Cholecystokinin stimulates gonadotropin-releasing hormone release in the monkey (Macaca mulatta).

There is evidence to suggest that the arginine vasopressin release observed in association with emesis after i.v. injection of cholecystokinin (CCK) and N-methyl-D-aspartate (NMDA) is mediated by stimulation of emetic centers in the brainstem. That the GnRH-releasing action of NMDA is also sometimes accompanied by emesis led to the suggestion that stimulation of this parvocellular system by the glutamate agonist may also be mediated in part via activation of brainstem pathways. If this is the case, then other nauseogenic agents, such as CCK, should similarly elicit GnRH release. The foregoing prediction was tested in the castrated juvenile male monkey, an experimental model characterized by the absence of spontaneous GnRH release. GnRH discharges were monitored indirectly by measuring changes in circulating LH concentrations after the responsivity of the gonadotroph had been heightened by a chronic intermittent i.v. infusion of the decapeptide. An i.v. bolus of CCK at 10 and 30 micrograms/kg BW led to a distinct discharge of GnRH accompanied by emesis or other behaviors suggestive of nausea. Lower doses of CCK (1 and 3 micrograms/kg BW) failed to significantly perturb GnRH release or cause emesis, although NMDA (5 mg/kg BW; racemic form) injected i.v. 3 h after the CCK challenge led to a robust rise in GnRH. In a parallel study, three repetitive i.v. CCK injections at 2h intervals maintained intermittent GnRH release. Pretreatment with a long-acting GnRH receptor antagonist ([AcD2Nal1,4ClPhe2,DTrp3,DArg6,DAla10]GnRH -HOAc) abolished the LH response to CCK, confirming that the action of this peptide was mediated by GnRH release. Although a direct hypothalamic site of action for these agents remains the most likely possibility, since both NMDA and CCK receptors are present in the infundibular region, the present data are consistent with the notion that CCK and, by inference, NMDA may activate GnRH release in part via the stimulation of brainstem emetic centers. Plasma GH, PRL, and cortisol concentrations were also monitored during the course of some of these experiments, and the release of these hormones was observed after the administration of either the 10 or 30 micrograms/kg BW dose of CCK.

Replacement with recombinant human inhibin immediately after orchidectomy in the hypophysiotropically clamped male rhesus monkey (Macaca mulatta) maintains follicle-stimulating hormone (FSH) secretion and FSH beta messenger ribonucleic acid levels at precastration values.

This study directly tested the inhibin hypothesis by examining the ability of replacement with recombinant human (rh-) inhibin, either alone or in combination with testosterone (T), to maintain FSH secretion and FSH beta messenger RNA (mRNA) at intact levels after orchidectomy in the hypophysiotropically clamped juvenile rhesus monkey. Thirteen male monkeys (11-21 months of age) received an intermittent i.v. infusion of GnRH (0.1 microgram/min for 3 min every 3 h). After 4-6 weeks of GnRH stimulation, 10 animals were orchidectomized, and 3 monkeys were sham castrated. Hormone replacement was initiated at castration and maintained for 4 days. Three monkeys received a combination of inhibin and T replacement, 4 monkeys received replacement with inhibin alone, and 3 monkeys received T replacement alone. A continuous i.v. infusion of rh-inhibin (832 ng/h.kg) was used to replace the testicular protein, whereas SILASTIC capsules were implanted sc for T replacement. The FSH response to castration and hormone replacement was determined by measuring circulating concentrations of this gonadotropin before a GnRH pulse and for 3 h thereafter on the day before surgery and on days 2 and 4 postcastration. Circulating immunoactive inhibin was measured by a RIA that recognizes the free alpha-subunit of inhibin as well as inhibin dimers. At the end of the study, anterior pituitaries were collected for analysis of steady state levels of FSH beta, LH beta, and alpha-subunit mRNAs. Steroid replacement alone, which produced circulating T concentrations in the upper physiological range, failed to prevent the postcastration increases in circulating FSH concentrations and pituitary FSH beta mRNA levels. In contrast, when circulating immunoactive inhibin in T-replaced monkeys was maintained at precastration levels (approximately 2 ng/ml) by infusion of rh-inhibin, FSH secretion and synthesis were held at control values. When T was omitted from combined replacement, the FSH-suppressing action of the recombinant hormone was not compromised. These results demonstrate that rh-inhibin is biologically active in the monkey, and the action of inhibin to suppress FSH synthesis and secretion does not require a concomitant action of T. Moreover, because the hypophysiotropic drive to the pituitary-testicular axis was clamped, the FSH-suppressing action of rh-inhibin must be at the pituitary.

[Mass screening of prostate in Shiribeshi area in Hokkaido--incidence and clinical symptoms of benign prostatic hypertrophy and prostatic carcinoma].

A prostate mass screening was performed in 6 towns in Shiribeshi area in Hokkaido Prefecture on 384 males who were over 50 years old. The screening included digital palpation, uroflowmetry, blood sampling for tumor markers, questionnaire for voiding disturbances and transrectal sonography. Prostatic carcinoma was found in 9 (2.3%) of 384 males. An incidence of benign prostatic hypertrophy was 15.6% in 384 males. Atypical hyperplasia was found in 4 among 35 biopsied cases. Incidence of voiding disturbance such as retardation, prolongation and nocturia significantly increased with age as well as prostate size.

[A clinical study of congenital adrenal hyperplasia].

We have compared three treatments of congenital adrenal hyperplasia (CAH) for their effect on physical development. Thirteen girls and two boys with CAH due to 21-hydroxylase deficiency were treated with three different treatments, hydrocortisone (HC), dexamethasone (DXM) and cyproterone acetate (CA). The results showed that height growth was better with HC and CA than DXM, and bone excessive maturation was more suppressed with DXM and CA than HC. A dose-dependent relationship was revealed between body weight and dose of HC. Iatrogenic obesity was found in 42.9% and 38.1% of the patients treated with DXM and HC, but none of the patients treated with CA did became obese. Physical growth was better with CA treatment than HC or DXM treatment, but CA may have a suppressive effect on the pituitary-adrenal axis observed carefully, especially on prepubertal and pubertal cases.

[Effects of anti-cancer agents on cultured rat Sertoli cells].

We performed primary immature Sertoli cell culture to investigate whether or not anti-cancer agents would have a direct effect on rat Sertoli cells. Sertoli cells, isolated from testes of 18-day-old rats, were cultured in pellets with medium for 5 days. The concentration of transferrin in cultured medium were measured as the function of Sertoli cells. The anti-cancer agents cis-diamminedichloroplatinum (CDDP), adriamycin and vinblastine were selected for this study, and added to the culture medium. CDDP decreased the level of transferrin concentration in cultured medium, namely 0.5 microgram/ml of CDDP resulted in 54.9% of the transferrin concentration in the medium compared with that without any anti-cancer agents (p < 0.05), and 1.0 micrograms/ml of CDDP produced transferrin concentrations of 57.5% and 46.2% (p < 0.05), respectively. Adriamycin (0.4 microgram/ml) and vinblastine (0.5 microgram/ml) produced transferrin concentrations of 35.2% and 31.3% in cultured medium (p < 0.05), respectively. These findings revealed that anti-cancer agents have direct damaging effects on rat Sertoli cells.

[Therapeutic efficacy of testolactone (aromatase inhibitor) to oligozoospermia with high estradiol/testosterone ratio].

To our knowledge, the action of estradiol which is produced from testosterone by aromatase on human spermatogenesis has not been fully clarified. In oligozoospermia, with high values of E2/T ratio, it is considered that the role of estradiol is suppressive to spermatogenesis. In this study, alteration of spermatogenesis was evaluated when serum estradiol levels were decreased by suppression of aromatase activity. Nine male infertile patients were treated with testolactone (Teslac: 1.0 g/day, for 3 months), one of the aromatase inhibitors. Four of them had an increase in sperm count (more than 10 x 10(6)/ml relative to base line). In endocrinological findings, serum estradiol levels and E2/free T ratio were significantly decreased after treatment. Serum free testosterone levels were significantly increased in all cases, presumably from decreased sex hormone binding globulin (SHBG) levels. These findings suggested the effectiveness of the administrated aromatase inhibitor. In particular four patients whose sperm counts were improved after testolactone treatment had high values of basal serum estradiol levels and E2/free T ratio before treatment, and these values were normalized after treatment. In conclusion we suggest that an aromatase inhibitor may be effective to male infertile patients with high serum estradiol levels.

[Therapeutic evaluation of male infertility--the increase of seminal transferrin level and the improvement in sperm concentration following administration of clomiphene citrate].

This study was designed to evaluate the functional changes of Sertoli cells following the administration of clomiphene citrate for male infertility. Sperm count and seminal transferrin level were measured before and after the treatment in 22 cases of oligozoospermia (sperm count: less than 20 X 10(6)/ml) and 14 cases of subnormal sperm count group (sperm count: 20-30 X 10(6)/ml). Clomiphene citrate was administered per os for more than 3 months consecutively in a dose of 25 mg/day. Seminal transferrin concentration increased more than 1.5 times compared with pre-treatment level in 6 cases (16.7%). Among these patients, sperm count markedly increased (20 X 10(6)/ml or more than the pre-treatment level) in 3 cases (50%) and slightly increased (10 X 10(6)/ml or more than pre-treatment) in 1 case (16.7%). In 30 cases, in which seminal transferrin level did not increase, sperm count markedly increased in 6 cases (20%) and slightly increased in 6 cases (20%). Thus, sperm count was improved more frequently in the cases in which seminal transferrin level remained elevated than the cases with no elevation of serum transferrin level. Serum FSH level of patients whose seminal transferrin level remained elevated after the treatment was significantly higher than that of patients with no elevation of serum transferrin level (mean +/- SD = 32.8 +/- 18.0 mIU/ml v.s. 14.4 +/- 11.7 mIU/ml, respectively). These data suggested that the activation of Sertoli cells may contribute to the increase of sperm count following the administration of clomiphene citrate and the elevated seminal transferrin secretion may be related to the increase of serum FSH level following this treatment.

[Recent progress in surgical management for urogenital cancers--improvement of patients' QOL following surgery].

We report our experience of urinary reconstruction with ileal neobladder, and nerves-paring radical cystoprostatectomy or radical prostatectomy in an attempt to improve quality of life (QOL) in patients following surgery. While the ileal neobladder is not always indicated for patients who require cystectomy for invasive bladder carcinoma, voiding through the urethra in this urinary reconstruction contributes to improvement of post-cystectomy QOL. Daytime continence is achieved in 90% of patients. However, nighttime incontinence was found in 30%, which indicates a need for further refinement of the operative procedures or more investigations into the mechanism of continence in this condition. Recovery of erectile function following nerve-sparing radical cystoprostatectomy or prostatectomy was found in 50% of the patients who received this procedure. Subjective and objective erectile capacity before operation partly determine post-operative erectile function. Several patients whose post-operative erectile function was recovered had not had sexual intercourse after surgery. Anxiety about the disease, still inadequate penile rigidity and a non-cooperative attitude of the sexual partner were involved in the results, suggesting that more careful care and counseling before and after operation are required.

[Nocturnal luteinizing hormone secretion patterns at the onset of puberty measured using a highly sensitive immunofluorometric assay].

Nocturnal LH pulse was analyzed in 4 prepubertal, 3 early pubertal and 2 midpubertal boys by measuring serum LH concentrations. Blood samples were taken every 30 minutes while awake, and every 20 minutes after subjects had gone to sleep. Serum LH was measured by using immunofluorometric assay (IFMA) with a lower detection limit of 0.02mIU/ml. Pulsatile LH secretion during sleep was detected in all subjects examined. LH pulse amplitude had the tendency to increase with the advance of pubertal stage. After the onset of puberty, LH pulse amplitude was significantly larger than before. The LH pulse was also detected while awake in 4 (1 prepubertal, 2 early pubertal and 1 mid-pubertal) boys. LH pulse frequency was almost the same in each pubertal stage. Serum testosterone levels gradually increased with the progress of pubertal stage. These results suggest that 1) LH is secreted in a pulsatile manner in the prepubertal subjects as well as in the pubertal subjects, 2) the increase of LH pulse amplitude plays an important role for the onset of puberty, and 3) LH pulse amplitude could be a biological marker that indicates the degree of maturation of the hypothalamo-pituitary unit in prepubertal children.

Effect of kallikrein on the Sertoli cell function.

The effect of kallikrein on the Sertoli cells was investigated to determine the rat testicular androgen binding component (ABC = androgen binding protein + androgen receptor) concentration using the dextran-coated charcoal method. The results showed that the concentrations of testicular ABC were increased significantly by the administration of kallikrein to levels approaching those seen after stimulation by LH-RH or FSH. Therefore, it is suggested that kallikrein has an effect that influences the Sertoli cells directly or indirectly.

Remote and chronic access to the third cerebral ventricle of the unrestrained prepubertal rhesus monkey.

One channel of a commercially available standard-size three-channel fluid swivel was modified to permit continuous access to the brain of unrestrained prepubertal rhesus monkeys via a continuous length of small-bore Teflon tube originating from a swivel device on top of the animal's cage and terminating in the third cerebral ventricle. This system was employed to achieve continuous access to the third cerebroventricle in four monkeys for periods of up to 12 mo. The value of the system for studies of the neurochemical control of hypothalamic-releasing factor secretion was established by monitoring adenohypophysial responses to neurotransmitter receptor agonists infused into the third ventricle. Specifically, repetitive infusions of morphine (30 micrograms/infusion) elicited a robust train of prolactin discharges, and third ventricular administration of N-methyl-DL-aspartic acid (NMA; 20 micrograms) resulted in striking discharges of LH.

Magnetic resonance imaging of the male pelvic floor: the anatomical configuration and dynamic movement in healthy men.

The male pelvic organs and the rhabdosphincter were visualized using magnetic resonance imaging (MRI) in five young male volunteers (mean age, 25.6 years). The prostate was crescent-shaped in three subjects and doughnut-shaped in two subjects. The external urethral sphincter (EUS) was located anterior to and lateral to the urethra but was rare on the rectal side. The membranous urethral length measured 28-35 mm (average, 31.0 mm). The pubourethral portion of the levator ani embraced the urinary bladder, the prostate, and the membranous urethra, and the most distal portion of the levator was thickened. During anal contraction, the EUS became thinner on both coronal and sagittal images and the levator was approximated closer to the urethra. Consequently, the prostate and the bladder base were elevated and the membranous urethra was elongated by 0-12mm (average, 5.6 mm). On sagittal images, the prostate, the membranous urethra, and the rectum were pulled closer to the pubic bone by anal contraction and the retropubic area became narrower. This study clearly demonstrated that MRI was useful in examining the anatomical configuration of the male pelvic floor and its dynamic movement during anal contraction.

Role of the hypothalamic opioidergic system in the control of gonadotropin secretion in elderly men.

The role of opioidergic system in controlling gonadotropin secretion in elderly men still remains uncertain. In the present study, we attempted to examine the opioidergic inhibitory tone imposed on GnRH release mechanism in elderly men by monitoring the plasma gonadotropin response to the opiate receptor antagonist naloxone hydrochloride. Six normal young men and 10 elderly men with no endocrinological diseases volunteered for the present study. In young men, plasma LH showed a biphasic increase in response to naloxone and plasma LH level during naloxone treatment was significantly higher than the mean of prenaloxone control levels. In contrast, plasma LH in elderly men was not affected by naloxone. The ratio of the peak plasma LH response during naloxone treatment to the mean pre treatment LH level (plasma peak/basal LH ratio) declined with the advance of age (r = -0.74, P < 0.005) and also correlated significantly with plasma free testosterone level (r = 0.45, P < 0.05). These data suggest (1) that the hypothalamic opioidergic tone is reduced in elderly men and (2) that the primary testicular insufficiency with the advance of age may play a major role in the decline of the hypothalamic opioidergic tone in elderly men.

Ileal neobladder for bladder substitution after radical cystectomy.

BACKGROUND: Studer's ileal neobladder is technically simple, and favorable clinical results have been reported. However, there have been only a few follow-up studies on this type of ileal neobladder. We reviewed the clinical outcomes of patients who received Studer's ileal neobladder, with a minor modification, in our institution. METHODS: Twenty-five men underwent bladder reconstruction with Studer's ileal neobladder after radical cystectomy. The function of the ileal neobladder and voiding status were evaluated during follow-up. The follow-up period ranged from 3 to 42 months (mean, 24 months). RESULTS: The ileal neobladder achieved a large capacity at a low basal pressure, associated with a relatively low complication rate directly related to the neobladder. At 12 months after surgery, daytime and nighttime continence rates were 90% and 74%, respectively. In 5 patients who were incontinent, the maximal urethral closure pressure was statistically lower than in patients with continence. CONCLUSION: Studer's ileal neobladder is an easy operative procedure for bladder substitution, and has a relatively low complication rate directly related to the neobladder. Good compliance of the pouch and preservation of the external sphincter mechanism are the most important factors for achieving urinary continence, after construction of the ileal neobladder.

Voiding dysfunction in ileal neobladder.

PURPOSE: We attempt to understand better the voiding mechanism and determine the causes of voiding dysfunction in ileal neobladder patients. MATERIALS AND METHODS: The voiding condition in 12 bladder cancer patients who had undergone neobladder construction after radical cystectomy was examined by pressure-flow study and voiding cystourethrography. The patients were classified into 2 groups based on the maximum flow rate in a pressure-flow study: fine voiders (6), with a maximum flow rate of 15 ml. per second or more, and poor voiders (4), with a maximum flow rate of less than 15 ml. per second. RESULTS: Maximum flow rate in the fine voiders was 26.2 +/- 8.7 ml. per second (mean plus or minus standard deviation) compared with 9.4 +/- 0.9 ml. per second in the poor voiders. The fine voiders strained 1.5 +/- 0.9 times to void 100 ml. of urine, whereas the poor voiders strained 7.0 +/- 6.3 times. The bladder capacity, vesical opening pressure and vesical pressure at the maximum flow rate were similar in both groups. Pelvic floor electrical activity was amplified when the neobladder was either full or close to its capacity, and it was relaxed in coordination with abdominal straining in all patients. Voiding cystourethrography revealed that, in fine voiders, the neobladder outlet was funnel-shaped when the bladder was full and opened wider when the patient strained to void. However, in poor voiders, the neobladder outlet did not funnel even on abdominal straining. CONCLUSIONS: Neobladder patients void by abdominal straining and relaxing the pelvic floor musculature. To obtain smooth voiding in such patients, it is crucial to locate the neobladder opening at the most caudal portion and preserve neobladder outlet elasticity.

Transitional cell carcinoma of the bladder or renal pelvis in children.

Transitional cell carcinoma (TCC) of the bladder or renal pelvis is rare in children. We report 2 children with TCC of the bladder and 1 with TCC of the renal pelvis. One of the two children with bladder carcinoma experienced frequent intravesical recurrences, which is in contrast to the usual clinical course of bladder carcinoma in children. In the 3rd child, renal pelvic carcinoma was found incidentally in a renal pelvis specimen removed during pyeloplasty.

[Salivary testosterone levels in normal boys at puberty].

Salivary testosterone (Salivary-T) was measured in 133 normal boys at puberty (8-14 years old) and 21 adult males (24-36 years old) by using radioimmunoassay. Also, a simultaneous measurement was performed on salivary-T and serum total testosterone (Total-T) and free testosterone (Free-T) in order to study the correlation between saliva and serum. The results were summarized as follows: (1) Good correlations were observed in the values between Salivary-T and Total-T or Free-T. The correlation coefficient value was 0.54 between Salivary-T and Total-T, 0.84 between Salivary-T and Free-T when measured by DPC kit. Correlation coefficient value was 0.59 between Salivary-T by Wien kit and Total-T by DPC kit, also 0.77 between Salivary-T by Wien kit and Free-T by DPC kit. (2) Salivary-T, in the mean level, increased by age. The most rapid increase of Salivary-T was observed at 13 years of age. (3) Diurnal change of Salivary-T was observed at 13 years of age through adult age. Salivary-T level was highest in the morning and declined toward evening. (4) These results suggest that Salivary-T measurement was highly reliable and applicable for use in the monitoring of androgen status.

Morphological and pharmacological characterization of guinea-pig prostatic smooth muscle cells in vitro.

BACKGROUND: Prostatic smooth muscle is thought to play a major role in the pathogenesis of bladder outlet obstruction in patients with benign prostatic hypertrophy. However, the physiology of prostatic smooth muscle cells remains largely unknown, in part due to the lack of a suitable model system. We therefore sought to establish an in vitro culture of guinea pig prostatic smooth muscle cells. METHODS: Immature guinea pig prostate was treated by enzymatic digestion and the cells obtained were used to initiate the primary culture. After 3 to 4 passages, cultured smooth muscle cells were examined morphologically by immunocytochemistry and electron microscopy. The contractile properties of cultured smooth muscle cells were also examined. RESULTS: The cultured prostatic cells demonstrated hill and valley morphology, which is a hallmark of smooth muscle cells in vitro, and stained positively for desmin. In addition, electron microscopic examination of ultrastructural morphology revealed myofilaments. Confluent cultures of prostatic smooth muscle cells showed a clear, dose-dependent contractile response to phenylephrine. Furthermore, contraction of the prostatic smooth muscle cells by 10(-6) mol/L phenylephrine was completely inhibited by pretreatment with 10(-6) mol/L terazosin. CONCLUSIONS: An in vitro culture of prostatic smooth muscle cells was established. This culture is likely to provide a powerful tool for elucidating the physiology and pathophysiology of prostatic smooth muscle.

Ectopic ureterocele with a horseshoe kidney in an adult.

A rare adult case of a left ectopic ureterocele associated with a duplex horseshoe kidney is reported. To the best of our knowledge, only one pediatric case of horseshoe kidney with an ectopic ureterocele has been reported. The present case was successfully treated by ureteropyelostomy, upper ureterectomy and unroofing of the ureterocele. The patient is currently followed with excretory urograms and renograms.

Restoration of testicular transferrin, insulin-like growth factor-1 (IGF-1), and spermatogenesis by exogenously administered purified FSH and testosterone in medically hypophysectomized rats.

To investigate the relation between testicular transferrin and insulin-like growth factor-1 (IGF-1) secreted by Sertoli cells and the differentiation of germ cells in the rat, testosterone and/or purified FSH was administered to rats medically hypophysectomized by an LH-RH agonist. Spermatogenesis was quantitatively analyzed and concentrations of intratesticular transferrin and IGF-1 were measured by radioimmunoassays. The last step of spermatogenesis was quantitatively restored by combined administration of testosterone and purified FSH. Intratesticular IGF-1 concentrations were significantly increased by combined administration of testosterone and purified FSH. From these observations, it is surmised that IGF-1 may have a stimulatory effect on the last step of spermatogenesis.