Weight In Ovariectomized Rats Research Articles

Structure-Activity Relationships of Triphenylethylene Derivatives and Their Evaluation as Anticancer and Antiviral Agents.

Tamoxifen (TAM) is a selective estrogen receptor modulator (SERM) that is used in the treatment of breast cancer, yet with the risk of developing uterine cancer. A perfect SERM would act as an estrogen activator on bones, the cardiovascular system, and the central nervous system while providing neutral or estrogen blocking effects on the breast and the uterus. Herein, we report on the design, synthesis, and evaluation of new rigid and flexible TAM analogues. Mainly, a chloro substituent is introduced at the para position of the TAM ring C blocking the CYP2D6 hydroxylation site. Most compounds showed estrogenic activity higher than TAM using the yeast estrogen screen assays, indicating the determinant role of the chloro substituent upon functional activity. Despite being estrogenic, compound 2B showed potent antiproliferative activity in the NCI 60 cell lines with mean GI50 = 3.67 μM, GI50 = 1.05 μM on MCF-7 cell lines, and GI50 = 1.30 μM on MDA-MB-231. The estrogenic activity of compound 2B was further confirmed by stimulating alkaline phosphatase in Ishikawa cells, and it showed no increase in relative uterine wet weight in ovariectomized rats. Compound 2F showed EC90 = 0.31 μg/mL and SI90 = 60 against Ebola virus; this is 200-fold more potent than the positive control favipiravir. This is the first time to report estrogenic triphenylethylenes as anti-EBOV agents. The anti-EBOV activity reported is a function of the substitution pattern of the scaffold rather than the functional activity. Moreover, compound 3D showed excellent PO pharmacokinetic properties in mice. In conclusion, for this class of TAM-like compounds, the blockage of the p-position of ring C is decisive for the functional activity; meanwhile, the triarylethylene substitution pattern is detrimental for the antiviral activity.

Effect of 50-Hertz Sinusoidal Vibration on the Uterus in Ovariectomy-Induced Osteoporotic Rats.

Objective To evaluate the influence of sinusoidal vibration (50-Hertz) stimulation on the uterus of osteoporotic rats. Methods We constructed an osteoporosis rat model by ovariectomy (OVX). 36 3-month-old Sprague Dawley rats were randomly divided into the control group, vibrating group, sham operation group, sham operation vibrating group, OVX group, and OVX vibrating group (n = 6 per group). Rats started to vibrate one week after the operation: one 10 minutes 50-Hertz sinusoidal vibration per day, except for Saturday and Sunday. In the second, 8, and 12 week after vibration stimulation, rats were sacrificed in batches. And then, the uteruses were taken out to measure the wet weight and calculate uterus relative wet weight. Results Compared with the control group, OVA induced a significant increase in wet weight and relative wet weight in rat uterus. The vibration was to the uterus wet weight and the uterus relative wet weight in ovariectomized rats and at the same time had no significant effect, but the 12-week prolonged vibration can significantly reduce the uterus wet weight and the uterus relative wet weight in ovariectomized rats than 2 weeks. Conclusions The uterus wet weight and the uterus relative wet weight were increased in the OVA-induced osteoporosis rats. The 50-Hertz sinusoidal vibration had no significant effect on the uterus wet weight and the uterus relative wet weight in the ovariectomized rats at the same time, but 12 weeks of vibration can significantly reduce the uterine wet weight and uterine relative wet weight of ovariectomized rats. And the uterus relative wet weight can be used as a new indicator of stimulating the uterus.

Uterus Weight of Ovariectomized Rats Given Cocoa Powder and Extract

Estrogen is a hormone that influences the growth of uterus. Ovariectomized rats lose their ovaries as the main source of estrogen so their uterus tends to shrink. The objective of thise study was to evaluate the potency of cocoa as an estrogen substitute, which was done through an experiment that tested the influence of cocoa products on the uterus weight of ovariectomized rats. There were five treatments and in each of the treatments, six wistar rats were given the substance for three days. Four groups of treatment consisted of ovariectomized rats given cocoa extract (875.2 ppm of polyphenols, 1 g/kg of body weight, BW), cocoa powder (369.8 ppm of polyphenols, 1 g/kg BW), olive oil (10 mL/kg BW), or estradiol valerate (1 mg/kg BW). The fifth group consisted of intact (non-ovariectomized) rats given 10 mL/kg BW of water. The finding showed that the ovariectomized rats given olive oil had lower uterus weight than that of intact rats, while the ovariectomized rats given estradiol valerate had higher uterus weight compared to the intact and olive oil groups. The ovariectomized rats given cocoa powder and extract had higher uterus weight compared to those given only olive oil; although they were not significantly different. Correlation between the body weight and uterus weight varied across treatment groups. The rats given olive oil showed significant, positive correlation, while the intact rats showed moderate, positive correlation. The rats given estradiol valerate and cocoa powder showed non-significant correlation. Since the polyphenol content in cocoa powder was at lower concentration than that in the cocoa extract, it was predicted that cocoa polyphenols are more potential in the lower concentration. This study concludes that even though consumption of cocoa powder and extract did not significantly induced uterus growth, cocoa is still considered having estrogenic activity by lowering the correlation between the body weight and uterus weight in ovariectomized rats.

Therapeutic effect of aqueous extract from Ecliptae herba on bone metabolism of ovariectomized rats

Ecliptae herba (EH) has long been used in China to strengthen bones. Accumulating evidence indicates that EH may have antiosteoporotic effects. The aim of this study was to evaluate the effects of aqueous EH extract (EHE) on rats that had osteoporosis-like features induced by ovariectomy, using aqueous Fructus Ligustri Lucidi extract as positive control agent. Three-month-old female rats that underwent ovariectomy were treated with EHE (1.4 g/kg per day). After 12 weeks, bone mineral density and bone histomorphometric indices of tibiae were measured. Protein and messenger RNA expressions of osteoprotegerin and receptor activator of nuclear factor κ-B ligand (RANKL) in tibiae were evaluated by immunohistochemistry and in situ hybridization. In addition, serum concentrations of osteocalcin, interleukin-1β, interleukin-6 (IL-6), calcitonin (CT), and parathyroid hormone were determined by enzyme-linked immunosorbent assay. EHE treatment prevented body weight gain and loss of uterine wet weight in ovariectomized rats. It remarkably increased bone mass in ovariectomized rats compared with ovariectomized controls. EHE treatment significantly down-regulated RANKL expression in tibiae from ovariectomized rats compared with controls; however, it had no significant effect on osteoprotegerin expression. In addition, EHE treatment significantly reduced serum IL-6 levels and remarkably increased CT levels but had no effect on parathyroid hormone. EHE increases bone mass in ovariectomized rats by inhibiting bone loss: down-regulated RANKL expression in tibiae and IL-6 level in serum, and up-regulated CT level in serum. This suggests that EHE may be developed as an alternative therapeutic agent for osteoporosis induced by postmenopause.

Effects of the novel orally active antiestrogen TZE-5323 on experimental endometriosis.

Danazol and gonadotropin-releasing hormone agonists which are used as therapeutic drugs for endometriosis, develop adverse reactions in association with their long-term use. The efficacy of anti-estrogens for endometriosis, an estrogen-dependent disorder, has not been demonstrated. A novel, orally active anti-estrogen, TZE-5323 ((2-cyclohexy-6-hydroxybenzo[b]thien-3-yl)[4-[2-(1- piperidinyl)ethoxy]phenyl] methanone hydrochloride, CAS 150797-71-0; free salt formula) was developed. TZE-5323 showed strong affinity for human estrogen receptor alpha (hER alpha) and beta (hER beta), and dose-dependently inhibited estradiol-stimulated transcriptional activation via hER alpha and hER beta. Furthermore, TZE-5323 dose-dependently reduced estrogen-increased uterine weight in ovariectomized rats. Tamoxifen showed agonistic activity on hER alpha, while TZE-5323 did not show such activity. In the experimental endometriosis model in rats in which endometrial tissue is autotransplanted into the renal subcapsular space, TZE-5323 dose-dependently reduced the volume of the endometrial implant as did danazol and leuprorelin acetate. Furthermore, the long-term administration of TZE-5323 neither showed a decrease in bone mineral density nor did it affect serum estradiol concentrations in intact rats. Therefore, TZE-5323 suggested its potential as a novel therapeutic drug for endometriosis which is effective also in long-term use.

Low-magnitude whole body vibration does not affect bone mass but does affect weight in ovariectomized rats

Mechanical loading has stimulating effects on bone architecture, which can potentially be used as a therapy for osteoporosis. We investigated the skeletal changes in the tibia of ovariectomized rats during treatment with whole body vibration (WBV). Different low-magnitude WBV treatment protocols were tested in a pilot experiment using ovariectomized rats with loading schemes of 2 × 8min/day, 5days/week (n=2 rats per protocol). Bone volume and architecture were evaluated during a 10week follow-up using in-vivo microcomputed tomography scanning. The loading protocol in which a 45Hz sine wave was applied at 2Hz with an acceleration of 0.5g showed an anabolic effect on bone and was therefore further analyzed in two groups of animals (n=6 each group) with WBV starting directly after or 3weeks after ovariectomy and compared to a control (non-WBV) group at 0, 3, 6 and 10weeks' follow-up. In the follow-up experiment the WBV stimulus did not significantly affect trabecular volume fraction or cortical bone volume in any of the treatment groups during the 10week follow-up. WBV did reduce weight gain that was induced as a consequence of ovariectomy. We could not demonstrate any significant effects of WBV on bone loss as a consequence of ovariectomy in rats; however, the weight gain that normally results after ovariectomy was partly prevented. Treatment with WBV was not able to prevent bone loss during induced osteoporosis.

Effect of Soybean Seeds On Some Physiological And Biochemical Parameters In Ovariectomized Rats

The present study was conducted to investigate the effect of soybean seeds (15g /kg diet) for one month on blood picture, total cholesterol(T-C), high density lipoproteins(HDL), low density lipoproteins(LDL), triglycerides (TG), glucose, alanine aminotrasferase (ALT) and aspartate aminotrasferase (AST) enzymes activity, glutathione(GSH), total proteins, albumin, globulin and body weight in ovariectomized rats(OVX). The rats randomly divided in to three groups (6 rats / group). The first group served as a control, the second group rats at one month aged bilateral ovariectomy were done, and the third group rats at one month aged bilateral ovariectomy were done, and after two months the rats treated with soybean seeds for one month. The result showed that a significant decrease in the total leucocytes count, HDL, GSH and globulin in ovariectomized group compared with control group, with no significant differences in PCV, Hb, differential leucocytes count(DLC), total proteins, albumin and body weight. The current study demonstrated that a significant increase in T-C, LDL, ALT and glucose in ovariectomized rats group compared with control group. Administration of soybean seeds caused a significant increase in total leucocytes count, globulin and body weight compared with ovariectomized group, and a significant decrease in T-C, TG, LDL and glucose compared with ovariectomized group. It could be concluded from this study that administration of soya beans seeds have a beneficial improving of some physiological and biochemical parameters in ovarictomized rats.

Simvastatin induces estrogen receptor-alpha expression in bone, restores bone loss, and decreases ERα expression and uterine wet weight in ovariectomized rats

We previously reported that simvastatin induces estrogen receptor-alpha (ERα) in murine bone marrow stromal cells in vitro. In this study, we investigated the effect of simvastatin on ERα expression in bone and uterus in ovariectomized (OVX) rats and evaluated bone mass, bone strength, and uterine wet weight. Three-month-old Sprague-Dawley female rats received OVX or sham operation. Six weeks later, the rats were treated orally with simvastatin (5 or 10mg/kg/day), or intraperitoneally with 17-β-estradiol (E(2)) or a combination of simvastatin and E(2) for 6weeks. Uterine wet weight, bone mineral density (BMD) of lumbar vertebrae, biomechanics of lumbar vertebrae, and induction of ERα expression in the bone and uterus were analyzed. The 6-week simvastatin treatment improved lumbar vertebral BMD and boosted biomechanical performance of the vertebral body compared to the OVX control, suggesting that simvastatin can treat osteoporosis caused by estrogen deficiency. More interestingly, simvastatin could increase ERα expression and synergy with estradiol in bone while antagonizing estradiol in the uterus, along with uterus atrophy and uterine wet weight decreases. In conclusion, these data suggest that simvastatin exert opposing modulatory effects on ERα expression on bone and uterus in ovariectomized rats, inducing ERα expression and synergy with estrogen to perform anabolic effects on the bones while decreasing E2 efficacy and uterine wet weight. This finding may be helpful to explain the mechanism of statin treatment in osteoporosis caused by estrogen deficiency.

87 Effects of Horse Bone Extracts on Induced Postmenopausal Osteoporosis in Rats

− Osteoporosis is a metabolic bone disease associated with an imbalance of bone remodeling. Osteoporosis is characterized by decreased bone mass and increased bone fractures. In this study, we investigated the effects of horse bone extracts (HBEs) in vivo. Horse bone was extracted with 80% alcohol (HBE-A) at 100 o C or water (HBE-W) at 120 o C. Animal model of postmenopausal osteoporosis was used, in which osteoporosis was induced by ovariectomy of female S.D. rats (female rats were divided into 5 groups), and HBEs were administered to ovariectomized rats every day for 8 weeks. After 8 weeks, the rats were sacrificed and the following osteoporotic factors were measured: body weight, bone mineral density (BMD), uterine/body weight ratio, serum estradiol (E2), and serum alkaline phosphatase (ALP). The results showed that the administration of HBE-W decreased the changes of body weight in ovariectomized rats. HBE-W increased the uterine/body weight ratio and BMD. In addition, HBEs decreased the ALP. Therefore, HBEs may be used for the prevention or treatment of bone disease.

Effects of Zingerone on Fat Storage in Ovariectomized Rats

We reported that ginger prevented obesity in mice fed a high-fat diet in previous study. In this experiment, we examined the effects of zingerone, the major pungent component of ginger on fat storage in ovariectomized (Ovx) rats. Oral administration of 170 mg/kg zingerone significantly reduced body weight and the final parametrail adipose tissue weight in ovariectomized rats. Blood glucose levels after oral administration of glucose were lower in zingerone-treated Ovx-rats than in the Ovx-rats (control). Basal lipolysis in zingerone-treated Ovx-rats was increased compared with that in the Ovx-rats. Zingerone significantly increased norepinephrine-induced lipolysis associated with the translocation of hormone-sensitive lipase (HSL) from the cytosol to lipid droplets in adipocytes. These results indicate that zingerone may prevent the fat storage through increasing norepinephrine-induced lipolysis in adipocytes.

Effects of Labisia pumila var alata extracts and estrogen replacement therapy on body weight gain, adipokine levels and uterus weight in ovariectomized rats

Effects of Labisia pumila var alata extracts and estrogen replacement therapy on body weight gain, adipokine levels and uterus weight in ovariectomized rats

Gynecological efficacy and chemical investigation of Vitex agnus-castus L. fruits growing in Egypt

Flavonoid glycosides, orientin and apigenin 3, 8-di-C-glycosides in addition to, iridoid compound, aucubin were isolated from the ethanolic extract of Vitex agnus-castus fruits. Their structures were identified on the basis of the spectroscopic data. The estrogenic activity of the ethanolic extract in two dose levels 0.6 and 1.2 g kg−1 per body weight (b.w.) was studied by the vaginal smear, and uterine weight methods for normal and ovariectomized female rats. The extract induced significant increase in the uterine weight of ovariectomized rats at two dose levels comparable to that of control group. The percentages of the total average number of scores were increased significantly too. Significant increases in plasma progesterone and total estrogens levels were shown at the two dose levels when compared to that of control group. On the other side, the extract induced significant reduction in lutinizing and plasma prolactin hormones.

Estrogen replacement regimen and brain infusion of lipopolysaccharide differentially alter steroid receptor expression in the uterus and hypothalamus

The regimen of estrogen replacement can alter the consequences of estrogen therapy and stressors. To determine the long-term effects and interaction of these systems on the brain and periphery, adult female rats were infused with lipopolysaccharide (LPS) into the fourth ventricle of the brain for 4 weeks, and ovariectomized rats were administered either constant or pulsed regimens of estrogen replacement (17beta-estradiol) until sacrifice at 8 weeks. Constant, but not pulsed, estrogen replacement reduced ERalpha and increased HSP90, HSP70, and PR(B) uterine protein levels. Both estrogen regimens increased ERbeta, HSP27, and PR(A) uterine proteins. Both regimens reduced hypothalamic levels of ERalpha, but not ERbeta, HSP, or PR. No changes were observed in the hippocampus. Long-term brain infusion of LPS activated microglia and reduced body weight, but did not alter corticosterone or nitrotyrosine levels. LPS infusion into intact rats suppressed uterine weight, increased ERalpha and decreased HSP90 in the uterus. LPS did not alter uterine weight in ovariectomized rats treated with constant or pulsed estrogen. Together, these data suggest the timing of estrogen replacement and neuroinflammatory stressors can profoundly affect uterine and hypothalamic steroid receptor expression and may be important parameters to consider in the post-menopausal intervention with estrogen.

Acute activation of ERα decreases food intake, meal size, and body weight in ovariectomized rats

Estradiol exerts many of its actions by coupling with two nuclear estrogen receptor (ER) proteins, ER alpha, and ER beta. While the acute, anorexigenic effect of estradiol appears to involve such a mechanism, the relative contributions of ERalpha and ERbeta are equivocal. To address this problem, food intake was monitored in ovariectomized (OVX) rats following acute administration of a selective ER alpha agonist (4,4',4''-(4-propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol, PPT; dose range = 0-200 microg), a selective ER beta agonist (2,3-bis(4-hydroxyphenyl)-propionitrile, DPN; dose range = 0-600 microg), and a physiological (4 microg) dose of estradiol benzoate (EB). While PPT-treated rats displayed dose-dependent decreases in daily food intake and body weight, neither of these measures was influenced by any dose of DPN. In addition, DPN failed to modulate the anorexigenic effect of PPT when the two ER agonists were coadministered. Meal pattern analysis revealed that the anorexigenic effect of 75 microg PPT (a dose of PPT that produced a similar decrease in daily food intake as 4 microg EB) was mediated by a decrease in meal size, not meal number. Thus, PPT, like EB and endogenous estradiol, decreases food intake by selectively affecting the controls of meal size. The finding that acute administration of 75 microg PPT failed to induce a conditioned taste aversion suggests that the anorexigenic effect of this dose of PPT is not secondary to malaise. Taken together, our findings demonstrate that selective activation of ER alpha decreases food intake, body weight, and meal size in the ovariectomized rat.

Cyclic Estradiol Treatment Normalizes Body Weight and Test Meal Size in Ovariectomized Rats

We tested whether cyclic estradiol treatment, like continuous estradiol treatment, is sufficient to normalize meal size and body weight in ovariectomized rats. In Experiment 1, adult Long–Evans rats were ovariectomized and subcutaneously injected with 0, 0.2, or 2.0 μg estradiol benzoate (EB) in sesame oil each Tuesday and Wednesday. Oil-treated ovariectomized rats gained more weight during 4 weeks of ad lib feeding (48 ± 5 g) than intact rats (16 ± 1 g, p < 0.01). Cyclic treatment with 2.0 μg EB normalized weight gain (11 ± 2 g). During the next week, plasma samples were assayed for estradiol. Cyclic treatment with 2.0 μg EB produced excursions of plasma estradiol that appeared similar to those of intact, cycling rats: estradiol level reached 190 ± 60 pmol/L after the second EB injection before decreasing to undetectable levels (<30 pmol/L) by cycle end. In Experiment 2, test meal sizes after overnight food deprivation were measured. Cyclic treatment with 2.0 μg EB produced both tonic (i.e., at cycle onset, meal size was smaller in estradiol-treated than oil-treated rats) and phasic (i.e., meal size was smaller late in the EB-treatment cycle than early in it) decreases in meal size. Thus, a weekly cyclic regimen of estradiol treatment that produces changes in plasma estradiol concentration similar to those in intact cycling rats is sufficient to produce the body weight and meal size patterns that characterize normal hypothalamic–pituitary–gonadal function.

Effect of the high-affinity estrogen receptor ligand ICI 182,780 on the rat tibia.

We examined the specificity of the steroidal antiestrogen ICI 182,780 (ICI) on bone and reproductive tissues in adult and growing female rats. Using a 1.5-mg/kg dose (s.c.), we evaluated the effects of ICI on the bone, body weight, uterine weight, serum cholesterol, and serum estradiol in either adult and/or growing rats. ICI increased serum estradiol cholesterol in ovary-intact rats, had no effect on uterine weight in ovariectomized rats, and resulted in uterine atrophy in ovary-intact animals comparable with ovariectomy. In contrast, ICI had no effect on body weight. In bone, ICI significantly increased the rate of periosteal bone formation in long bones of growing and mature female rats. In contrast, ICI had no effect on longitudinal bone growth in rapidly growing rats. When ICI was administered to mature rats with or without ovaries, two-factor ANOVA revealed significant interaction (P < or = 0.05) between ovariectomy and ICI treatment for cancellous bone area and labeled bone perimeter. ICI increased skeletal indices of bone turnover in the cancellous bone of ovary-intact rats but reduced these indices of bone turnover in the cancellous bone of ovariectomized rats. The increase in bone turnover was associated with a reduction in cancellous bone area in the ovary-intact rats. A reduction in bone turnover was similarly associated with an increase in bone area in the ICI-treated ovariectomized rats. In summary, ICI exhibited complete estrogen antagonism in cortical and cancellous bone, partial agonism in cancellous bone, and no activity on tibial longitudinal growth rate of growing ovary-intact rats. The effects in adult rats were influenced by circulating levels of estradiol. ICI had no activity on body weight and complete antagonism on uterine weight. These results demonstrate that a ligand with high binding affinity to the estrogen receptor(s) can elicit an array of estrogen-mediated regulation of bone metabolism.

Changes in ingestive behaviors and body weight following intracranial application of 17 α-estradiol

The present experiment was conducted to determine if central implants of 17 α-estradiol could influence food intake, water intake, and body weight in ovariectomized rats. A total of fifteen animals were fitted with bilateral guide cannulae in the paraventricular nucleus (PVN) and stimulated unilaterally with cholesterol and 17 α-estradiol in each side of the brain. Compared with cholesterol treatment, 17 α-estradiol implants in the PVN significantly lowered food intake and body weight but did not affect water intake. These findings indicate that the PVN is a brain region responsive to the effects of 17 α-estradiol on feeding behavior, and support the hypothesis that the effects of estrogens on ingestive and reproductive behaviors are organized separately within the brain.

Dexfenfluramine: action with estradiol on food intake and body weight in ovariectomized rats

Previous work from this laboratory has shown that chronic administration of dexfenfluramine (DF) caused substantial weight loss in rats that were overweight 3-4 mo after ovariectomy (OVX), but not in OVX rats that were of normal weight, as a result of estrogen replacement. The present study was conducted to determine whether the enhanced weight loss in the former group is because of either overweight per se or an inhibitory effect of estrogen on DF. Starting either 0, 6, or 14 wk after OVX, when weight gain was zero, moderate, or near maximal, respectively, rats received a 12-day regimen of either estradiol or the oil vehicle and either DF (3 mg.kg-1.day-1 by osmotic minipump) or no drug. DF had no effect on either food intake or weight gain of groups treated during 0-2 wk after OVX but had significant anorectic and weight loss actions in groups treated 6-8 and 14-16 wk after OVX. Estrogen had a similar effect at all three times and in the 14-wk group produced an effect that was additive with that of DF. Measures of plasma glucose and triglycerides and adipose tissue lipoprotein lipase activity did not correlate with the effectiveness of the drug to promote weight loss.

Interaction of 16 alpha-[125I]iodo-estradiol with estrogen receptor and other steroid-binding proteins.

This communication describes the synthesis of 16 alpha-[125I]iodo-estradiol (125I-E2) with specific activities greater than 1000 Ci/mmol (1 Ci = 3.7 x 10(10) becquerels). We show that it binds to the same specific estrogen receptor sites as does [3H]estradiol and that it does so with an affinity that is indistinguishable from that for the latter steroid. This is true for receptor obtained from calf uterus and from a pool of human mammary carcinomas. There is no significant binding of 125 I-E2 to the testosterone-estradiol-binding globulin of human plasma. 125I-E2 also binds tightly to anti-estradiol antibodies raised against estradiol derivatized at carbons 3, 6, and 17. Finally, we show that unlabeled I-E2 is an active estrogen in vivo as demonstrated by its ability to increase uterine weight in ovariectomized rats.

The effects of estradiol on food intake and weight in ovariectomized rats with amygdaloid lesions

The effects of estradiol on increased food intake and body weight gain induced by ovariectomy, were examined in rats with lesions of the cortical and medial amygdaloid nuclei. Lesioned animals displayed typical ovariectomy-induced hyperphagia and weight gain. Estradiol injections were effective in maintaining normal food intake and weight gain in ovariectomized lesioned animals.