In order to evaluate a possible role of several peptides in the human urogenital tract, peptide concentrations in urogenital tissues collected from surgery were measured using specific radioimmunoassay. The specimens were extracted in boiling 0.5M acetic acid, and these extracts were utilized to measure neuropeptide concentrations, i.e., neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and peptide 7B2.
The highest concentrations of NPY were found in seminal vesicle (145±42pmol/g) and was deference (104±26pmol/g). There was no significant difference in NPY concentration between malignant and non-malignant tissues (prostate and urinary bladder). High concentrations of VIP were also observed in several urogenital tissues (seminal vesicle, was deference and urethra). VIP concentrations in prostatic cancer and carcinoma of urinary bladder seemed to be reduced, though no significant difference could be found in each corresponding tissue. Pituitary peptide 7B2 was found to be present in the human urogenital tract in relatively low concentrations. A significant difference was observed in CGRP concentration between carcinoma of urinary bladder and adjacent normal vesicular tissues (p<0.05).
These four peptide immunoreactivities were further characterized by gel permeation or high performance liquid chromatography. Each main immunoreactivity in unogenital extracts seemed to correspond to each synthetic standard or pituitary extracts (in case of 7B2). These results demonstrated that pituitary peptide 7B2 was shown to be present in the human urogenital tract and that the distribution patterns of these peptides might correlate to their pathophysiological role in the urogenital tract. Furthermore, the absence of CGRP immunoreactivity in carcinoma of urinary bladder may be useful for additional diagnostic information.

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Osteocalcin is a bone-specific protein released into the blood proportional to the rate of new bone formation. It is widely accepted that the level of serum osteocalcin is a clinical marker of bone turnover. Nephrogenic cAMP is a specific indirect parameter of the biologically active parathyroid hormone. For analysis of bone metabolism during pregnancy, we measured the concentrations of osteocalcin and nephrogenic cAMP in the maternal serum during pregnancy and in the cord serum at delivery.
Nephrogenic cAMP values (n mol/dl GF : mean±SEM) increased from the first trimester (1.5±0.21) to the term (2.11±0.11)
Osteocalcin values (ng/ml : mean ±S.D.) conversely declined from the first trimester (3.17 ±1.66) until the term (1.48±0.71) and acutely increased in the puerperium (5.91±2.58). These results might indicate that pregnancy induces a state of secondary hyperparathyroidism, but bone turnover is suppressed. In the cases of uncomplicated deliveries, the concentration of osteocalcin in the umbilical vein was significantly higher than that in the cord artery. This result suggests that a protein immunologically reactive to the osteocalcin antibody might be produced in the human placenta.

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The authors have already reported that the bone density of normal pregnant women might be kept at the same density as in normal non-pregnant women. However, it might be decreased in women with pregnancy induced hypertension (PIH) by estimating serum calcium levels, serum levels of calcium regulating hormones and calcium secretion into the urine. In order to demonstrate this theory, the degree of bone density in the second metacarpal bone of normal or PIH pregnant women was measured by X-ray using microdensitory method (MD method). In MD method, six indices, such as MCI, d, GS_min, GS_max, ∑GS/D and densitometric pattern, are calculated by computer analysis of the X-ray of the bilateral hands. By the evaluation of the degree of bone atrophy, scores such as 0 better than the regression line of healthy women, which were prepared according to each age, 1 until 1δ to the aggravation, 2 until 2δ, and 3 more than 2δ were totalized and evaluated as normal, initial stage of bone atrophy, 1st degree of bone atrophy, 2nd degree of bone atrophy and 3rd degree of bone atrophy for 0-3 scores, 4-9 scores, 10-12 scores and 13-18 scores, respectively (δ = 1 S.D.).
The metacarpal index (MCI) of normal pregnant women in 3rd trimester was more than the mean in all cases, while cases more than 2δ of the mean were noted in 29.4% of mild PIH and 11.8% of severe PIH, and a decreasing tendency of width of bone cortex was considered in PIH women. On the other hand, width of bone marrow (d) increased significantly in mild and severe PIH women. In the index for the density of only bone cortex area (GS_max) in PIH women, cases less than mean -1δ were noted in 29.3% of mild types and in 11.8% of severe types respectively, and a high incidence was noted even though it was insignificant compared with 7.4% of normal pregnant women. In the index of the densities of bone cortex and bone marrow (GSmin), cases less than mean -1δ were noted more frequently in PIH women than normal pregnant women, but in the index of bone density per unit length (∑GS/D) no differences were noted between PIH and normal pregnant women. On the other hand, in densitometric pattern, cases of more than B, the border line of healthy women (A) and early stage of osteoporosis (C), were noted in 7.7% of normal pregnant women, but they were 3 5.3% in mild PIH and 47.1% in severe PIH, and a higher incidence of osteoporosis was noted in PIH women than in normal pregnant women. Moreover, a case of C, which was not recognized in normal pregnant women, was observed in PIH.
With regard to the 6 indices mentioned previously, the total score of normal pregnant women was 0-3 in all cases, and not one case judged as osteoporosis was observed. On the other hand, cases with osteoporosis in the early - 2nd stages were noted in 41.2% of mild and severe PIH pregnant women, and the incidence of the cases was significantly high in PIH.
In conclusion, the proposal of the authors in a previous report that bone density in PIH women might be decreased according to the kinetics of serum Ca concentrations and Ca regulating hormones was actually confirmed by the estimation of the bone density in X-rays.

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The peptide, 7B2, originally isolated from pituitary, has been shown to be present in endocrine tumors of high concentrations in pancreatic islet tumors. Plasma from most of these patients showed very high 7B2 immunoreactivity (IR-7B2) though there is a lack of knowledge concerning physiological and pathological changes in plasma IR-7B2 levels in other conditions. To assess whether or not there is any alteration in circulating IR-7B2 levels due to age, sex or any specific condition, plasma levels of IR-7B2 were measured in the fasting state in 106 healthy subjects aged 3 months to 91 years, 101 diabetics, 28 patients with hyperthyroidism. 7 patients with primary hypothyroidism, 13 patients with liver cirrhosis, 43 patients with chronic renal failure, 35 patients with cerebral vascular accident, and 26 pregnant subjects. Twenty-four cord bloods were also included. The responses of circulating IR-7B2 to oral glucose, intravenous arginine infusion, volus thyrotropin (TRH) or volus luteinizing hormone-releasing hormone (LH-RH) injection were also evaluated. Particularly high IR-7B2 levels were found to exist in cord blood. Postnatally the concentrations decreased gradually with age to adult values (15.6±2.9pmol/liter (mean±SE) in 20's -60's), though plasma IR-7B2 levels again increased significantly in over 70's (37.1±3.2pmol/liter; P<0.01). There was no significant difference in plasma 7B2 levels in either sex. Among the pathological conditions studied, significantly high IR-7B2 levels were observed in patients with chronic renal failure (175.1±35.9pmol/liter). Some of the pregnant patients in their third trimester also showed high plasma IR-7B2 levels. A small but significant rise in plasma IR-7B2 was observed after a glucose load in control subjects and diabetics. Intravenous LH-RH exerted a rise in plasma 7B2 concentrations though arginine and TRH showed no significant effect on plasma IR-7B2 concentrations. Compared with the plasma concentrations, ten to fifty-fold high levels of IR-7B2 were observed in cerebrospinal fluid (CSF) from patients with cerebrovascular accidents or multiple sclerosis. These results suggested that the kidney plays a major role in 7B2 degradation and that LH-RH simulates IR-7B2 release from the pituitary gland. Whether reduced clearance or increased production was responsible for the IR-7B2 elevation in subjects under 10 years or over 70 years requires investigation. Furthermore, high levels of IR-7B2 in CSF might indicate its specific role for the central nervous system.

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We administered two cholecystokinin antagonists to dogs intravenously (i.v.) and into the third cerebral ventricle (i.t.v.). Proglumide (3-300mg/kg/hr i.v. or 0.1-10mg/dog i.t.v.) reversed the satiety previously shown by mongrel dogs after i.t.v. CCK-8. A new glutaramic derivative, CR1409, blocked this satiety even more strongly when administered by either route.
Proglumide increased proglumide levels in ventricular fluid, indicating its ability to cross the blood-brain barrier. However, i.t.v. proglumide did not appear in the blood during the observation period. These results suggest that systemic proglumide and CR1409 act as antagonists of the central CCK receptor concerning satiety in dogs; intravenously administered proglumide was found to cross the blood-brain barrier and partially but significantly reverse the satiety caused by CCK-8.

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The purposes of the present study are to demonstrate the presence of atrial natriuretic peptide (ANP) in canine cerebrospinal fluid (CSF) and to determine its origin as either the brain or atrium. Fifty-seven mongrel canines weighing from 7.5 to 23.0kg (male : 28, female : 29) were anesthetized with sodium pentobarbital (30mg/mg, iv) and were ventilated with a Harvard respirator. 16 canines (11.5 to 16.0kg) were used to examine the effect of endogenously increased plasma ANP level on the ANP concentration of the CSF in acute heart failure induced by experimental aortic regurgitation. Subsequently to examine the effect of exogenously increased plasma ANP level on the ANP concentration of the CSF, physiological and pharmacological doses of synthetic human alpha-ANP were continuously infused into the right ventricle (25ng/kg/min. and 250ng/kg/min., respectively) for 32 min. in 15 canines (8.0 to 23.0kg), only physiological dose (25ng/kg/min.) was infused for 180 min. in 8 canines (12.5 to 23.0kg). The concentrations of ANP in canine CSF and plasma were measured by our highly sensitive and specific radioimmunoassay (RIA). The molecular forms in the plasma, CSF and the atrium and hypothalamus tissues were determined by gel permeation chromatography (GPC).
The ANP concentration in CSF was 2.8±1.2pg/ml (mean±SD), lower than that in the plasma which was 51.5±19.9pg/ml, and no correlation was found between them (r= 0.16, p=ns). Plasma ANP concentrations increased from 46.5±13.0pg/ml to 94.6±27.7pg/ ml according to a rise of the left atrial pressure by experimental aortic regurgitation. However, no significant change was noted from 3.7±0.7pg/ml to 3.8±1.0pg/ml in CSF ANP concentrations during the aortic regurgitation. The ANP concentration in the CSF did not change significantly while the plasma ANP concentration greatly increased following each intravenous infusion of the synthetic alpha-ANP. Only a single peak corresponding to a low molecular weight form of ANP in the position of authentic alpha-ANP in the canine CSF was observed by GPC, while there were peaks for both low and high molecular forms of ANP in the canine plasma. Furthermore, both low and high molecular weight peaks were observed for the right atrium and hypothalamus tissue extracts by GPC, and those tissues of the right atrium and hypothalamus contained ANP concentrations of 1.97ng/mg wet tissue and 2.6pg/ml wet tissue, respectively.
These results indicate the presence of ANP in canine CSF and that it does not come from blood that has seeped across the blood-CSF barriers but may originate in the brain.

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Specific antiserum for 17-hydroxyprogesterone (17-OH-P) was prepared by immunizing 7α- (2-carboxyethylthio) -17-OH-P conjugated bovine serum albumin (BSA) in rabbits. Using this antiserum, 17-OH-P enzyme immunoassay for dried blood spots on filter paper was established. As a label, alkaline phosphatase was coupled covalently with 7a-carboxymethylthio-17-OH-P by carbodiimide method. B/F separation was carried out by the addition of anti-rabbit IgG goat antiserum. All specimens used were punched out with a paper puncher of 3mm diameter. The assay sensitivity was 2pg/tube, which was estimated by two standard deviation at zero concentrations of the calibration curve. Cross reactivities of this antibody were as follows : 11-deoxycortisol (8.21%), 17-0H-pregnenolone (3.33%), progesterone (1.67%), 11-deoxycorticosterone (0.31%), cortisol (0.16%), pregnenolone-3-sulfate Na salt (0.03%), dehydroepiandrosterone (DHEA) (<0.03%), 16a-OH-DHEA (<0.03%), DHEA-3-glucuronide (<0.03%), DHEA-3-sulfate Na salt (<0.03%), pregnenolone (<0.02%). Intra-and inter-assay coefficient of variations were 4 14% and 9-18%, respectively.
In normal babies, 17-OH-P concentrations measured directly (without sample extraction) were below 23pg/disk (n=204). The histogram of 17-OH-P level in normal babies obtained by the direct method was distributed lower than that obtained by the enzyme immunoassay system (range : 4-79pg/disk, n=268) which used antibody raised against 17-OH-P-3-O-carboxymethyloxime conjugated BSA (Enzaplate, Sapporo Diagnostic Laboratory). Correlation of 17-OH-P concentrations in dried blood spots of normal babies obtained by direct method (x) and ether extracted method (y) was y = 0.67x - 0.11 (n=45). The values of 17-OH-P concentrations by the direct method were about 1.5 times higher than those obtained by the ether extracted method. However, this difference was smaller than that reported by other investigators (-10 times higher) who used antibody raised against 17-OH-P antigen with bridge site at 3 or 4 position. In immature babies, 17-OH-P concentrations on dried blood spots obtained by the direct method ranged from 6 to 189 pg/disk (n=18). Although these values were higher than the 17-OH-P values in normal babies, the values were distributed in the same range as those in normal babies determined directly by the other investigators who used antibody raised against 17-OH-P antigen with bridge site at 3 or 4 position. These results suggest that the present enzyme immunoassay using anti 7a- (2-carboxyethylthio) -17-OH-P antiserum is more specific than those so far reported. Meanwhile, in the present study, large differences in 17-OH-P values between direct method and extracted method (n=18, range : 1-29pg/disk) were observed in the dried blood spots of immature babies (max. 19 times). This may possibly be due to cross reactant substances in dried blood spots in immature babies.

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In vivo effects of prostaglandin D₂ (PGD₂) on rat testicular testosterone levels were studied. Testosterone levels were determined by HPLC after 6 hours of PGD₂ and of hCG. The testosterone levels were decreased by dose of 500μg/rat intraperitoneal injection. Testis testosterone levels were subsequently decreased by intratesticular injection with PGD₂ (1.0, ug) into the right testis. The left testis received vehicle alone for control. The increase in testosterone levels induced by hCG (50,100 and 500 unit/rat s.c.) were also inhibited by intraperitoneal injection of 500μg of PGD₂. On the other hand, intratesticular injection of hCG (0.01, 0.1 and 1.0 unit/testis) caused an increase in testosterone levels according to dose of hCG. The increase in testosterone levels by hCG was inhibited by simultaneous in-jection of PGD₂ (1.0μg/testis). These results suggest that PGD₂ may play an inhibitory action on LH action in androgen synthesis in the rat testis.

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We reported three cases of Graves' disease which showed unusual histological findings featuring solid follicles, multinucleated giant cells and diffuse infiltration of histiocytes as well as lymphocytes in the whole section of the resected thyroid. Characteristics of these three cases were as follows :
(1) Clinically, longer duration of the disease and exophthalmos were their prevailing findings.
(2) On laboratory data, antimicrosomal antibody showed extremely high titers with 100,000 to 400,000.
(3) Their operative findings were different from ordinary Graves' goiters in that colors of the goiter were yellow-red or gray-red, surface was rough and coarse, consistency was firm, and adhesions with the adjacent connective tissue were noted.
(4) Postoperative clinical outcome was quite similar to that of ordinary Graves' patients.
From these findings, these three cases were considered to be different from either so-called Hashitoxicosis or silent thyroiditis or Graves' disease with granulomatous foci, and it was suggested that these three cases might be a subgroup of Graves' disease or another hyperthyroidism than ordinary Graves' disease. Further accumulation and analysis of such cases will be necessary in order to answer this question.

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In this study, we attempt to clarify the role of sodium-potassium adenosine 5'- triphosphastase (N⁺-K⁺ ATPase) on the regulation of systemic blood pressure and renal sodium metabolism. Male Wistar rats (n=20) were divided into four groups : 1) controls (n= 5), 2) sodium loading rats (NaCl group, n=5), 3) deoxycorticosterone (DOCA group, n=5) injected rats with intact kidneys (DOCA group, n=5), 4) DOCA and ouabain-injected rats with intact kidneys (ouabain groups) and raised for six weeks. The changes in urine volume and urinary sodium were determined, and urinary excretions of norepinephrine (NE), epinephrine (E) and dopamine were measured in week six. Then blood samples were obtained from the inferior vena cava, and plasma cyclic AMP (cAMP) and cyclic GMP (cGMP) concentration were measured. At the time of sacrifice, systolic blood pressure of the ouabain group (157.3±24.8mmHg) was significantly higher than the control group (124.5±6.9 mmHg) (p<0.05), but no significant differences were observed between the control group, NaCl group (123.7±7.0 mmHg), and DOCA group (128.1±4.5 mmHg). Urine volume and sodium excretion in the NaCl, DOCA and ouabain groups all increased, but the natriuretic response in the NaCl group was smaller than in either the DOCA and ouabain groups. Urinary NE in the NaCl, DOCA and ouabain groups was 1396±1025 ng/day, 640±351 ng/ day, 607±177ng/day, respectively, and NE excretions in these groups were higher than in the control group (138±104ng/day). Urinary E in the NaCl group was higher than in the control group, but there were no significant differences among the control, DOCA and ouabain groups. Urinary dopamine in the NaCl (5723±2028ng/day), DOCA (7661±5992 ng/day) and ouabain (4077±1984 ng/day) groups was higher than in the control group (2081±483ng/day). Plasma cAMP in the DOCA and ouabain groups was slightly higher than in the control group, while no significant differences in plasma cAMP between the control and NaCl groups were observed. Plasma cGMP in the DOCA group was slightly higher than in either of the other three groups. These results suggest that Na⁺-K⁺ ATPase may play an important role in the maintenance of blood pressure through sodium efflux and that ouabain, Na⁺-K⁺ ATPase inhibitor, may change the renal sodium excretion and vascular responsiveness to endogeneous pressor substances, leading to higher blood pressure after the administration of sodium and mineralocorticoid.

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