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Effect of neonatal castration on liver tumor induction by N-2-fluorenylacetamide in suckling BALB/c mice.

BALB/c mice were castrated at 2 days of age and control animals were sham-operated. Untreated male and female mice were also included for comparison. One-half of the mice in each group were fed on alternate days with 1.5% N-2-fluorenylacetamide suspended in 1% gelatine by stomach tube beginning at 1 week of age for a total of 14 feedings. The experiment was terminated when the mice reached one year old. Approximately 30% of the male mice, but none of the females, developed liver tumors in groups fed carcinogen. This male predominance in the incidence of hepatocellular carcinomas was completely abolished when the animals were castrated neonatally. No lesions were observed in the liver of mice treated with gelatine suspension alone except one with neoplastic nodule. Although it has been observed that liver tumors are produce more readily in younger than in older mice, the present investigation shows that male hormonal environment during early life is more important than the age on the development of liver tumors initiated by carcinogen.

2-Acetylaminofluorene↗

Influence of testosterone, estradiol-17 beta and dihydrotestosterone on circulating LH and FSH in castrate male guinea pigs.

The influence of testosterone, dihydrotestosterone, estradiol-17 beta or combinations of these steroids on gonadotropin regulation was examined in castrate male guinea pigs. Physiological replacement therapy with testosterone prevented the postcastration rise in LH. Treatment with dihydrotestosterone and the combination of estradiol-17 beta plus testosterone was also effective. In contrast, these treatments reduced but did not prevent the postcastration rise of FSH. The negative feedback effects of these steroid treatments was not altered when 3, 6 or more than 8 weeks elapsed between castration and the initiation of steroid replacement therapy. Our data indicate that the guinea pig testosterone and dihydrotestosterone are effective regulators of LH but that other hormones may be important in the regulation of FSH.

Animals↗

Complete suppression of plasma follicle-stimulating hormone in castrated male and female rats during continuous administration of porcine follicular fluid.

The effects of continuous i.v. infusion of porcine follicular fluid (PFF), presumed to contain inhibin activity, on plasma gonadotropin levels were investigated in adult male and female Sprague-Dawley rats. One group each of male and female rats was castrated just prior to the start of treatment of 10-day duration (ACUTE). Another ACUTE group of females was treated for 30 days. A final group of female rats was ovariectomized 3 months before the start of treatment of 10-day duration (CHRONIC). When administered at a dosage of 1.0 mg/100 g body weight/24 h, charcoal-extracted PFF suppressed plasma follicle-stimulating hormone (FSH) concentrations to hypophysectomy levels within 48 h and maintained these levels for 10 days in all groups treated. In the ACUTE group of female rats treated for 30 days, FSH suppression was maintained during infusion up to 20 days. This inhibition became attenuated after 25 days of treatment and completely disappeared by 30 days. There was no discernible effect of PFF on luteinizing hormone (LH) concentrations at any time. Upon cessation of PFF treatment at 10 days in both ACUTE and CHRONIC ovariectomized rats, FSH levels rebounded to normal ovariectomy control concentrations within 2-4 days. Porcine serum (PS) was similarly extracted and administered as a control substance. It exerted no detectable effect on either FSH or LH concentrations in ACUTE or CHRONIC castrated rats. These results demonstrate that PFF alone is capable of maximally suppressing FSH secretion when administered by continuous infusion. This finding is especially significant considering that such a pronounced effect was obtained in the total absence of gonadal steroids.

Animals↗

Effectiveness of cyproterone acetate in achieving castration and preventing luteinizing hormone releasing hormone analogue induced testosterone surge in patients with prostate cancer.

PURPOSE: To our knowledge this study represents the first analysis monitoring the efficacy of cyproterone acetate (CPA) monotherapy for achieving castrate testosterone levels prior to administering a luteinizing hormone-releasing analogue (LHRHA) for treating prostate cancer in the prostate specific antigen (PSA) era. MATERIALS AND METHODS: Patients with untreated locally advanced or metastatic prostate cancer were recruited prospectively. Treatment involved a 28-day course of oral cyproterone acetate and LHRHA depot injection on day 14. Patients had serum PSA, luteinizing hormone and testosterone monitored at intervals during a 56-day period. RESULTS: A total of 15 patients with a mean age of 74 years completed the study. Near castrate serum testosterone was achieved on day 7 (mean +/- 95% CI 83.38 +/- 17.87 ng/dl). There was a significant testosterone increase after LHRHA administration on day 14 compared with the level of 160.23 +/- 36.60 ng/dl on day 16 (p <0.01). Serum luteinizing hormone mirrored testosterone, increasing from a mean of 4.93 +/- 0.61 to 15.4 +/- 6.12 nmol/l after LHRHA administration (p <0.01). Mean serum PSA demonstrated a decrease from 199.25 +/- 6.12 microg/l at day 0 to 43.77 +/- 33.08 microg/l by day 56. There was no increase in serum PSA after LHRHA administration. CONCLUSIONS: Two weeks of priming with CPA does not eliminate the surge in serum testosterone (testosterone flare) upon LHRHA administration but the testosterone increase does not exceed pretreatment levels. Furthermore, 2 weeks of CPA may not offer a benefit over 1 week in lowering serum testosterone. Finally, there is no increase in serum PSA when LHRHA is administered after priming with CPA.

Aged↗

Vas deferens and seminal vesicles epithelium after castration: a SEM study.

The effects of orchiectomy have been observed, with SEM, on the vas deferens and seminal vesicles epithelium of 20 adult albino rats. The inner surface of castrated rat vas deferens shows a reduction in height and thickness of the mucosal folds and, at the cellular apex, a remarkable decrease of the microvilli as well as the disappearance of the secretory blebs. At the level of the seminal vesicles also, castration affects both the mucosal folds, which are flattened, and the cellular apical features, with the reduction of the microvilli and the disappearance of the secretory granules. Such changes give evidence in favour of a decrease in the absorptive capacity and a suppression of the secretory activity of these epithelia, as a consequence of the fall of androgen levels.

Animals↗

Effect of castration on lectin staining in rat epididymis.

Seven rhodamine-conjugated lectins (PNA, RCA I, Con A, WGA, UEA I, SBA, DBA) were used to follow the staining pattern of the rat epididymis at different time points after castration. The affinity of the intratubular sperm mass for the lectins increased rapidly with concurrent augmentation of the staining in the principal cells but a decline of the reaction in the light cells. The light cells showed some differences in their response to castration, which was compatible with secretory/absorptive activity in caput and absorptive activity in cauda. The active phase of sperm mass destruction and epithelial involution was accompanied by local accumulation of macrophages and round cells, which also acquired an increased affinity for most of the lectins. It is concluded that the androgen-deprived epididymis is rapidly programmed for autolytic and phagocytic processes, which include the destruction of macromolecules including glycoproteins of the spermatozoa.

Animals↗

Comparison of morphine and butorphanol as pre-anaesthetic agents in combination with romifidine for field castration in ponies.

OBJECTIVE: The aim of this study was to compare two different alpha2 agonist-opioid combinations in ponies undergoing field castration. STUDY DESIGN: Prospective double-blind randomized clinical trial. ANIMAL POPULATION: Fifty-four ponies undergoing field castration. MATERIALS AND METHODS: The ponies were randomly allocated to receive one of three different pre-anaesthetic medications [intravenous (IV) romifidine 100 microg kg(-1) and butorphanol 50 micro kg(-1); romifidine 100 microg kg(-1) and morphine 0.1 mg kg(-1) IV, or romifidine 100 microg kg(-1) and saline IV] before induction of anaesthesia with ketamine 2.2 mg kg(-1) IV. Further doses of romifidine (25 microg kg(-1)) and ketamine (0.5 mg kg(-1)) were given when required to maintain anaesthesia. Quality of sedation, induction of anaesthesia, maintenance of anaesthesia, recovery, and surgical condition were assessed using a visual analogue scale scoring system and compared. The effects of the different drug combinations on heart and respiratory rate were evaluated and the recovery time was recorded. RESULTS: Anaesthesia was considered adequate for surgery in all ponies. No anaesthetic complications were observed. Quality of sedation was significantly better in the butorphanol group compared with the control group (p = 0.0428). Overall quality of anaesthesia was better in the butorphanol group compared with morphine (p = 0.0157) and control (p < 0.05) groups. Quality of induction of anaesthesia and recovery were not significantly different between groups, nor were the surgical conditions, recovery time and the number of repeated anaesthetic doses required during the procedure. Muscle twitches were observed in both the control and morphine groups. Maintenance of anaesthesia was judged to be smoother in the butorphanol group compared with the morphine and control groups (p = 0.006). Heart rate decreased significantly (p < 0.01) in all groups after administration of sedatives but did not differ significantly between groups at any time point. CONCLUSION: The combination of butorphanol and romifidine was found to provide better sedation compared with the other drug combinations. CLINICAL RELEVANCE: The combination of butorphanol and romifidine provided better sedation, but morphine was found to be a suitable alternative to butorphanol. Use of morphine and butorphanol in combination with alpha2 agonists should be further investigated to assess their analgesic effects.

Analgesics, Opioid↗

Melatonin binding sites in the Harderian gland of Syrian hamsters: sexual differences and effect of castration.

The presence of specific melatonin binding sites in the Harderian gland of Syrian hamsters was studied using [125I]melatonin. Saturation binding experiments conducted with [125I]melatonin at 37 degrees C using Harderian glands of both male and female Syrian hamsters revealed a single nanomolar-affinity site. The dissociation constants (Kd) were 6.47 and 6.94 nM for males and females, respectively. The concentration of the binding sites was 7.58 fmol/mg protein for males and 13.50 fmol/mg protein for females. Castration of male hamsters resulted in a significant increase in [125I]melatonin binding sites while chronic melatonin administration did not modify the binding properties. The results confirm the presence of melatonin binding sites in the Harderian glands of rodents. The gender-associated differences found together with the effects of castration in male hamsters suggest an androgenic control in [125I]melatonin binding sites of the Syrian hamster Harderian gland.

Animals↗

A further contribution to the question of trophic and hormonal influences on the noradrenaline content of the male reproductive tract: effect of combined androgen and estrogen treatment of prepuberally castrated rats.

58 days old, prepuberally castrated, male rats were treated with estrogen, testosterone or a combination of both hormones for 18 days. The total noradrenaline (NA)-content of the male accessory glands was increased by the hormone treatments. This increase was smallest in the estrogen treated group and largest in the group treated with both estrogen and testosterone. The NA-concentration, however, followed the reversed picture i.e. it was largest in untreated castrated rats and smallest in the rats receiving both estrogen and testosterone. Neither treatment brought the NA-amount up to the level of uncastrated controls of the same age although the weights of the secondary sex glands of rats receiving both estrogen and testosterone exceeded those of uncastrated controls. Similar, but less definite changes were observed in vas deferens and cauda epididymidis. It is concluded that a developing target area exerts some trophic influence on the adrenergic neurons innervating it. However, the trophic influence exerted by the effector organ on the nerves is not of the kind, that the innervation density and NA-concentration of the organ always are maintained at a constant level.

Animals↗

Involvement of cholinergic nicotine-like receptors as modulators of amine turnover in various types of hypothalamic dopamine and noradrenaline nerve terminal systems and of prolactin, LH, FSH and TSH secretion in the castrated male rat.

The effects of high repeated subcutaneous doses (4 X 2 mg/kg) of nicotine have been evaluated on dopamine (DA) and noradrenaline (NA) levels and turnover in the long-term castrated male rat using catecholamine (CA) fluorescence histochemistry in combination with quantitative microfluorometry. The CA turnover was evaluated by studying the decline of the CA stores following tyrosine hydroxylase inhibition using alpha-methyltyrosine methyl ester (H 44/68). In the same experiments trunk blood was collected for the determination of serum prolactin, LH, FSH and TSH levels using standard radioimmunoassay procedures. The nicotine treatment produced a significant depletion of CA stores and an increase of CA turnover in DA and NA nerve terminals of the median eminence and in peri- and paraventricular NA systems. These effects were significantly counteracted by pretreatment with mecamylamine. Nicotine significantly reduced serum prolactin and TSH levels, and after H 44/68 it also reduced LH and FSH serum levels. These actions were counteracted by mecamylamine pretreatment, except the effects on serum TSH levels after H 44/68, which were even enhanced by pretreatment with mecamylamine. Overall intraindividual correlations showed a significant correlation between reduced CA turnover in several hypothalamic areas and increased serum LH and FSH levels, increased NA turnover in the paraventricular hypothalamic nucleus and increased serum TSH levels, and reduced DA turnover in the median eminence and increased serum LH levels. It is suggested that in the castrated male rat nicotine can activate cholinergic nicotine-like receptors facilitating DA and NA turnover and release in various hypothalamic CA nerve terminal systems including those inhibiting the secretion of prolactin and LH (DA terminals in medial and lateral palisade zone, respectively) and facilitating secretion of TSH (NA terminals in the parvocellular part of the paraventricular hypothalamic nucleus).

Animals↗

Inputs to testosterone-sensitive stria terminalis neurones in the rat brain and the effects of castration.

1. The inputs to cortico-medial amygdala neurones which project directly to the area of the medial preoptic/anterior hypothalamic junction were studied electrophysiologically in urethane anaesthetized male rats. 2. In experiments with fifteen male rats it was found that none of these neurones was responsive to electrical stimulation of the ipsilateral olfactory bulb or accessory olfactory bulb or odour stimulation. 3. Experiments with four rats showed that electrical stimulation of the lateral portion of the contralateral fimbria excited 81% of these cortico-medial amygdala neurones. Their typical response to stimulation of the contralateral fimbria was a single action potential followed by an inhibitory period (20-100 ms). 4. Analysis of the polarity of evoked waves in the amygdala suggested that the fimbria input terminated in the basolateral nucleus of the amygdala and that this nucleus subsequently projected to the cortico-medial amygdala. The fimbria input was found to be contralateral in origin, crossing the mid line in the anterior fornical commissure. 5. In a further experiment 118 identified cortico-medial amygdala neurones were recorded from twelve rats (six gonadally intact and six castrated). Castration significantly decreased the percentage of these neurones responding to stimulation of the ipsilateral fimbria (20 vs. 97%) and lengthened post-excitatory inhibitory periods. 6. Results are discussed with respect to the initial finding by Kendrick & Drewett (1979) of testosterone-sensitive absolute refractory periods in cortico-medial amygdala neurones.

Afferent Pathways↗

Neonatal castration: influence on neural organization of sexual reflexes in male rats.

Most male rats castrated 4 days after birth and given exogenous testosterone in adulthood were sexually motivated but incapable of completing the mating sequence with an ejaculatory response. When tested for sexual reflexes after spinal transection, these animals displayed impairment of genital responses. Similarly treated 12-day castrates exhibited a complete mating sequence and had normal sexual reflexes. Thus neonatal testicular androgen appears to have an organizational influence at the spinal level on neural tissue mediating sexual reflexes.

Animals↗

DNA synthesis in the anterior pituitary of the male rat: effect of castration and photoperiod.

Castration increased incorporation of tritiated thymidine into total DNA in the anterior pituitary gland. Furthermore, there was a threefold increase in the percentage of labeled basophils 1 month after castration. Exposure of rats to constant light or dark also changed DNA synthesis; these changes depended on age of the animal and on exposure length. The results reflect physiologically induced mitotic activity in specific classes of pituitary cells and further suggest that neuroendocrine mechanisms may be involved in control of cell turnover in the gland.

Animals↗

Copulation in castrated male rats following combined treatment with estradiol and dihydrotestosterone.

Castrated male rats injected daily with 2 micrograms of estradiol benzoate (EB) combined with 200 micrograms of dihydrotestosterone propionate (DHTP) displayed masculine mating behavior which was indistinguishable from that of other castrates treated with 200 micrograms of testosterone propionate (TP). Significantly less copulation was seen in rats treated with either 4 micrograms of TP plus 200 micrograms of DHTP or 2 micrograms of EB. Mating in male rats may depend on the action of both estrogenic and 5alpha-dihydro metabolites of testosterone.

Animals↗

Castration of horses and donkeys with first intention healing.

A simple technique for castration of adult horses which results in first intention healing was devised. The technique involves a 'closed' castration with or without a transfixed ligature on the spermatic sac and suturing the scrotal skin with polyglycolic acid suture material. The postoperative course was characterised by mild oedema only and by rapid recovery. The results obtained during the course of removing 311 scrotal testes from horses and donkeys of all ages are described and discussed. The technique is readily applicable in the field.

Animals↗

Ontogeny of sex differences in LH and FSH levels 48 h after castration in the rat.

Serum gonadotropin levels were measured 12, 24, and 48 h after gonadectomy in male and female rats (ages, 22--60 days) to assess when during development the rate of rise of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) after castration approximates that seen in the gonadectomized adult. In females serum LH levels 48 h after ovariectomy were increased above sham levels only when the ovaries were removed prior to vaginal opening. Ovariectomy on the day of vaginal opening or at older ages resulted in no increase in LH levels by 48 h after surgery. Serum FSH levels at 24 and 48 h after ovariectomy declined with increasing age at the time of ovariectomy. In males serum LH levels at 48 h after castration increased with increasing age at the time of gonadectomy. Serum FSH levels at either 12, 24, or 48 h after orchidectomy did not change appreciably with age at the time of surgery. It is concluded that the acute pituitary secretion of gonadotropins after removal of testes in the immature male resembles that seen in the mature male early in the course of the development of sexual maturity. In contrast, the acute pituitary secretion of gonadotropins after removal of the ovaries in the immature female does not resemble that seen in the ovariectomized adult until she is mature and capable of ovulating. Thus, the observed delay in the rise of LH seen in ovariectomized adults may be a function of some aspect of the hormonal changes associated with the estrous cycle.

Age Factors↗

Effect of sex hormones on blood pressure and vascular connective tissue in castrated and noncastrated male rats.

Aortic collagen and elastin were quantitated in three groups of castrated and two groups of noncastrated male rats treated by intramuscular injection for 3 wk with oil, testosterone, or estradiol. The greatest differences were found between the castrated rats receiving testosterone and those receiving estradiol, the estradiol-treated rats having significantly lower total collagen, percent collagen, total elastin, and collagen/elastin (C/E), and higher percent elastin than those rats receiving testosterone. In noncastrated rats, administration of estradiol resulted in significantly lower total collagen, percent collagen, total elastin, and C/E. Systolic blood pressure was highest in rats receiving testerone and lowest in rats receiving estradiol. It is concluded that 1) estradiol in the presence or absence of testosterone decreases total accumulation of vascular connective tissue and alters the proportions of collagen and elastin so that the vessel is more distensible, 2) testosterone has an opposite but less marked effect than estradiol on vascular connective tissue, and 3) estradiol and testosterone alter blood pressure in opposite directions in the male rat.

Animals↗

In vitro secretion of gonadotropin-releasing hormone (GnRH) and [hydroxyproline9]GnRH from the rat hypothalamus exhibits a differential sensitivity to castration and second messengers.

The decapeptide [hydroxyproline9]GnRH (HypGnRH) has been characterized as an endogenous posttranslational product of the gonadotropin-releasing hormone (GnRH) precursor in a wide range of mammalian brains. Despite consistent biological effects, its secretion by the hypothalamus remains hypothetical. We report here in vitro secretion of HypGnRH and GnRH by the hypothalamus from intact and castrated male rats and provide evidence that they are differentially regulated. Both peptides were identified by two anti-GnRH antibodies of different specificities after separation under two high-performance liquid chromatography conditions. Calcium dependency of HypGnRH release was demonstrated under stimulation with KCl in the absence or presence of Ca2+, as well as with Bay K 8644, veratridine, methoxyverapamil, or tetrodotoxin. Activation of signaling pathways involving adenylate cyclase and protein kinases A and C (PKC) induced HypGnRH release. Expression of data as percentage of release over tissue stores revealed a two- to threefold higher release of HypGnRH than of GnRH under the different modes of stimulation used, except under PKC activation which triggered a comparable recruitment of both peptides. Castration selectively affected PKC-coupled GnRH secretion which showed a twofold lesser release than in intact rats, while the HypGnRH release was unaffected. We conclude that HypGnRH and GnRH are not secreted from the hypothalamus according to the same mechanisms.

Adenylyl Cyclases↗