連続投与の場合は投与量の増加につれ, 又1回投与の場合は妊娠中期に近く投与されるほど著明であつた.
(3).TP連続投与群では投与量の増加につれ, 又1回投与群では妊娠第18日投与により雌新生仔肛門-外性器間距離 (AGD) の伸長即ち男性化 (masculinization) が認められた.一方雄新生仔AGDは対照と著差を示さなかつた。このAGDに出現する変化は仔体重の変動と特に一定の関係がなく, 胚仔発育と独立した変化である.
(4).TPl日0.5mg以上連続投与並びに妊娠第18日1回投与により出現した雌ラットAGDの伸長は生育につれ対照との差がより著明となり何ら正常への回復傾向を示さなかつた.さらに出生時AGDの伸長がなかつた妊娠第21日1回投与群雌ラットも生育につれ男性化を示した.一方雄ラットAGDは生育につれ短縮の傾向を示したが投与量, 投与時期との間に一定の関係を認め得なかつた.
(5).腔開口を指標とする雌ラット性成熟, penis形態変化を指標とする雄ラット性成熟ともにTP投与群ラットは対照に比し遅延を示した.又TP1日5mg連続投与群並びに妊娠第18日1回投与群雌ラットは全く腔開口を示さず, TPO.5mg連続投与群, 妊娠第21日1回投与群雌ラットの外性器部位は形態的に異常を示した.
以上の諸結果は胎生期に投与したandrogenが末梢的に外性器部位に作用し雌胚仔男性化を生ずる以外に, 性成熟遅延現象に明らかなように中枢的に作用し性機能発現に影響している可能性を示唆するものである.

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Wistar系ラットを妊娠させその妊娠後期にtestosterone propionate (TP) を連続 (妊娠第15～21日;1日量0.05, 0.1, 0.5, 5.0mg/rat) 又は1回 (妊娠第15, 18, 21日;5, 10mg/rat) 皮下投与し, 出生仔性機能分化に及ぼす影響を性周期, 交尾能, 解剖所見などより調べた.得られた結果を概括すると, (1), TP1日0.05mg, 0.1mg連続投与群ラット及び妊娠第15日1回投与群ラットは対照と同様正常な性周期を示した.これに対しTP1日0.5mg連続投与i群並びに妊娠第21日1回投与群ラットは膣垢に角化細胞が連続して出現するいわゆる連続発情 (persistent estrus) を示すのが特徴的であつた.しかもこの連続発情はprogesterone投与によつても中断されなかつた. (2), 正常性周期を示した雌ラットの交尾能は対照と何ら差がないが, 連続発情が認められた雌ラットは殆んど交尾しなかつた.雄ラットの交尾能も対照に比し低下を示し, 特にTP連続投与群で著明であつた.雌ラットに於ける交尾能の低下は外性器部位の形態異常が大きな原因と考えられるが交尾例もあることより中枢性変化による原因を除去し得ない. (3), 生後130日目の雌ラット解剖結果ではTP投与群ラット下垂体重量が対照に比し減少を示したが卵巣, 子宮重量は対照と特に差がなかつた.一方生後160日目の雄ラット解剖結果では下垂体, 睾丸重量が対照に比し減少を示したが, 前立腺, 貯精のうなどの重量は投与形式との問に一定の関係がなく減少傾向を示すのみであつた. (4), 卵巣の組織所見は極めて特徴的であつた.即ちTP投与群でも正常性周期を示すラット卵巣は対照と同様種々発育段階の濾胞及び黄体によりしめられていた.これに対し膣垢に角化細胞が連続して出現した連続発情ラット並びに全く膣開口を示さなかつたラットでも, 黄体が無く大濾胞のみより成る卵巣及び濾胞と黄体の両者が存在する卵巣の2種が認められた. (5), 雌ラット下垂体中gonadotropin含量を調べたところ, TP投与群ラットのFSH活性は対照に比し増加の傾向を示したが著差はなく投与形式との関係も明確でなかつた.一方LH活性はTP連続投与群ラットでは投与量の増加につれ減少を示したが1回投与群ラットではこのような減少は認められなかつた.
以上の諸結果より妊娠ラットに投与したandrogenの影響は胎盤を介して胚仔に及び, 一方では中枢的に作用し胚仔視床下部の性機能分化に影響を及ぼしgonadotropinの周期的産生, 放出を支配している視床下部の機能発現を抑制し, 卵巣では黄体欠如, 濾胞の異常発育を生じ, 濾胞より分泌されるestrogenの作用により腔垢の連続角化を生ずるが, 又他方では末梢的に作用し外性器部位の男性化を生ずると同時に膣に直接作用し腔壁の細胞分化に不可逆的変化を生じその結果腔垢の連続角化を生ずるという2つの作用機序を有することが明らかである.

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Measurement of luteinizing hormone (LH) in the urine has much significance in the diagnosis of menstrual disorders. Urinary LH assay has been previously carried out by troublesome, time-consuming bioassay. Recently, immunological method has become available for that purpose, since Wide and Gemzell (1961) reported LH-immunoassay using HCG antigen-antibody system which was based upon the cross-reaction between HCG and LH. It is the purpose of this report to demonstrate the cross-reaction between HCG and LH, to establish the immunological assay method of urinary LH and to investigate the relationship between immunological and biological activites of LH.
Anti-HCG sera were obtained from rabbits received 2 injections each of 1,000 IU of crude preparation of HCG (Primogonyl, HCG-P), freshly emulsified with complete Freund's adjuvant at two weeks interval. The injections were made in the footpads and i.d. After 3 weeks of rest, a series of booster injections with 1,000 IU of HCG-P without adjuvant were given i.p. nine times at every other day. Rabbits were bled 7 days after the last injection and the sera were kept. The antibody titers were in the range from 6,400 to 51,200 by tanned hemagglutination. Antiserum which had the highest antibody titer was used in this study.
This anti-HCG serum contained several antibodies which reacted not only with HCG but also with normal human serum (NHS) and child urine protein (CUP) by immunoelectrophoresis. In order to remove antibodies against proteins other than HCG, 1 ml of anti-HCG serum was absorbed with 30mg of lyophilized NHS and 20mg of CUP, thus the absorbed anti-HCG serum showed one precipitin band only to HCG. This absorbed serum also showed one precipitin band to human menopausal gonadotropin (Pergonal, HMG-P) which contained LH and FSH by agar gel diffusion and immunoelectrophoresis, and this band completely coalosced to the band wnich appeared against HCG.
This absorbed anti-HCG serum was examined for neutralization of LH and FSH. An assay method based on the rate of increase in the weight of the ventral prostates of hypophysectomized male rats (Greep) was used for LH assay and Steelman-Pohley's augmentation method was used for FSH assay. When 2 IU or 4 IU of LH and 2 IU or 4 IU of FSH were incubated with 0.002 ml of absorbed anti-HCG serum at 37°C for 60 min., biological activity of LH was completely neutralized but the FSH activity was not. From the above immunological and biological results, it could be concluded that LH had a similar antigen to HCG and LH was completely neutralized with anti-HCG serum.
The amount of LH in the urine is so small for immunoassay that concentration of urine using pervaporation, carbowax 6,000 and alcohol precipitation to 1 : 20 is necessary. The recovery of LH was almost complete. The components of medium to dissolve the standard HCG influenced the sensitivity of tanned hemagglutination inhibition reaction. The concentration from 1 to 1 : 20 of the child urine as the medium of HCG, showed 4 fold as sensitive as phosphate buffered saline, thus 1 : 2 concentrated child urine was used for the medium to dissolve the standard HCG or to dilute the test materials.
Ten women from 23 to 34 years old who had the normal menstrual cycle, were examined every day for BBT, and LH value in the first morning urine by immunoassay. Seven women who showed clearly biphasic BBT gave the peak of LH level, in the range from 80 IU to 320 IU HCG equivalent/L, from 15 to 13 days before next menstruation. These peaks coincided with the rise of BBT.
Three women among them were examined LH level in the urine by using immunological and biological methods simultaneously. The urine for each 48 hours during the cycle war collected, and COml of that was used for immunoassay and the rest was used for bioassay. The kaolin adsorption method by Bradbury-Matsushima was used to extract gonadotropin from the urine.

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