Parathyroid hormone-related peptide may increase mammary blood flow.

Amino-terminal fragments of PTHrP were previously shown to increase regional blood flow in laboratory animals. Since PTHrP is produced in the lactating mammary gland and associated nutrient vessels, we examined the effects of peptide fragments of PTHrP on the hemodynamics of the mammary gland of dried sheep. The left arterial mammary blood flow measured using ultrasonic flow probes in four dried Lacaune ewes was 233 +/- 11 ml/minute. It was significantly increased when synthetic human PTHrP-(1-34) or (1-86) fragments were injected into the mammary artery. The effect was dose dependent for PTHrP-(1-34), varying between 0.0075 and 0.3 nmol/kg body weight. PTHrP-(140-173) fragment lacked any vasorelaxant activity. Synthetic human endothelin (ET1) decreased arterial blood flow in a dose-dependent manner. This decrease was inhibited by PTHrP-(1-34), and this inhibition was PTHrP dose related. When ET1 (10 pmol/kg body weight) was injected together with PTHrP-(1-86) (100 pmol/kg body weight), only a significant increase in mammary blood flow was observed. Thus, PTHrP produced by the lactating mammary gland may be involved in the regulation of mammary blood flow.

Ontogenesis of calcitonin mRNA in the rabbit.

Since plasma calcium levels are higher in the fetus than in the mother at the end of gestation, it has been suggested that calcitonin (CT) biosynthesis would be very active in the fetus. This hypothesis was tested in rabbit fetuses and newborns by measuring the amount of CT mRNAs found in the thyroid glands and the thyroidal CT stores. Dot-blot and Northern hybridizations with a specific CT cDNA probe (a BglII-NsiI fragment of the human CT cDNA) were used to determine the CT mRNA level. In fetuses, newborns, and mothers, only one molecular species of mRNA around 1 kb was detected by Northern hybridization with the specific CT cDNA probe. By dot-blot, CT mRNAs could be detected at 20 days of gestation on pooled fetal thyroid glands as a weak positive signal. The amount of CT mRNAs increased on day 24; at this stage they were also observed by Northern hybridization. During the last 6 days of gestation a 3-fold increase in CT mRNAs occurred in rabbit fetuses; concomitantly a 5-fold rise in the total thyroidal CT content was observed. Fetal plasma concentrations of both CT and calcium increased slightly between 24 and 30 days of gestation. After birth, the CT mRNA level was 10-fold increased between 2 and 30 days; these changes were not reflected in the plasma CT level but were probably accounted for by a rise in the number of C cells of the thyroid gland.

Rutin inhibits ovariectomy-induced osteopenia in rats.

Several studies suggest that polyphenols might exert a protective effect against osteopenia. The present experiment was conducted to observe the effects of rutin (quercetin-3-O-glucose rhamnose) on bone metabolism in ovariectomized (OVX) rats. Thirty 3-month-old Wistar rats were used. Twenty were OVX while the 10 controls were sham-operated (SH). Among the 20 OVX, for 90 days after surgery 10 were fed the same synthetic diet as the SH or OVX ones, but 0. 25% rutin (OVX + R) was added. At necropsy, the decrease in uterine weight was not different in OVX and OVX + R rats. Ovariectomy also induced a significant decrease in both total and distal metaphyseal femoral mineral density, which was prevented by rutin consumption. Moreover, femoral failure load, which was not different in OVX and SH rats, was even higher in OVX + R rats than in OVX or SH rats. In the same way, on day 90, both urinary deoxypyridinoline (DPD) excretion (a marker for bone resorption) and calciuria were higher in OVX rats than in OVX + R or SH rats. Simultaneously, plasma osteocalcin (OC) concentration (a marker for osteoblastic activity) was higher in OVX + R rats than in SH rats. High-performance liquid chromatography (HPLC) profiles of plasma samples from OVX + R rats revealed that mean plasma concentration of active metabolites (quercetin and isorhamnetin) from rutin was 9.46+/-1 microM, whereas it was undetectable in SH and OVX rats. These results indicate that rutin (and/or its metabolites), which appeared devoid of any uterotrophic activity, inhibits ovariectomy-induced trabecular bone loss in rats, both by slowing down resorption and increasing osteoblastic activity.

Amylin and bone metabolism in streptozotocin-induced diabetic rats.

Amylin (AMY) is a 37 amino acid peptide cosecreted with insulin (INS) by pancreatic beta-cells and absent in type 1 diabetes, a condition frequently associated with osteopenia. AMY binds to calcitonin receptors, lowers plasma calcium concentration, inhibits osteoclast activity, and stimulates osteoblasts. In the present study, we examined the effects of AMY replacement on bone loss in a streptozotocin (STZ)-induced rodent model type 1 diabetes. Of 50 male Wistar rats studied, 40 were made diabetic with intraperitoneal STZ (50 mg/kg; plasma glucose concentrations > 11 mM within 5 days). Ten nondiabetic control (CONT) rats received citrate buffer without STZ. Diabetic rats were divided into four groups (n = 10/group) and injected subcutaneously with rat AMY (45 mg/kg), INS (12 U/kg), both (same doses), or saline (STZ; diabetic controls) once per day. After 40 days of treatment and five 24-h periods of urine collection for deoxypyridinoline (DPD), the animals were killed, blood was sampled, and femurs were removed. The left femur was tested for mechanical resistance (three-point bending). The right femur was tested for total, diaphyseal (cortical bone), and metaphyseal (trabecular bone) bone densities using dual-energy X-ray absorptiometry (DXA). Bone was ashed to determine total bone mineral (calcium) content. None of the treatments had any significant effect on femoral length and diameter. Untreated diabetic rats (STZ; 145+/-7N) had lower bone strength than did nondiabetic CONT (164+/-38; p < 0.05). Total bone mineral density (BMD; g/cm2) was significantly lower in STZ (0. 2523+/-0.0076) than in CONT (0.2826+/-0.0055), as were metaphyseal and diaphyseal densities. Diabetic rats treated with AMY, INS, or both had bone strengths and bone densities that were indistinguishable from those in nondiabetic CONT. Changes in bone mineral content paralleled those for total BMD (T-BMD). Plasma osteocalcin (OC) concentration, a marker for osteoblastic activity, was markedly lower in untreated diabetic rats (7. 6+/-0.9 ng/ml); p < 0.05) than in nondiabetic CONT (29.8+/-1.7; p < 0.05) or than in AMY (20.1+/-0.7; p < 0.05). Urinary DPD excretion, a marker for bone resorption, was similar in untreated and AMY-treated diabetic rats (35.0+/-3.1 vs. 35.1+/-4.4 nmol/mmol creatinine), intermediate in rats treated with INS (49.9+/-2.7), and normalized in diabetic rats treated with both agents (58.8+/-8.9 vs. 63.2+/-4.5 in CONT). Thus, in our STZ rat model of diabetic osteopenia, addition of AMY improved bone indices apparently by both inhibiting resorption and stimulating bone formation.

Parathyroid hormone (PTH) and PTH-related peptide induce relaxation of smooth muscle cells from guinea pig ileum: interaction with vasoactive intestinal peptide receptors.

PTH-related peptide (PTHrP), which shares 8 of 13 NH2-terminal residues with PTH, causes similar biological effects and interacts with the same receptor as PTH. In the gastrointestinal tract, human PTH and PTHrP-(1-34) relax rat fundic strips. However, the level of their action and the receptor involved in this effect are unknown. The aims of this study were 1) to determine the effects of human PTH-(1-34), human PTHrP-(1-34), -(1-16), and -(7-34) and vasoactive intestinal peptide (VIP) on circular isolated smooth muscle cells from guinea pig ileum; 2) to study the intracellular pathways involved in these effects; and 3) and to characterize the receptors involved by using specific antagonists. Smooth muscle cells were dispersed by enzymatic digestion. Contraction was assessed by measuring the length of 50 cells and expressed as the percent decrease in cell length from the control value. The relaxing effects of PTH, PTHrP and analogs, VIP, or antagonists were expressed as a percentage of the maximal effect observed in their absence. VIP, PTH-(1-34), and PTHrP-(1-34), -(1-16), and -(7-34) had no effect by themselves on these cells. However, when cells were contracted by the sulfated C-terminal octapeptide of cholecystokinin (10 nM), VIP, PTH-(1-34), and PTHrP(1-34) inhibited the sulfated C-terminal octapeptide of cholecystokinin-induced contraction in a concentration-dependent manner, whereas PTHrP-(1-16) and -(7-34) had no effect. The EC50 values of VIP, PTH-(1-34), and PTH-(1-34), and PTHrP-(1-34) were 7 nM, 20 pM, and 20 pM, respectively. The VIP antagonist ([D-P-Cl-Phe6,Leu17]VIP) inhibited VIP-, PTH-(1-34)-, and PTHrP(1-34)-induced relaxation, with IC50 values of 20, 500, and 400 pM, respectively. Likewise, the PTH/PTHrP antagonist [Tyr34-bovine PTH-(7-34)NH2] inhibited PTH-(1-34)-, PTHrP(1-34)-, and VIP-induced relaxation, with IC50 values of 1, 1, and 90 pM, respectively. Preincubation of cells with somatostatin, N-ethylmaleimide, and (R)-p-cyclic adenosine-3',5'-monophosphothioate inhibited the PTH-(1-34), PTHrP(1-34)-, and VIP-induced relaxation. In conclusion, human PTH and PTHrP induce a relaxation of intestinal smooth muscle by a direct myogenic effect. This effect requires the 1-34 amino acid sequence and is mediated by the activation of adenylate cyclase and protein kinase-A. Interactions among PTH, PTHrP, and VIP indicate that they may cross-react with their respective receptors.

Endurance training and bone metabolism in middle-aged rats.

This study was performed to observe the influence of moderate treadmill running on bone of middle-aged male rats. Seventy 15-month-old Wistar rats were used. Ten initial controls (IC) were killed on day 0. Among the 60 others, three groups of ten exercised rats (E) run 1 h/day, 6 days/week at 60% of their maximum aerobic capacity. On days 30, 60 and 90 of the training period, 20 rats, ten E and ten R (resting animals), were killed. Femoral failure stress never varied and was never different in E and R during the experiment. On day 90 whole body mineral content and mineral density were higher in E than R. Simultaneously, total, diaphyseal and metaphyseal femoral densities were lower in R than IC or than in E. No difference was observed between IC and E. In resting rats, urinary deoxypyridinoline excretion (a marker of bone resorption) increased between days 0 and 90, while it did not change in runners. These results indicate that in middle-aged rats, moderate running prevents decrease in bone mineral density, probably by inhibiting bone resorption.

Inhibition by calcitonin of hypercalcaemia induced by 1,25-dihydroxycholecalciferol.

The influence of injections of calcitonin on the hypercalcaemia and hyperhophosphataemia induced by 1 alpha-hydroxycholecalciferol (1alpha-OHD3) and 1,25-dihydroxycholecalciferol (1,25-(OH)2D3) was studied in growing lambs and lactating cows respectively. In lambs, the hypercalcaemic and hyperphosphataemic effect of 0.1 microgram 1 alpha-OHD3/kg body weight (i.v.) was completely inhibited by 20 units calcitonin/kg body weight, injected subcutaneously in four equal doses at intervals of 12 h, the first injection of calcitonin occurring immediately after injection of 1 alpha-OHD3. Moreover, the same dose of calcitonin partly inhibited the hypercalcaemia and hyperphosphataemia observed after injection of 0.25 microgram 1 alpha-OHD3/kg body weight. In lactating cows, four injections of calcitonin (5 u./kg body weight each, at intervals of 12 h) prevented the rise of plasma calcium and phosphate levels occurring after injection of 1,25-(OH)2D3 (0.01 microgram/kg body weight). The results indicated that high plasma calcitonin levels can inhibit the hypercalcaemic and hyperphosphataemic effects of 1,25-(OH)2D3 in ruminants. Thus, hypocalcaemia might occur in parturient cows in spite of raised concentrations of 1,25-(OH)2D3 in plasma.

Calcitonin may modulate placental transfer of calcium in ewes.

In pregnant ewes bearing twin fetuses and fed an adequate Ca diet, the consequences of calcitonin (CT) deficiency (induced by thyroidectomy performed on day 30 of gestation, associated with daily thyroxine supplementation) differed according to the time of pregnancy. Such a deficiency had no significant effect either on fetal total body Ca content or on placental transfer of Ca in 77-day-old fetuses. On the contrary, CT deficiency for 110 days (on day 140 of pregnancy) was associated with an increased Ca concentration in fetal total body and increased placental transfer of Ca from the dam to its fetus, which did not occur in five thyroidectomized ewes supplemented with thyroxine and CT. This indicates that CT might protect the skeleton of the pregnant female against excessive demineralization by modulating placental transfer of Ca, when such a transfer become important, during periods of intense mineralization of the fetal skeleton.

Prolactin and calcium metabolism in pregnant ewes.

The effect of ovine prolactin on intestinal Ca absorption and placental Ca transfer was studied in pregnant ewes. Six groups of five animals bearing a single fetus were used and injected s.c. daily between days 121 and 135. The first group was given 0.1 microgram ovine prolactin/kg body wt per day, the second 0.1 microgram ovine prolactin/kg body wt per day plus 0.1 microgram 1 alpha-hydroxycholecalciferol (1 alpha-OH-D3)/kg body wt per day, the third 0.1 microgram ovine prolactin/kg body wt per day plus 0.20 units calcitonin/kg body wt per day, the fourth 2 micrograms bromocriptine/kg body wt per day, the fifth 2 micrograms bromocriptine/kg body wt per day plus 0.1 microgram ovine prolactin/kg body wt per day, and the sixth were controls injected with vehicle alone. The intestinal Ca absorption was measured on a duodenal loop tied off in vivo on day 136 and placental Ca transfer was evaluated between days 129 and 136. Ovine prolactin stimulated both intestinal Ca absorption and placental Ca transfer; these effects were further increased by 1 alpha-OH-D3. Calcitonin had no effect on ovine prolactin-stimulated intestinal Ca absorption, but blunted the influence of ovine prolactin on Ca placental transfer. Bromocriptine decreased both intestinal Ca absorption and Ca placental transfer but these effects of bromocriptine were overcome by simultaneous injection of ovine prolactin.

Increase in plasma concentrations of atrial natriuretic peptide during infusion of hypertonic saline in conscious newborn calves.

Plasma concentrations of atrial natriuretic peptide (ANP), plasma renin activity (PRA), plasma concentrations of aldosterone, urine flow rate and sodium and potassium excretion were studied in two groups of four conscious 3-day-old male calves, infused with hypertonic saline or vehicle. Hypertonic saline infusion (20 mmol NaCl/kg body weight) was accompanied by a progressive rise in plasma concentrations of ANP (from 16.5 +/- 0.2 pmol/l at time 0 to 29.3 +/- 3.0 pmol/l at 30 min; P less than 0.05) and by a gradual decrease in PRA (from 1.61 +/- 0.23 nmol angiotensin I/l per h at time 0 to 0.54 +/- 15 nmol angiotensin I/l per h at 90 min; P less than 0.05); there was no change in the plasma concentration of aldosterone. Within the first 2 h of the 24-h urine collection period there was a marked rise in urine flow rate and sodium excretion in treated calves when compared with control animals (66.0 +/- 8.3 vs 15.9 +/- 1.2 ml/kg body weight per 2 h (P less than 0.05) and 6.7 +/- 1.3 vs 0.4 +/- 0.02 mmol/kg body weight per 2 h (P less than 0.01) respectively). During the following 22 h, urinary water and sodium excretion remained at significantly high levels. Thus, in the conscious newborn calf, changes in plasma ANP levels and urinary water and sodium excretion during hypertonic saline infusion are compatible with the hypothesis that endogenous ANP participates, at least in part, in the immediate diuretic and natriuretic renal response induced by the sodium overload.

1,25-Dihydroxycholecalciferol regulates salivary phosphate secretion in cattle.

The influence of 1,25-dihydroxycholecalciferol (1,25-(OH)2D3) on salivary inorganic phosphorus (Pi) concentration and secretion was studied in two groups of four heifers, the right parotid ducts of which were chronically fitted with a re-entrant cannula. In four heifers i.v. Pi loading (5 mmol/min for 2 h) induced hyperphosphataemia associated with a decrease in plasma 1,25-(OH)2D concentration and an increase in salivary Pi concentration and secretion. In four other heifers, daily 1 alpha-hydroxycholecalciferol injections (1 microgram/kg body wt per day for 3 days) induced hyperphosphataemia associated with an increase in plasma 1,25-(OH)2D concentration and a decrease in salivary Pi concentration and secretion. These treatments had no significant effect on salivary calcium concentration and secretion. Our results indicate that plasma 1,25-(OH)2D concentrations rather than phosphataemia regulate salivary Pi concentration and secretion in cattle.

Effect of a hypertonic saline load on plasma concentrations of atrial natriuretic peptide in fetal calves and their dams.

Plasma concentrations of atrial natriuretic peptide (ANP) were studied in eight adult non-pregnant cows and in two groups of six chronically catheterized bovine fetuses and their mothers in the eighth month of pregnancy. The first group of fetuses was used for studying the effect of an acute i.v. sodium load (240 mmol NaCl/fetus) on fetal ANP; the second group acted as controls. The mean basal ANP levels in the third-trimester bovine fetus were three to four times higher than maternal values (39.5 +/- 5.5 and 9.4 +/- 0.6 pmol/l respectively; P less than 0.01). Basal maternal plasma ANP levels were twice as high in pregnant cows in the third trimester of pregnancy than in non-pregnant cows (9.4 +/- 0.6 and 4.3 +/- 0.7 pmol/l respectively; P less than 0.05). In response to an i.v. hypertonic saline injection, fetal plasma ANP levels increased significantly (P less than 0.01) to a maximum of 86.7 +/- 17.6 pmol/l 10 min after the injection, and returned to baseline within 60 min after the treatment; during the 20 min following the i.v. sodium load, fetal plasma ANP correlated significantly with fetal plasma sodium concentrations (r = 0.96; n = 12) and with fetal plasma osmolality (r = 0.94; n = 12). No significant changes in maternal ANP values were observed in the two groups of animals. These results suggest that ANP secretion is stimulated during pregnancy in cows, and that, in the bovine fetus, a hypertonic sodium load appears to be a potent stimulus for ANP release.

Synthetic parathyroid hormone-related peptide (1-34) fragment stimulates placental calcium transfer in ewes.

The influence of synthetic parathyroid hormone related peptide (PTHrp) fragments on placental transfer of Ca was studied in four groups of four single ovine fetuses fitted with catheters chronically implanted into their left jugular vein (for injections) and carotid artery (for blood sampling), and used between days 104 and 118 of gestation. The first group received PTHrp(1-34), the second PTHrp(107-138), the third bovine PTH(1-34), and the last (control) group was injected with solvent alone. Each peptide (6 nmol/fetus per day) was injected i.v. three times per day from day 105 until day 116 of gestation. Placental Ca transfer (mmol/24 h per kg fetal wt) from the dam to the fetus was not different in control fetuses (7.1 +/- 0.6) and those given PTHrp(107-138) (7.2 +/- 0.5), but it was significantly increased by bovine PTH(1-34) (8.6 +/- 0.4; P less than 0.05) and by PTHrp(1-34) (10.1 +/- 0.3; P less than 0.01). Both peptides also significantly increased plasma concentrations of 1,25 dihydroxy-vitamin D3 (1,25-(OH)2D). These results indicate that PTHrp(1-34) can stimulate placental Ca transfer by increasing 1,25-(OH)2D synthesis, but also possibly by acting directly upon the placenta.

Aldosterone regulates salivary sodium secretion in cattle.

Intravenous infusion of aldosterone (10 microgram/kg body wt per h for 5 h) in four 2-month-old calves decreased salivary and urinary sodium (Na+) concentration and increased salivary potassium (K+) concentration without modifying salivary flow or urinary K+ concentration. Intravenous angiotensin II infusion (0.3 microgram/kg body wt per min for 1 h) in four Na+-replete 16-month-old bulls decreased salivary Na+ concentration and increased that of K+. It also increased plasma cortisol and plasma aldosterone concentrations, and decreased plasma renin activity (PRA). In four 16-month-old bulls Na+ deficiency (induced by chronic cannulation of the right parotid duct and loss of saliva for 5 days) had similar effects to those observed following aldosterone infusion in calves: a decrease in salivary Na+/K+ ratio. This decrease was associated with an increase in PRA and an increase in plasma aldosterone concentration. In these animals a close positive relationship was observed between PRA and plasma aldosterone concentration (r = 0.91; n = 20; P less than 0.01). Thus in cattle, during Na+ deficiency, the effect of aldosterone on parotid glands participates in the regulation of Na+ metabolism.

Parathyroid hormone-related peptide might stimulate calcium secretion into the milk of goats.

Parathyroid hormone-related peptide (PTHrP) has been shown to stimulate the placental calcium pump in pregnant ewes. Recent studies also suggest a physiological role for this peptide during lactation. In the present work, we have studied the possible role of synthetic human PTHrP(1-34), (1-86) and (140-173) fragments on calcium, inorganic phosphorus and magnesium secretion into milk of four adult goats in early lactation. This was done by measuring differences in calcium, inorganic phosphorus and magnesium concentrations in the carotid artery and superficial epigastric vein, the concentration of these minerals in milk and milk production. Both PTHrP(1-34) and (1-86) fragments stimulated calcium, inorganic phosphorus and magnesium uptake by the mammary gland and secretion into milk, without any significant effect on milk production. Therefore PTHrP might have important physiological roles in the pregnant and/or lactating female, by regulating calcium transport through the placenta and mammary gland.

Amylin inhibits ovariectomy-induced bone loss in rats.

Amylin (AMY), a peptide co-secreted with insulin by pancreatic beta-cells, inhibits bone resorption and stimulates osteoblastic activity. The ovariectomized (OVX) rat is an established animal model for human osteoporosis. Thus, the present experiment was performed to study the effects of AMY on estrogen deficiency-induced bone loss in rats. Thirty-one 6-month-old Wistar rats were randomized by body weight (BW) into two groups. The first underwent surgical OVX (n=21). The second was sham-operated (SH; n=10). Sixty days after surgery, 11 OVX rats were s.c. injected with rat AMY (3 microg/100 g BW/day, for 30 days; OVX+AMY), and 10 with solvent alone in the same way (0.15 ml/100 g BW; OVX). Each rat, housed in an individual cage, was fed daily the mean quantity of diet consumed the day before by SH rats. This diet contained 0.24% calcium and 0. 16% phosphorus. The 31 animals were killed on day 90. No difference in daily weight gain and BW was observed between groups. Neither AMY treatment nor OVX had any significant effect upon femoral morphology, femoral failure load, diaphyseal femoral density (representative of cortical bone) and total femoral calcium content. Nevertheless, both distal metaphyseal (representative of cancellous bone) and total femoral bone densities were higher in SH and OVX+AMY than in OVX rats. The highest plasma osteocalcin concentration was measured in OVX+AMY rats. Simultaneously, urinary deoxypyridinoline excretion was lower in OVX+AMY than in OVX rats. These results indicate that in OVX rats, AMY treatment inhibited trabecular bone loss both by inhibiting resorption and by stimulating osteoblastic activity.

Short communication: parathyroid hormone-related peptide does not stimulate phosphate placental transport.

It has been demonstrated that parathyroid hormone-related peptide (PTHrP) stimulates calcium (Ca) placental transport. Ca and inorganic phosphorous (P) metabolisms are intimately linked. Thus we have studied the influence of PTHrP on P placental transport in ewes. In our experimental paradigm, synthetic human PTHrP (1-86) fragment intravenously injected in six single chronically catheterized fetuses (3 x 1 micrograms/fetus/day, from day 129 to day 140 of gestation) had no significant effect upon P placental transport which was 131 +/- 6 and 129 +/- 9 mg/kg fetal weight/day in treated and control fetuses, respectively.

Acute versus chronic conditions for studies of fetal nutrient requirements: applicability to the human.

In order to validate results obtained in 'acute' versus 'chronic' experimental conditions, two fetal sheep nutrients, lactate and glucose, have been determined in 'acute' conditions and compared with the 'chronic' data in the literature. Maternal and fetal blood glucose in 'acute' conditions was in the range of published 'chronic' data. Fetal blood lactate was similar in 'acute' and during 'chronic' conditions. Maternal and fetal veno-arterial (VA) differences were in the same direction in 'acute' and in 'chronic' conditions for both lactate and glucose and of the same magnitude for lactate. For glucose, a good relationship was observed between umbilical VA differences and maternal arterial concentrations for all 'acute' and 'chronic' values. Thus no fundamental differences appear in the results obtained in 'acute' or during 'chronic' conditions. The applicability of such animal results to the human and the use of acute conditions to study fetal nutrition in the human are discussed.

Genistein and daidzein modulate in vitro rat uterine contractile activity.

The present study investigated the effect of genistein, daidzein and estradiol on in vitro rat uterine responsiveness to oxytocin (OT) and PGF(2)alpha or luprostiol (L). In a first experiment, animals were either sham-operated (SH; n=5), or ovariectomized (OVX; n=20) and orally treated for three months with either genistein (G; n=5; 10 microg/g BW/d) or daidzein (D; n=5; 10 microg/g BW/d) or 17 alpha-ethinylestradiol (E; n=5; 23 microg/kg BW/d) or untreated (OVX; n=5). At necropsy, the basal uterine tension was lower in OVX, G and D than in SH, the highest value being measured in E. Oxytocin (10(-12); 10(-11) M) or PGF(2)alpha (10(-12); 10(-9) M) induced an increase in SH, but not in OVX, E and G. In D, only the highest doses were efficient. In a second experiment, 20 intact animals were s.c. injected with either genistein (G; n=5; 10 microg/g BW) or daidzein (D; n=5; 10 microg/g BW) or estradiol benzoate (E; n=5; 23 microg/kg BW) or vehicle (C: controls; n=5), and killed 24 h later. In C and E, OT (10(-15) to 10(-10) M) or L (10(-12) to 10(-7) M) stimulated uterine contractile activity in a dose-dependent manner until a maximal level. On the opposite, in G and D, contractile agents (except the highest luprostiol doses) did not stimulate myometrium contractions. Moreover, radioligand binding assays showed that genistein or daidzein inhibited the specific binding of [(3)H] estradiol to the calf uterus estrogen receptor (ER). Therefore, it could be postulated that both genistein and daidzein might bind to the rat uterus ER, inducing either anti-estrogenic or very weak estrogenic effects (depending on the experimental conditions) on in vitro uterine responsiveness to OT and PGF(2)alpha or luprostiol.

Influence of clenbuterol on bone metabolism in exercised or sedentary rats.

This paper reports that the selective beta(2)-adrenergic receptor agonist clenbuterol affects bone metabolism in growing 3-mo-old male Wistar rats treated over 8 wk. Thirty-two 3-mo-old growing Wistar rats weighing 234 +/- 2 g were assigned to a progressive isometric force, strength-training exercise program plus oral clenbuterol (2 mg x kg body wt(-1) x day(-1)) for 5 days each week, exercise program without clenbuterol 5 days each week, no exercise program plus oral clenbuterol (2 mg x kg(-1) x day(-1)) for 5 days each week, or no exercise without clenbuterol 5 days each week. At the end of 8 wk, lean mass, fat mass, and right total femoral, distal metaphyseal femoral, and diaphyseal femoral bone mineral density were measured by Hologic QDR 4,500 dual X-ray absorptiometry (DEXA) technique. Left femoral bones were harvested after death on day 58, and femoral resistance was determined by three-point bending testing. We found that fat mass was decreased in rats given strength training exercise and decreased further in rats treated with clenbuterol. Lean mass was increased in clenbuterol-treated animals. Strength-training exercise appeared to have no effect on bone mineral density, serum osteocalcin, or urinary deoxypyridinoline. However, clenbuterol treatment decreased femoral length, diameter, bone mineral density, and mechanical resistance. Clenbuterol had no effect on osteocalcin but increased urinary deoxypyridinoline. We concluded that clenbuterol treatment decreased bone mineral density and increased bone resorption independent of the level of exercise rats were given.