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Effect of undernutrition on pulsatile luteinizing hormone (LH) secretion in castrate and intact male rats using an ultrasensitive immunofluorometric LH assay.

The recent development of an ultrasensitive immunofluorometric rat LH assay makes possible evaluation of pulsatile LH secretion in intact male rats under physiological conditions of minimal volume blood sampling without requiring orchidectomy. Specifically, we applied this assay to determine the effect of macronutrient restriction on pulsatile LH secretion in the presence or absence of testes. In testes-intact rats, halving of food intake for 7 days while maintaining micronutrient supply caused a reduction of mean, maximal, and basal LH levels and LH pulse amplitude (all P < 0.05) compared with those in ad libitum fed controls. The loss of body weight was positively correlated with decreases in mean LH level, pulse amplitude, and area under the curve (all P < 0.009). In contrast, the same food restriction in castrated rats caused an increase in pulse length and area under the curve and a decrease in pulse frequency, but did not change mean, maximal, and basal LH levels or LH pulse amplitude compared to castrated ad libitum fed controls (all P < 0.02). The observed positive correlations between body weight and the LH secretion parameters in intact rats were absent or reversed in castrated rats. This study demonstrates qualitatively different effects of macronutrient restriction on pulsatile LH secretion in castrated and intact rats, indicating that it is not necessary valid to extrapolate consequences of undernutrition on LH secretion from castrate to intact male rats. We conclude that undernutrition-induced inhibition of LH secretion involves both an indirect suppression of LH secretion via amplification of endogenous testicular negative feedback as well as more direct suppression of GnRH release.

Animal Nutritional Physiological Phenomena↗

Effects of sex steroids on secretory granule formation in gonadotropes of castrated male rats with respect to granin expression.

Pituitary gonadotropes show sex-related differences in their ultrastructure. Typical gonadotropes of male rats exhibit both large granules, which contain chromogranin A (CgA), and small granules, which contain secretogranin II (SgII). In contrast, typical female rat gonadotropes show only a very few large granules among the numerous small granules. To clarify the nature of the biogenesis of these secretory granules and the effects of sex steroids, the ultrastructural and immunocytochemical changes in gonadotropes were examined in castrated male rats supplied with a testosterone or estradiol implant. In castrated rats, pituitary expression and plasma levels of LH increased drastically, but the pituitary content of CgA decreased. The majority of gonadotropes then showed features of "castration cells" containing many small secretory granules. A testosterone implant to castrated rats remarkably suppressed the expression and circulating levels of LH and increased the CgA content in the pituitary to near-normal levels. In this situation, immunocytochemical studies demonstrated that gonadotropes again exhibited large and small secretory granules with the respective localization of CgA and SgII. On the contrary, in castrated rats supplied with an estradiol implant, the expression and content of CgA in the pituitary were remarkably suppressed, and large secretory granules disappeared from gonadotropes. These results suggest that the expression of CgA in gonadotropes is regulated differently by male and female sex steroids. These different effects of androgen and estrogen on the expression level of CgA are closely associated with the sex-related differences in the ultrastructure of secretory granules within gonadotropes.

Animals↗

Castration-induced apoptotic cell death in the Brown Norway rat prostate decreases as a function of age.

Growth and differentiation of the prostate gland depends upon androgens, yet overgrowth of the human prostate occurs later in life when serum levels of testosterone are declining. We have reported a similar phenomenon in the Brown Norway rat, but the age-dependent overgrowth of the prostate is confined to the dorsal and lateral lobes and, hence, is lobe specific. Because tissue growth depends upon the balance between proliferation and death of cells, the present study was designed to investigate whether cell death differed in the various prostatic lobes of Brown Norway rats as a function of age. Apoptosis of cells in the ventral, dorsal, lateral, and anterior lobes of the prostate was examined in young (4-month-old) and old (24-month-old) Brown Norway rats after castration. Whereas castration caused tissue weights of all four prostatic lobes to decrease over the course of 10 days, this occurred more rapidly and to a greater magnitude in the ventral than in the dorsal, lateral, and anterior lobes. Tissue DNA content, a measure of cell number, decreased only in the ventral lobe after castration. DNA fragmentation, indicative of apoptotic cell death, was detected by in situ labeling using the terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling method and as intranucleosomal cleavage of genomic DNA analyzed by agarose gel electrophoresis. Both methods demonstrated the correlation between loss of DNA content and apoptotic cell death in the ventral lobe, whereas only the highly sensitive terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling (TUNEL) method revealed relatively few dying cells in the dorsal, lateral, and anterior lobes after castration. Moreover, when examined as a function of age, less cell death occurred in all four lobes of old rats compared with young rats. In both young and old rat prostates, cell death was observed in epithelial and stromal cells within the ventral lobe where apoptotic cells were detected throughout the branched ductal network and were not restricted to a particular region. Taken together, these studies demonstrate the marked differences in cell death and survival between the different rat prostatic lobes in response to castration and further suggest that the androgen-sensitive apoptotic response is age dependent. Hence, the lower rates of cell death observed for the dorsal and lateral lobes, accompanied by the further decline that occurs with increasing age, are important components of the age-dependent and lobe-specific overgrowth observed for these lobes. Moreover, the age-dependent decline in apoptotic cell death observed in the prostates of old rats suggests that prostatic cells develop androgen independence as a function of age, and survival of these cells does not require androgen.

Aging↗

Effects of sex steroids on the positive estrogen feedback mechanism in intact women and castrate men.

We studied the effects of prolonged testosterone treatment on ovulatory function and positive estrogen feedback in women and of prolonged estrogen priming on gonadotropin feedback in castrate men. An estrogen provocation test was carried out in 4 groups of transsexual subjects: 12 female transsexuals in their early follicular phase (days 3-5; group 1A), 8 females who had been treated with Depo-testosterone (T) for 3-6 months (group 1B), 11 men who had been castrated 3 months previously (group 2A), and 4 male castrates treated with oral estrogen for 3 months starting 3 months after castration (group 2B). The estrogen provocation test consisted of 3 GnRH tests (100 micrograms) carried out immediately before (0 h) and 44 and 92 h after an im injection of estradiol valerate (10 mg). Responses to the estrogen provocation test in women with normal menstrual cycles (group 1A) were typically female. After initial suppression at 44 h, a LH surge (positive feedback) occurred at 92 h. Pituitary responsiveness, however, was amplified both at 44 and 92 h. Prolonged T priming of women in group 1B did not inhibit the estrogen-induced LH surge, nor was the amplitude of the surge blunted. Removal of androgens and other testicular factors (group 2A) did not result in the appearance of an estrogen-induced LH surge. On the other hand, prolonged estrogen priming in male castrates (group 2B) resulted in activation of the positive feedback mechanism; a LH surge in response to the estrogen provocation occurred. The results of the present study imply that 1) contrary to an earlier suggestion, testosterone does not block or blunt the LH surge, indicating that it is probably not responsible for suppressing the LH surge in normal men; 2) testosterone can cause ovulatory failure without suppressing the LH surge in women; and 3) prolonged estrogen priming may be involved in activation of the positive feedback mechanism in humans.

Adult↗

Downward regulation of plasma LH by LHRH agonist, leuprolide acetate, resulting in inhibited renal growth and function in the castrated male rat.

We previously reported that ovine and porcine luteinizing hormone (LH) stimulated kidney growth in castrated hypophysectomized rats. Our present study focuses on the physiological role of the renotropic activity of LH isoforms. Plasma LH levels were decreased to 10% of that of castrated control rats by injections of a slow-releasing LHRH agonist, leuprolide acetate, from microcapsules. Compared to controls, which were injected with microcapsules only, the kidney weight in leuprolide-treated castrated rats decreased 12%. Renal protein and DNA contents decreased significantly. Body, liver and spleen weights were not changed by the treatment, however. This effect on the kidney was not observed in castrated hypophysectomized rats, suggesting that leuprolide affected the kidneys indirectly, rather than directly, by suppressing LH secretion. In leuprolide-treated castrated rats, urinary fractional excretion of sodium (FENa) increased, indicating suppressed renal function at the proximal tubules. We concluded that the secretion of renotropically active LH isoforms was regulated at least partially by LHRH and played a physiological role in growth and the function of the proximal tubules.

Animals↗

Plasma vasopressin and oxytocin levels in intact goats and in castrated goats given testosterone.

Oxytocin, vasopressin, cortisol and testosterone levels in the plasma were measured by radioimmunoassay in intact male goats as well as in prepubertally castrated goats injected daily, for 2 weeks, with oil vehicle and then, for 4 weeks, with testosterone propionate in oil to study the influence of gonadal steroids on posterior pituitary hormones. Packed cell volume, plasma osmolality and sodium concentration were also measured in all blood samples. Plasma levels of oxytocin, vasopressin and cortisol were similar in the intact and oil-injected castrated goats. Testosterone treatment significantly increased plasma levels of oxytocin (P less than 0.01) in castrated goats but the increased levels were similar to those seen in the intact goats at the same time of year. Plasma levels of cortisol and vasopressin were unaffected by testosterone propionate treatment, whereas packed cell volume was significantly decreased (P less than 0.01). Testosterone treatment of castrated male goats appears not to have any action on pituitary hormones and oxytocin increases in the spring in both intact and castrated male goats.

Animals↗

Long-term castration decreases the androgen but not the estrogen nuclear pituitary receptors in the ram.

The effect of castration on pituitary androgen and estrogen nuclear receptors was examined in eleven 3-year-old 'Préalpes du Sud' rams which were either intact (N = 5) or surgically castrated (N = 6) 11 months before. To avoid a receptor loss, pituitaries were frozen less than 1 min after slaughter. Specific androgen and estrogen bindings were measured upon extracts from purified nuclei. Neither the concentration of estrogen receptors (30.5 +/- 3.0 vs 28.1 +/- 3.0 fmol/mg protein) nor the affinity constant (ka = 1.21 X 10(9) mol-1 in both groups) differed between intact and castrated rams. Conversely, androgen receptor concentrations differed markedly between groups and were found significantly higher in intact rams (11.3 +/- 1.1 fmol/mg protein) than in castrated rams where nuclear receptors were undetectable in 5 out of 6 animals. This result keeps open the possibility that the decrease of LH sensitivity to testosterone negative feedback observed after long-term castration, could be related to the quasi absence of nuclear receptors, at least at the pituitary level.

Animals↗

Castration-induced hyperactivity of seminal vesicle in the catfish Clarias batrachus: a case of paradox and blockade by antiandrogen (cyproterone acetate) treatment.

Castration of the catfish Clarias batrachus in late preparatory-early prespawning phase (April-May) caused time-dependent stimulatory effect on morphology, weight, and in the concentrations of biochemical correlates, such as total proteins, fructose, hexosamines and sialic acid in the seminal vesicle (SV). The peak changes were noticed on week 4 of castration. The hyperactivity was related to augmented production of testosterone by the SV of castrates with the levels significantly high from week 3 onwards. As a result, serum testosterone level fluctuated with a significant decrease in the first and fifth weeks, a significant increase in the third week, and no significant difference in the second and fourth weeks. Serum E2 level decreased significantly throughout. Cyproterone acetate treatment (CA; 1 mg/fish daily for 21 days) from the second day of castration decreased the size and weight of the SV and the concentrations of total proteins, hexosamines, fructose and sialic acid. The antiandrogen treatment did not alter serum testosterone level but the E2 level was significantly decreased. It is concluded that the hypersecretory activity of the SV in castrates is a sequel to local synthesis and action of testosterone and the effect could be prevented by CA by blocking androgen actions.

Androgen Antagonists↗

Loss of penile erectile response to intracavernous injection of acetylcholine in castrated dog.

Castrated dogs showed a slightly weak erectile response to cavernous nerve stimulation compared with normal dogs. Intracavernous injection of acetylcholine induced a dose-dependent increase of intracavernous pressure in normal dogs, however, there was almost no penile response in castrated dogs. Histological study showed a significant higher ratio of collagen tissue/smooth muscle in the corpus cavernosum of castrated dogs, but no difference in acetylcholine-esterase staining and the number of endothelial cells lining the sinusoidal spaces between castrated and normal dogs. These results indicate that the corpus cavernosum of castrated dogs nearly lost responsiveness to acetylcholine probably due to a high ratio of collagen/smooth muscle fibers and that there are different mechanisms for erectile response to nerve stimulation and intracavernous injection of acetylcholine.

Acetylcholine↗

Differential responses of hypothalamic LHRH-I and -II to castration and gonadal steroid or tamoxifen treatment in cockerels.

Changes in the hypothalamic contents of LHRH-I and LHRH-II were determined in intact and castrated cockerels injected i.m. with gonadal steroids or tamoxifen. An increase in the plasma concentration of LH after castration was accompained by a significant increase in the content of LHRH-I in the posterior hypothalamus (including the mediobasal hypothalamus and median eminence) which was reversed by oestradiol benzoate given on days 14 and 15 after castration. Under similar circumstances, testosterone propionate did not modify the hypothalamic content of LHRH-I, even though both steroids reduced the plasma concentrations of LH to levels below those of intact cockerels. Treatment of intact cockerels with oestradiol benzoate significantly increased the content of LHRH-I in the posterior hypothalamus, whilst testosterone propionate was again without effect. Tamoxifen significantly raised the plasma concentration of LH in intact cockerels and partially antagonized the suppressive effect of oestradiol benzoate and testosterone on LH secretion in castrated cockerels. However, an anti-oestrogenic effect of tamoxifen on the hypothalamic content of LHRH-I was not demonstrated. There was no evidence of any changes in the hypothalamic content of LHRH-II after castration, with or without gonadal steroid replacement. A change in the hypothalamic content of LHRH-I in response to manipulation of the steroid environment would imply an involvement of this peptide in the mechanism by which gonadal steroids regulate the release of LH. The absence of changes in the hypothalamic content of LHRH-II in the same circumstances suggest that it is not directly involved in the control of LH secretion by the gonadal steroid negative feedback loop.

Animals↗

Effect of intratesticular injection of lidocaine on cardiovascular responses to castration in isoflurane-anesthetized stallions.

OBJECTIVE: To evaluate the effect of intratesticular administration of lidocaine on cardiovascular responses and cremaster muscle tension during castration of isoflurane-anesthetized stallions. ANIMALS: 28 healthy stallions (mean +/- SD age, 4.2 +/- 2.8 years) with no testicular abnormalities that were scheduled for castration. PROCEDURE: Each horse was given acepromazine (20 microg/kg, IM), romifidine (50 microg/kg, IV), and butorphanol (20 microg/kg, IV). Anesthesia was induced with ketamine (2.5 mg/kg, IV) and midazolam (50 microg/kg, IV) and maintained with isoflurane (1.7% end-tidal concentration). After 10 minutes at a stable anesthetic plane, a needle was placed in each testicle and either no fluid or 15 mL of 2% lidocaine was injected; 10 minutes after needle placement, surgery was commenced. Pulse rate and arterial blood pressures were measured invasively at intervals from 5 minutes prior to castration (baseline) until 5 minutes after the left spermatic cord was clamped. The surgeon subjectively scored the degree of cremaster muscle tension. In 2 horses, lidocaine labeled with radioactive carbon (C(14)) was used and testicular autoradiograms were obtained. RESULTS: Compared with baseline values, castration significantly increased blood pressure measurements; intratesticular injection of lidocaine decreased this blood pressure response and cremaster muscle tension. In 2 horses, autoradiography revealed diffuse distribution of lidocaine into the spermatic cord but poor distribution into the cremaster muscle. CONCLUSIONS AND CLINICAL RELEVANCE: In isoflurane-anesthetized stallions, intratesticular injection of lidocaine prior to castration appeared to decrease intraoperative blood pressure responses and cremaster muscle tension and may be a beneficial supplement to isoflurane anesthesia.

Anesthetics, Local↗

Diagnosis, surgical treatment, and performance after unilateral castration in breeding bulls: 21 cases (1989-1999).

OBJECTIVE: To evaluate signalment, surgical treatment, postoperative complications, and future breeding success or semen production in a group of bulls with naturally occurring disease of the scrotum or testis. STUDY DESIGN: Retrospective study. ANIMALS: 21 bulls that underwent unilateral castration after evaluation for scrotal swelling. PROCEDURE: A computer-assisted search of medical records at 2 veterinary teaching hospitals was performed. Historical, diagnostic, surgical, and follow-up data were collected and analyzed for those bulls with scrotal swelling that underwent unilateral castration. RESULTS: Four of 5 pasture breeding bulls and 9 of 10 semen collection-center bulls successfully bred cows or produced viable semen within 6 months of surgery. Fourteen of 21 surgical procedures were performed after induction of general anesthesia. Sixty-six percent of procedures were performed as open castrations. Seventy-one percent of bulls developed postoperative complications, most of which were mild swellings. Unilateral castration returned 13 of 15 bulls with unilateral disease of the scrotum or testis to productive service by 6 months after surgery. CONCLUSION AND CLINICAL RELEVANCE: Unilateral castration is an effective treatment for unilateral disease of the,scrotum or testis in bulls, allowing return to reproductive function.

Animals↗

Effects of suppressing cortisol following castration of bull calves on adrenocorticotropic hormone, in vitro interferon-gamma production, leukocytes, acute-phase proteins, growth, and feed intake.

The objective was to determine the effects of reducing the plasma cortisol rise in calves following castration on plasma ACTH concentrations, keyhole limpet hemocyanin (KLH)- and concanavalin A (Con A)-induced in vitro interferon (IFN)-gamma production, white blood cell (WBC) numbers, neutrophil:lymphocyte (N:L) ratio, plasma haptoglobin and fibrinogen concentrations, ADG, and ADFI. Forty 5-mo-old Friesian bull calves (169 +/- 1.7 kg) were assigned to four treatments: 1) control (CON); 2) oral metyrapone administration (MET); 3) surgical castration at 0 h on d 0 (SURG); and 4) oral metyrapone administration and surgical castration (MET+SURG). Cortisol, ACTH, IFN-gamma production, haptoglobin, fibrinogen, ADFI, and ADG were not different between CON and MET animals. The MET+SURG calves had lower (P < .001) peak and mean cortisol during .25 to 1.5 h than SURG animals, but area under the cortisol vs time curve from 0 to 12 h did not differ (P > .39) between SURG and MET+SURG calves. Peak ACTH concentrations and area under the ACTH vs time curve from 0 to 6 h were greater (P < .05) for MET+SURG than for SURG calves. There were no differences between MET+SURG and SURG animals in IFN-gamma production, WBC numbers, and ADFI. On d 1, MET+SURG and SURG animals had lower (P < .01) KLH- and Con A-induced IFN-gamma production and higher (P < .05) neutrophil numbers and N:L ratio compared with CON animals. Plasma haptoglobin on d 1 and 3 and fibrinogen concentrations on d 3 and 7 were elevated (P < .05) for MET+SURG and SURG compared with CON animals, whereas SURG animals had greater (P < .05) haptoglobin and fibrinogen concentrations than MET+SURG animals on d 7. The ADG of SURG calves was lower (P < .05) than that of MET+SURG calves during d 0 to 7. Metyrapone treatment partially suppressed cortisol and increased ACTH in castrated calves but did not alter the castration-induced suppression of IFN-gamma and increases in neutrophil numbers and the N:L ratio.

Acute-Phase Proteins↗

Effects of ketoprofen alone or in combination with local anesthesia during the castration of bull calves on plasma cortisol, immunological, and inflammatory responses.

To determine the effects of the anti-inflammatory ketoprofen, alone or with local anesthesia (LA) during castration on cortisol, immune, and acute phase responses, 40 Friesian calves (215 +/- 3.5 kg) were assigned as follows: 1) control, 2) surgical castration (SURG), 3) SURG following ketoprofen (3 mg/kg BW i.v.; SURG + K), 4) SURG following LA (9 mL of 2% lidocaine hydrochloride to each testis; SURG + LA), or 5) SURG following LA and K (SURG + LA + K). Total cortisol response was greater (P < 0.05) in SURG, SURG + LA, and SURG + K + LA calves than in control calves and was not different between control and SURG + K calves. The interval to peak cortisol was longer (P < 0.05) for SURG + K + LA calves than for either SURG or SURG + K calves. On d 3, KLH-induced interferon-gamma production was lower (P < 0.05) in SURG calves than in control calves, whereas concanavalin A-induced interferon-gamma production was lower (P < 0.05) in all castration groups than in control. On d 1 after surgery, fibrinogen was higher (P < 0.05) in SURG and SURG + LA calves than in control calves, whereas SURG + LA + K calves had lower (P < 0.05) fibrinogen than did SURG calves. Haptoglobin was higher (P < 0.05) in SURG calves on d 1, 3, and 7 than in control calves. On d 1 after surgery, SURG + K and SURG + LA + K calves had lower (P < 0.05) haptoglobin concentrations than SURG calves, whereas SURG + K calves had lower (P < 0.05) levels than SURG calves on d 3. In conclusion, surgical castration induced a significant elevation in cortisol secretion; the rise in cortisol was reduced to control levels by the administration of ketoprofen but not local anaesthetic. Thus, systemic analgesia using ketoprofen is more effective than local anesthesia during castration to alleviate the associated stress response.

Acute-Phase Proteins↗

Castration lowers and testosterone restores blood pressure in several rat strains on high sodium diets.

The objective of this study was to determine the effect of a high sodium diet and prepubertal castration (5-6 weeks) and androgen replacement therapy on blood pressure in male normotensive, borderline hypertensive and hypertensive rats on a high sodium diet between 9-22 weeks of age. The strains used were: Wistar Kyoto-(WKY), spontaneously hypertensive rat-(SHR), and borderline hypertensive rat-(BHR). Castration significantly reduced blood pressure (20-30 mmHg) and testosterone replacement in castrated males restored blood pressure in all strains. Plasma norepinephrine (NE) increased with castration in the WKY and SHR strains but decreased in the BHR. However, there was a significant elevation in all strains between the midpoint and endpoint NE values. The high sodium diet did not prevent the blood pressure lowering effect of castration.

Animals↗

Effects of castration on thymocyte development in two different models of thymic involution.

Age-associated thymic involution is accompanied by decreased thymic output. This adversely affects general immune competence and T cell recovery following cytoreductive treatments such as chemotherapy. A causal link between increasing sex steroids and age-related thymic atrophy is well established. Although castration has been demonstrated to regenerate the atrophied thymus, little is known about how this is initiated or the kinetics of thymocyte regeneration. The present study shows that although castration impacts globally across thymocyte development in middle-aged mice, the regenerative effects are initiated in the immature triple-negative compartment and early T lineage progenitors (ETP). Specifically, there was a reduction in number of ETP with age, which was restored following castration. There was, however, no change in ETP reconstitution potential in ETP at this age or following castration. Furthermore, in a chemotherapy-induced model of thymic involution, we demonstrate castration enhances intrathymic proliferation and promotes differentiation through the triple-negative program. Clinically, reversible sex steroid ablation is achieved hormonally, and thus presents a means of ameliorating immune inadequacies, for example, following chemotherapy for bone marrow transplantation. By improving our understanding of the kinetics of thymic recovery, this study will allow more appropriate timing of therapy to achieve maximal reconstitution, especially in the elderly.

Aging↗

[Review of adjuvant breast cancer therapy in non-menopausal women including early results of medical castration with LH-RH analogs].

Three treatment modalities have successively dominated adjuvant therapy of breast cancer in non-menopausal women, namely, castration, chemotherapy and tamoxifen administration. The benefits afforded by each of these modalities seem similar when the treatments are compared indirectly by meta-analysis. Once the anti-tumour action of LH-RH analogues and their reversible action on ovarian function had been established, these analogues were used instead of surgical castration in direct comparisons of the three treatment modalities. Most of the patients in these trials had estrogen and/or progesterone receptor positive tumours. According to the current state-of-the-art and whilst awaiting the final results of ongoing trials, we can conclude that: The survival of surgically castrated patients is the same as that of patients who have received CMF-type chemotherapy. The survival of patients on tamoxifen is the same as that of patients who have received CMF-type chemotherapy if tamoxifen is administered for 5 years. It is lower if tamoxifen is given for only 2 years. In 2 out of 3 trials, patients receiving the combined treatment castration plus tamoxifen had improved recurrence-free survival rates compared to patients on chemotherapy (regardless of whether an anthracyclin was included or not. It is too early to comment on overall survival. Combining castration and chemotherapy seems to be advantageous in patients less than forty and in those in whom chemotherapy has not induced amenorrhea. Combining tamoxifen and chemotherapy markedly decreases the risks of disease recurrence and of death but the high standard deviations recorded mean that this statement has to be tempered. Finally, an arrest of ovarian function by LH-RH analogues that is only temporary apparently does not adversely impinge upon results. This has, however, to be proved in an ad hoc trial and the optimum duration of analogue administration has to be established.

Antineoplastic Agents, Hormonal↗

beta-glucuronidase activity is elevated in the rat harderian gland after bromocriptine but decreased after cyproterone acetate treatment or castration. Postcastrational effects on the ultrastructure of the gland.

OBJECTIVE: To investigate the effect of castration, bromocriptine and cyproterone acetate treatment on lysosomal hydrolytic enzymes and ultrastructure of the rat Harderian gland. METHODS: Groups of rats were subjected to the treatment by bromocriptine, cyproterone acetate and castration. Harderian glands were dissected from each experimental animal cut into small pieces and immediately stored in liquid nitrogen. Only those from castrated animals were fixed in Karnovsky fluid and processed for electron microscopy. The activity of acid phosphatase and ss-glucuronidase were estimated by kits from Sigma (St. Louis, Mo, USA). Semi-thin sections were stained with 1-% toluidine blue and ultra-thin sections with uranyl acetate and counter-stained with lead citrate. The examination was performed by Joel JEM-1200 EX II electron microscope. RESULTS: The treatment of male rats with the prolactin release inhibitor bromocriptine induced a significant increase in ss-glucuronidase activity. On the contrary, such activity was significantly decreased after treatment with the testosterone receptor antagonist cyproterone acetate as well as after castration. However, these treatments did not alter the activity of lysosomal acid phosphatase. Castration induced dramatic changes mostly in type A cells such as the appearance of lipid vacuoles with irregular forms and the predomination of smooth endoplasmic reticulum (SER) throughout the cytoplasm. The most dramatic postcastrational change was the degeneration of the mitochondria. These changes in type A cells might be due to the uneven distribution of the testosterone receptors in the rat Harderian gland, which are more numerous in type A cells. CONCLUSIONS: the gland physiology is responsive to alteration in circulating prolactin and testosterone levels.

Acid Phosphatase↗