Net Ca Absorption Research Articles

Calcium and vitamin D intake maintained from preovariectomy independently affect calcium metabolism and bone properties in Sprague Dawley rats.

The interaction of habitual Ca and vitamin D intake from preovariectomy to 4months postovariectomy on bone and Ca metabolism was assessed. Higher Ca intake suppressed net bone turnover, and both nutrients independently benefitted trabecular structure. Habitual intake of adequate Ca and ~50nmol/L vitamin D status is most beneficial. Dietary strategies to benefit bone are typically tested prior to or after menopause but not through menopause transition. We investigated the interaction of Ca and vitamin D status on Ca absorption, bone remodeling, Ca kinetics, and bone strength as rats transitioned through estrogen deficiency. Sprague Dawley rats were randomized at 8weeks to 0.2 or 1.0% Ca and 50, 100, or 1,000IU (1.25, 2.5, or 25μg) vitamin D/kg diet (2 × 3 factorial design) and ovariectomized at 12weeks. Urinary (45)Ca excretion from deep-labeled bone was used to assess net bone turnover weekly. Ca kinetics was performed between 25 and 28weeks. Rats were killed at 29weeks. Femoral and tibiae structure (by μCT), dynamic histomorphometry, and bone Ca content were assessed. Mean 25(OH)D for rats on the 50, 100, 1,000IU vitamin D/kg diet were 32, 54, and 175nmol/L, respectively. Higher Ca intake ameliorated net bone turnover, reduced fractional Ca absorption and bone resorption, and increased net Ca absorption. Tibial and femoral trabecular structures were enhanced independently by higher Ca and vitamin D intake. Tibial bone width and fracture resistance were enhanced by higher vitamin D intake. Dynamic histomorphometry in the tibia was not affected by either nutrient. A Ca × vitamin D interaction existed in femur length, tibial Ca content, and mass of the soft tissue/extracellular fluid compartment. Adequate Ca intake and serum 25(OH)D level of 50nmol/L provided the most benefit for bone health, mostly through independent effects of Ca and vitamin D.

The site of net absorption of Ca from the intestinal tract of growing pigs and effect of phytic acid, Ca level and Ca source on Ca digestibility

An experiment was conducted to test the hypothesis that the standardised digestibility of Ca in calcium carbonate and Lithothamnium calcareum Ca is not different regardless of the level of dietary Ca, and that phytic acid affects the digestibility of Ca in these two ingredients to the same degree. The objectives were to determine where in the intestinal tract Ca absorption takes place and if there are measurable quantities of basal endogenous Ca fluxes in the stomach, small intestine or large intestine. Diets contained calcium carbonate or L. calcareum Ca as the sole source of Ca, 0% or 1% phytic acid and 0.4% or 0.8% Ca. A Ca-free diet was also formulated and used to measure endogenous fluxes and losses of Ca. Nine growing pigs (initial body weight 23.8 ± 1.3 kg) were cannulated in the duodenum and in the distal ileum, and faecal, ileal and duodenal samples were collected. Duodenal endogenous fluxes of Ca were greater (p < 0.05) than ileal endogenous fluxes and total tract endogenous losses of Ca, but ileal endogenous fluxes were less (p < 0.05) than total tract endogenous losses. Standardised digestibility of Ca was not affected by the level of phytic acid, but decreased (p < 0.05) as Ca level increased in L. calcareum Ca diets, but that was not the case if calcium carbonate was the source of Ca (interaction, p < 0.05). The standardised duodenal digestibility (SDD), standardised ileal digestibility (SID) and standardised total tract digestibility (STTD) of Ca were not different if calcium carbonate was the source of dietary Ca. However, the STTD of Ca in L. calcareum Ca was greater (p < 0.05) than the SID and SDD of Ca. The SDD, SID and STTD of Ca in calcium carbonate were greater (p < 0.05) than those of L. calcareum Ca. In conclusion, under the conditions of this experiment, standardised digestibility of Ca is not affected by the level of phytic acid, but may be affected by dietary Ca level depending on the Ca source. Calcium from calcium carbonate is mostly absorbed before the duodenum, but Ca from L. calcareum Ca is mostly absorbed in the jejunum and ileum.

Effects of 25-hydroxyvitamin D3 on localisation and extent of gastrointestinal calcium absorption in dairy cattle

The combination of 25-hydroxyvitamin D3 (25-OHD3) and a diet negative in dietary cation anion difference (DCAD) has recently been shown to have beneficial effects on peripartal calcium (Ca) homeostasis in dairy cows. To further elucidate the underlying mechanisms, it was the aim of the experiments to investigate the effects of 25-OHD3 in combination with DCAD values about –70 meq/kg DM on pre-duodenal and overall gastrointestinal Ca absorption. A group of six ruminally fistulated lactating cows equipped with a cannula in the proximal duodenum were assigned to three dietary treatments (Control diet, anionic salts, anionic salts +25-OHD3) of 5 weeks each. Urine and faeces were collected quantitatively and flow of duodenal contents was calculated by applying chromium oxide. Blood samples were taken at regular intervals. Treatment with anionic salts and 25-OHD3 resulted in an increased Ca net absorption from the total gastrointestinal tract, which was mainly due to respective increases in intestinal Ca absorption. Furthermore, anionic salts and 25-OHD3-treated animals had significantly higher plasma phosphate concentrations and lower plasma levels of CrossLaps and the overall net absorption of phosphorus was significantly higher in these animals. From these data, it can be concluded that anionic salts in combination with 25-OHD3 positively influence the overall net Ca and phosphorus absorption, which is obviously associated with a reduced mobilisation of bone minerals as indicated by decreases in plasma CrossLaps concentrations.

Habitual calcium intake and vitamin D status during adulthood through estrogen deficiency have few interactions on calcium kinetics and bone

Eating patterns do not change as rapidly as the physiological changes that accompany the menopause. As such, habitual nutrition status may strongly affect post menopausal bone health in the general population. To test how dietary calcium (Ca) and vitamin D (VD) interact to influence bone during the menopause, we studied 70 female Sprague Dawley rats from young adulthood through ovariectomy (OVX)‐induced estrogen deficiency. Weanling rats were housed in a UV‐free environment and fed an AIN93G diet with 0.4% Ca and 100 IU VD/kg diet for 5 weeks. At 8 weeks of age, rats were randomized to AIN93M diets containing 0.2 or 1.0% Ca and 50, 100, or 1000 IU VD/kg diet. OVX was performed at 12 weeks of age. At 24–28 weeks of age, Ca kinetics was performed with a 45Ca tracer (oral Ca dose: 15 mg) up to 48 hours. Kinetics was modeled using WinSAAM software. A Ca and VD interaction was seen in the soft tissue compartment, but each nutrient independently increased endogenous Ca secretion. Bone formation was not affected by either nutrient. However, rats on the 1% Ca diet had lower fractional Ca absorption, higher net Ca absorption and higher bone balance due to reduced bone resorption, regardless of VD intake. Habitual Ca intake and VD status during adulthood throughout estrogen deficiency have few interactions on Ca kinetics and bone. Rather, Ca intake may be more influential to benefit bone than VD status at levels relevant to the general population.

Daily Supplementation with 25 µg Cholecalciferol Does Not Increase Calcium Absorption or Skeletal Retention in Adolescent Girls with Low Serum 25-Hydroxyvitamin D1–4

In healthy adolescents, cross-sectional studies show either no or negative relationships between serum 25-hydroxyvitamin D [25(OH)D] and calcium (Ca) absorption. Using a 2-period metabolic balance study, the effect of vitamin D supplementation on Ca absorption and retention in adolescent girls was investigated. Eleven girls aged 12–14 y with a mean entry serum 25(OH)D of 35.1 nmol/L consumed a controlled intake (providing 5 µg vitamin D and 1117 mg Ca/d) for two 3-wk metabolic balance periods separated by a 1-wk washout period. Sunlight exposure was minimized by sunscreen with a sun protection factor ≥ 15. After the first metabolic balance period, participants received 25 µg/d cholecalciferol supplementation for 4 wk. Fractional Ca absorption was measured in each metabolic balance period using a stable Ca isotope method. All urine and fecal samples were collected and analyzed to measure net Ca absorption and Ca retention. Paired t tests and correlations were used to analyze the data. Daily supplementation with 25 µg vitamin D resulted in a mean increase in serum 25(OH)D of 13.3 nmol/L (P < 0.01) but a decrease in fractional Ca absorption of 8.3% (P < 0.05) and no significant change in fasting serum 1,25-dihydroxyvitamin D, parathyroid hormone, net Ca absorption, or Ca skeletal retention. In pubertal girls with vitamin D status considered insufficient in adults, vitamin D supplementation of 25 µg/d for 4 wk did not improve fractional Ca absorption, net Ca absorption, or Ca retention.

Effects of isometric strength training followed by no exercise and Humulus lupulus L–enriched diet on bone metabolism in old female rats

We investigated in female rats the effects on bone metabolism of a prolonged no-training period, subsequent to an isometric exercise program, performed during young adulthood and those of a long-term consumption of Humulus lupulus L–enriched diet (genistein 1.92 and daidzein 1.24 mg/kg diet) combined or not with isometric training. Forty-eight rats (4 weeks old) were randomly divided into 4 groups: trained (C-Tr) or nontrained rats (C-NTr) fed with control diet and trained (H-Tr) or nontrained rats (H-NTr) fed with Humulus lupulus L–enriched diet. The diets lasted 100 weeks. Training was followed over a 25-week period. Bone parameters were measured at week 100. Our results showed that no significant difference was observed among the 4 groups in uterine relative weight, calcium (Ca) intake, fecal Ca, urinary Ca excretion, net Ca absorption, plasma Ca, and bone Ca content. Calcium balance was significantly enhanced in H-NTr rats in comparison with C-NTr and C-Tr rats. Isometric strength training led to a significant increase in total bone mineral density (BMD), diaphyseal BMD, and osteocalcin-deoxypyridinoline ratio in C-Tr rats compared with the other groups. The main findings of the present study indicate that in female rats, a 25-week isometric strength training performed during young adulthood followed by a prolonged no-training period increases BMD values and osteocalcin-deoxypyridinoline ratio, whereas long-term consumption of Humulus lupulus L–enriched diet does not improve bone parameters. It suggests that bone gains induced by exercise do not decrease immediately after cessation of training and also confirms the importance of the practice of physical activity during puberty and young adulthood to maximize the achieved peak bone density.

Fractional Calcium Absorption Is Increased in Girls with Rett Syndrome

Rett syndrome (RTT), an X-linked neurodevelopmental disorder primarilyaffecting girls, is characterized in part by osteopenia and increased risk of skeletal fractures. We hypothesized that decreased intestinal calcium (Ca) absorption relative to dietary Ca intake and increased renal Ca excretion might cause these problems in RTT. We measured fractional Ca absorption, urinary Ca loss, dietary Ca intake, and the hormonal factors regulating Ca metabolism to determine whether abnormalities in Ca balance might relate to poor bone mineralization in RTT girls and to evaluate the contribution of these factors to the overall dietary Ca needs of RTT girls. Ten RTT girls and 10 controls, matched for age, sex, and pubertal status, were given a 3 day constant Ca diet that mimicked their habitual intakes. At the end of each dietary period, girls received single doses of Ca (intravenous) and Ca (oral). Fractional urinary excretion of Ca, Ca, 24 hour urinary Ca, and urinary cortisol excretion were determined. Serum Ca, phosphorous, alkaline phosphatase, vitamin D metabolites, parathyroid hormone (PTH), and osteocalcin were measured in the postabsorptive state. Bone mineral content (BMC) was measured by dual-energy x-ray absorptiometry. Fractional Ca absorption was significantly higher in RTT than in control girls (mean +/- SDp, 52 vs. 33 +/- 13%). Dietary Ca intake (mean +/- SDp, 1,100 vs. 1,446 +/- 440 g/d) and net Ca absorption (mean +/- SDp, 513 vs. 362 +/- 306 mg/d) did not differ significantly between RTT and controls, respectively. Although urinary Ca excretion did not differ between groups, the increased urinary Ca:creatinine ratio (mean +/- SDp, 0.39 vs. 0.23 +/- 0.38) was consistent with clinical hypercalcuria and paralleled the significantly increased urinary cortisol excretion (mean +/- SDp, 3.1 vs. 1.7 +/- 1.1 mg/kg lean body mass per day) in the RTT girls. BMC was significantly lower in RTT than in controls (mean +/- SDp, 527 vs. 860 +/- 275 g). Serum Ca, P, alkaline phosphatase, vitamin D metabolites, PTH, and osteocalcin concentrations did not differ between the groups. Fractional Ca absorption showed a compensatory increase in the presence of adequate dietary Ca intakes, mild hypercalcuria, and pronounced bone mineral deficits in RTT girls. Whether supplemental dietary Ca could enhance fractional Ca absorption and improve bone mineralization in RTT girls is unknown.

Influence of myristic acid supplementation on energy, fatty acid and calcium metabolism of sheep as affected by dietary calcium and forage : concentrate ratio

In a 6 x 6 Latin square arrangement, sheep of 41 kg body weight were fed myristic acid [C14:0; 50 g/kg dry matter (DM)] supplemented to two basal diets of forage : concentrate ratios of 1 : 1.5 and 1 : 0.5 and adjusted to dietary calcium (Ca) contents of either 4.2 or 9.0 g/kg DM (the latter only together with C14:0 supplementation). Various variables of energy, fatty acid and Ca metabolism were determined in combined digestibility and respiratory chamber measurements. With C14:0 addition the energy loss via the faeces increased (p < 0.05, post hoc test) without affecting energy digestibility of the complete diet. The apparent digestibility of supplemented C14:0 was higher (p < 0.01) with approximately 0.8 in the forage-based diet than in the concentrate-based diet (approximately 0.6). The elevated levels of plasma C14:0 were mainly accompanied by reduced C18:0 and C18:1 levels. The estimated apparent content of metabolizable energy (ME) of added C14:0 was either 24.5 MJ/kg (concentrate-based diet) or 32.1 MJ/kg (forage-based diet). Extra Ca equalized these differences between basal diets and ME contents amounted to 33.0 MJ/kg on average. As expected from corresponding slight shifts in energy metabolizability, the total efficiency of ME utilization increased (p < 0.1) with C14:0. The lower level of dietary Ca was still within the range recommended, but adding C14:0 to the concentrate-based diet reduced Ca retention in the body of the sheep from 0.9 to -0.1 g/day because of an impaired (p < 0.05, post hoc test) net Ca absorption, whereas no effect was found with the forage-based diet. With C14:0 addition, plasma total phosphorus (P) and serum calcitrol levels increased (p < 0.05, post hoc test) while Ca concentrations did not clearly reflect the reduced net Ca absorption. Increasing the dietary Ca content prevented adverse effects on Ca retention in the concentrate-based diet and improved Ca retention in the forage-based diet. In conclusion, the C14:0 supplementation reduced Ca availability in concentrate-based diets while an additional supply of Ca improved Ca and energy retention. Consequently, Ca supply should exceed recommended levels in diet types where dietary lipids are likely to reduce Ca availability and a compromise in basal diet type has to found to be able to profit best from the energetic value and the methane-suppressing properties of C14:0.

Calcium kinetics are altered in clinically stable girls with cystic fibrosis.

Reduced bone mass in individuals with cystic fibrosis (CF) may result from alterations in calcium metabolism. Bone calcium deposition and resorption rates, calcium balance, and markers of bone turnover were assessed using stable isotopes of calcium in 22 prepubertal and pubertal girls with CF. Bone calcium deposition was associated with the availability of dietary calcium, total serum osteocalcin, and leptin concentrations. Reduced bone mass in individuals with CF may result from inadequate bone calcium (Ca) deposition, and excessive resorption, although these parameters have not been directly assessed in children with CF. We used stable Ca isotopes to measure rates of bone Ca deposition (Vo+), resorption, and retention in 22 clinically stable girls with CF (aged 7-18 yr). Rates of bone Ca deposition were determined by mathematically modeling the disappearance of iv Ca stable isotope ((42)Ca) for 6 d post dosing. Indirect markers of bone turnover and hormones associated with pubertal development were also assessed. Rates of bone Ca deposition and retention were highest during early puberty (Tanner stages 2 and 3). Calcium deposition rates in prepubertal (Tanner 1) and postmenarchal girls (Tanner stages 4 and 5) did not support substantial bone Ca retention. Net absorption of dietary Ca and serum osteocalcin and leptin concentrations were positively associated with Vo+. Time post menarche and serum leptin concentrations explained 91% of the variability in Vo+ (P = 0.0007). Serum total osteocalcin was low (10.9 +/- 5.4 ng/ml), and a substantial percentage of osteocalcin was undercarboxylated (54.3 +/- 11.8%). We concluded that increased calcium absorption and serum leptin concentrations were significantly associated with rates of bone Ca deposition, demonstrating an impact of nutritional status on this process. Rates of bone Ca deposition were lower than typically reported in healthy children, as were indirect markers of bone formation. These alterations in bone turnover contribute to reduced bone mass in girls with CF.

Melibiose, Difructose Anhydride III and Difructose Anhydride IV Enhance Net Calcium Absorption in Rat Small and Large Intestinal Epithelium by Increasing the Passage of Tight Junctions In Vitro

An Ussing chamber technique was used to determine the effects of three indigestible disaccharides on net Ca transport from the luminal side to the basolateral side of isolated preparations of jejunal, ileal, cecal and colonic epithelium in rats. Permeability of Lucifer Yellow (LY) and transepithelial electrical resistance (TEER), which are indicators of intercellular passage of the intestinal mucosa, were also determined. The concentrations of Ca in the serosal and mucosal media were 1.25 mmol/L and 10 mmol/L, respectively. After a 30-min incubation, the net Ca transport, LY passage and TEER were determined. In the control experiment, LY permeability was lowest, and TEER value was highest in the colon. The addition of 1-100 mmol/L melibiose, difructose anhydride (DFA)III, or DFAIV to the mucosal medium increased the net Ca absorption and LY permeability dose-dependently in the jejunum, ileum, cecum and colon preparations. Melibiose decreased TEER dose-dependently in the jejunum and cecum, but not in the ileum and colon. DFAIII decreased TEER dose-dependently in the jejunum, cecum and colon, but not in the ileum. DFAIV decreased TEER dose-dependently in all four intestinal portions. Positive linear relationships were found between net Ca transport and LY passage in all portions of the intestine, whereas negative linear relationships were found between net Ca absorption and TEER. We concluded that the three indigestible saccharides directly affect the epithelial tissue and activate the passage of tight junctions, thereby promoting Ca absorption in the small and large intestine in vitro.

Regional bone loss after orthotopic liver transplantation in inbred rats: the role of hepatic denervation.

Bone loss and long-term persistence of osteoporosis with increased fracture risk are common after liver transplantation. It is unknown whether transplantation-induced disruption of hepatic nerves, serving numerous regulatory metabolic and sensory functions, is herein involved. To test this possibility, we measured bone mineral density (BMD) by peripheral quantitative computed tomography (pQCT) and studied dynamic histomorphometry, radiocalcium kinetics, and biochemical parameters in 7 liver-transplanted and 7 sham-operated inbred rats. Although liver function was normal in TX rats, trabecular BMD of the first lumbar vertebra and total BMD of the femoral diaphysis were decreased by 13% and 6%, respectively, 9 months postsurgery. The breaking force of the femur was significantly lower by 21%. However, bone mass in the femoral and tibial metaphysis was preserved as evidenced by pQCT measurements and histomorphometry. Trabecular width and wall thickness were significantly decreased in vertebral cancellous bone, whereas indices of bone formation and resorption were normal or slightly reduced. Serum minerals, mineral balance, fractional and net absorption of Ca and Mg, serum calciotropic hormones, IGF-I, leptin, specific activity of 45Ca in bone, 45Ca excretion, and biochemical indices of bone formation and bone resorption remained unchanged. We conclude that liver transplantation-related denervation causes cancellous and cortical bone loss in well-innervated bone sites such as the lumbar spine and the long bone diaphysis. Cancellous bone loss in TX rats is due to an impairment of osteoblast team performance and subsequent trabecular thinning. The mechanism uncovered by our study may contribute to long-term bone loss after liver transplantation.

Various Indigestible Saccharides Enhance Net Calcium Transport from the Epithelium of the Small and Large Intestine of Rats In Vitro

An Ussing chamber technique was used to determine the effects of six indigestible saccharides on net Ca absorption from the luminal side to the basolateral side of isolated preparations of rat jejunal, ileal, cecal and colonic epithelium in vitro. The concentrations of Ca in the Tris buffer solution on the serosal side and on the mucosal side were 1.25 and 10 mmol/L, respectively. After a 30-min incubation, the Ca concentration in the serosal medium was determined and the net transepithelial Ca transport was calculated. The addition of 0.1-200 mmol/L maltitol, difructose anhydride (DFA)III, DFAIV, raffinose, fructooligosaccharide (FOS) or polydextrose (PD) to the mucosal medium increased the net Ca absorption dose-dependently in the jejunum, ileum, cecum and colon preparations. The threshold concentration required to enhance Ca transport and the extent of enhancement of Ca transport differed among the saccharides tested and among the portions of the intestine examined. Among the saccharides tested, DFA IV had the strongest effect on Ca absorption in the jejunum and cecum. We conclude that indigestible carbohydrates directly affect the epithelial tissue and promote Ca absorption in both the small and large intestine in vitro.

Short-chain fatty acids enhance diffusional Ca transport in the epithelium of the rat cecum and colon

We examined the effect of short-chain fatty acids (SCFAs) on Ca absorption from the large intestine in rats in vitro. An Ussing-type chamber technique was used to determine the net transport of Ca from the luminal side to the basolateral side of isolated epithelium in cecum and colon preparations. The concentration of Ca in the serosal and mucosal Tris buffer solution was 1.25 mM and 10 mM, respectively. Both solutions were warmed at 37 °C and bubbled with 95%O 2 and 5%CO 2. During and after the incubation period (30 min or 60 min), the Ca concentration in the serosal medium was determined and the net transepithelial Ca transport was evaluated. The addition of 80 mM acetic acid, 40 mM propionic acid and 10 mM butyric acid to the mucosal medium increased net Ca absorption (about 300%) in the cecum and colon. An individual application of acetic, propionic or butyric acid (0.01 to 100 mM) to the mucosal medium also increased net Ca absorption at doses of 10 mM and /or 100 mM in the cecum and colon. An increase in solute concentration in the mucosal medium by addition of glycerol or PGE400, or a decrease in pH (7.0–3.0) by addition of HCl did not affect transepithelial Ca transport. We concluded that SCFAs affect the epithelial tissue and promote Ca absorption from the large intestine in vitro. The enhancement of Ca transport induced by SCFAs might be involved in the paracellular transport mechanism.

The effect of high salt and high protein intake on calcium metabolism, bone composition and bone resorption in the rat

The effects of salt (NaCl) supplementation of rat diets (50 g/kg diet), with normal (200 g/kg) or high (500 g/kg) dietary casein content, were studied in 3-week-old male rats over a 3-week period. Weight gain was reduced by dietary salt but was unaffected by dietary casein. Salt-supplemented rats exhibited a two- and three-fold increase in urinary Mg and Ca excretion respectively, irrespective of dietary casein content. Dietary casein had no effect on urinary Ca or Mg. Salt reduced femoral mass but not femoral mass expressed relative to body weight, but neither variable was affected by dietary casein. Femoral Mg and P contents and concentrations were unaffected by dietary salt or casein. While femoral Ca concentration was unaffected by dietary salt, the Ca content was reduced by salt supplementation, irrespective of dietary casein content. Neither the content nor concentration of Ca in femora was affected by dietary casein. Urinary pyridinoline and deoxypyridinoline levels were increased by salt supplementation, irrespective of dietary casein content, but were unaffected by casein. Net Ca absorption was unaffected by dietary salt or casein. In conclusion, these results show that salt supplementation over the short-term increased the rate of bone resorption in rats. This was as a consequence of Na-induced calciuria. On the other hand, a high dietary protein intake had no effect on Ca metabolism, bone composition or bone resorption, nor did it augment the Na-induced calciuria or increased rate of bone resorption.

Acute effects of prolactin on passive calcium absorption in the small intestine by in vivo perfusion technique

The acute effect of intraperitoneally administered prolactin (0.2, 0.4, and 0.6 mg/kg body weight) on passive calcium transport in duodenum, proximal jejunum, and ileum of sexually mature female Wistar rats was investigated by using an in vivo perfusion technique. Test solution containing (in mM) NaCl, 100; KCl, 4.7; MgSO4, 1.2; CaCl2, 20; D-glucose, 11; sodium ferrocyanide (Na4Fe(CN)6), an index of net water transport, 20; and 0.7 microCi 45CaCl2 (1 Ci = 37 GBq) was perfused through the 10-cm intestinal loop for 60 min. Results showed that 0.4 mg prolactin/kg body weight significantly increased duodenal net Ca absorption (net Ca) from 23.81 +/- 1.84 to 30.56 +/- 1.57 mmol/g dry weight (p < 0.05) by stimulating the lumen to plasma calcium flux (CaL-P). The jejunum responded to 0.2, 0.4, and 0.6 mg prolactin/kg body weight by reversing from net Ca absorption of 18.60 +/- 1.70 mmol/g dry weight to net secretion of -3.30 +/- 1.56, -10.39 +/- 2.21, and -11.79 +/- 2.04 mmol/g dry weight (p < 0.01), respectively, as a result of a dose-dependent increase in plasma to lumen calcium flux (CaP-L). Calcium fluxes in the ileum on the other hand did not respond to prolactin. There was a close correlation between net water flux and net calcium flux in all three intestinal segments under basal condition regardless of the luminal sodium concentration. However, this correlation was lost after prolactin administration, which while having no effect on net water flux, altered the duodenal and jejunal calcium fluxes. By varying the luminal concentration of sodium, it was found that the stimulatory effect of 0.4 mg prolactin/kg body weight on the duodenal CaL-P was reduced when compared with control, i.e., 17.84 +/- 0.91 vs. 26.64 +/- 1.05 mmol/g dry weight at a sodium concentration of 180 mM, and 14.48 +/- 0.99 vs. 20.12 +/- 1.34 mmol/g dry weight at a sodium concentration of 140 mM. At a sodium concentration of 80 mM, the prolactin effect was absent. Since duodenal Na(+)-K+ ATPase activity was increased by prolactin from 3.77 +/- 0.16 to 4.95 +/- 0.30 mumol Pi.mg-1 protein.h-1 (p < 0.05), sodium dependency of the prolactin-enhanced lumen to plasma calcium flux may be related to both sodium-induced water flow and calcium-sodium exchange across the basolateral membrane. Thus, it was postulated that under basal condition, net calcium transport in the small intestine occurred with the sodium-induced water transport along the paracellular pathway. However, after prolactin administration, this association was lost. Prolactin-enhanced lumen to plasma calcium flux in the duodenum was sodium dependent and involved the Na(+)-K+ ATPase activity. In the proximal jejunum, prolactin stimulated plasma to lumen calcium flux, but the mechanism was not known.

Acute effects of prolactin on passive calcium absorption in the small intestine by in vivo perfusion technique

The acute effect of intraperitoneally administered prolactin (0.2, 0.4, and 0.6 mg/kg body weight) on passive calcium transport in duodenum, proximal jejunum, and ileum of sexually mature female Wistar rats was investigated by using an in vivo perfusion technique. Test solution containing (in mM) NaCl, 100; KCl, 4.7; MgSO4, 1.2; CaCl2, 20; D-glucose, 11; sodium ferrocyanide (Na4Fe(CN)6), an index of net water transport, 20; and 0.7 microCi 45CaCl2 (1 Ci = 37 GBq) was perfused through the 10-cm intestinal loop for 60 min. Results showed that 0.4 mg prolactin/kg body weight significantly increased duodenal net Ca absorption (net Ca) from 23.81 +/- 1.84 to 30.56 +/- 1.57 mmol/g dry weight (p < 0.05) by stimulating the lumen to plasma calcium flux (CaL-P). The jejunum responded to 0.2, 0.4, and 0.6 mg prolactin/kg body weight by reversing from net Ca absorption of 18.60 +/- 1.70 mmol/g dry weight to net secretion of -3.30 +/- 1.56, -10.39 +/- 2.21, and -11.79 +/- 2.04 mmol/g dry weight (p < 0.01), respectively, as a result of a dose-dependent increase in plasma to lumen calcium flux (CaP-L). Calcium fluxes in the ileum on the other hand did not respond to prolactin. There was a close correlation between net water flux and net calcium flux in all three intestinal segments under basal condition regardless of the luminal sodium concentration. However, this correlation was lost after prolactin administration, which while having no effect on net water flux, altered the duodenal and jejunal calcium fluxes. By varying the luminal concentration of sodium, it was found that the stimulatory effect of 0.4 mg prolactin/kg body weight on the duodenal CaL-P was reduced when compared with control, i.e., 17.84 +/- 0.91 vs. 26.64 +/- 1.05 mmol/g dry weight at a sodium concentration of 180 mM, and 14.48 +/- 0.99 vs. 20.12 +/- 1.34 mmol/g dry weight at a sodium concentration of 140 mM. At a sodium concentration of 80 mM, the prolactin effect was absent. Since duodenal Na(+)-K+ ATPase activity was increased by prolactin from 3.77 +/- 0.16 to 4.95 +/- 0.30 mumol Pi.mg-1 protein.h-1 (p < 0.05), sodium dependency of the prolactin-enhanced lumen to plasma calcium flux may be related to both sodium-induced water flow and calcium-sodium exchange across the basolateral membrane. Thus, it was postulated that under basal condition, net calcium transport in the small intestine occurred with the sodium-induced water transport along the paracellular pathway. However, after prolactin administration, this association was lost. Prolactin-enhanced lumen to plasma calcium flux in the duodenum was sodium dependent and involved the Na(+)-K+ ATPase activity. In the proximal jejunum, prolactin stimulated plasma to lumen calcium flux, but the mechanism was not known.

Calcium absorption, endogenous excretion, and endocrine changes during and after long-term bed rest

Negative calcium balance is a known consequence of bed rest, and is manifested in elevated urine and fecal calcium (Ca). Elevated fecal Ca can result from either decreased absorption, increased endogenous fecal excretion, or both. We measured the Ca absorption and endogenous fecal excretion in eight healthy male volunteers before and during 4 months of bed rest. Dual isotope (n = 6) or single isotope (n = 2) methods in conjunction with Ca balance were used to calculate true and net Ca absorption and endogenous fecal excretion. Stool Ca increased from 797 mg/day (mean intake 991 mg/day) to 911 mg/day during bed rest, whereas urine Ca excretion increased from 174 to 241 mg/day. True Ca absorption decreased from 31 +/- 7% of Ca intake pre-bed rest to 24 +/- 2% during bed rest, (p < 0.05) and returned toward pre-bed rest values within 5-6 weeks following reambulation. Endogenous fecal excretion did not change significantly, and therefore, most of the increased fecal Ca resulted from changes in absorption. However, in one individual, endogenous fecal Ca excretion was the major contributor to Ca loss. Ionized Ca and pyridinium crosslinks increased and 1,25(OH)2 vitamin D decreased during bed rest, similar to the decrease in Ca absorption; parathyroid hormone (PTH), calcitonin, serum albumin, phosphorus, and total serum Ca were unchanged. Although alkaline phosphatase, osteocalcin, and PTH were unchanged during bed rest, they were elevated during reambulation. These changes accompanied by increased Ca absorption and balance and decreased ionized and total serum Ca suggest a rebound in bone formation following immobilization.

Calcium Absorption, Endogenous Excretion, and Endocrine Changes During and After Long-Term Bed Rest

Negative calcium balance is a known consequence of bed rest, and is manifested in elevated urine and fecal calcium (Ca). Elevated fecal Ca can result from either decreased absorption, increased endogenous fecal excretion, or both. We measured the Ca absorption and endogenous fecal excretion in eight healthy male volunteers before and during 4 months of bed rest. Dual isotope (n = 6) or single isotope (n = 2) methods in conjunction with Ca balance were used to calculate true and net Ca absorption and endogenous fecal excretion. Stool Ca increased from 797 mg/day (mean intake 991 mg/day) to 911 mg/day during bed rest, whereas urine Ca excretion increased from 174 to 241 mg/day. True Ca absorption decreased from 31 +/- 7% of Ca intake pre-bed rest to 24 +/- 2% during bed rest, (p < 0.05) and returned toward pre-bed rest values within 5-6 weeks following reambulation. Endogenous fecal excretion did not change significantly, and therefore, most of the increased fecal Ca resulted from changes in absorption. However, in one individual, endogenous fecal Ca excretion was the major contributor to Ca loss. Ionized Ca and pyridinium crosslinks increased and 1,25(OH)2 vitamin D decreased during bed rest, similar to the decrease in Ca absorption; parathyroid hormone (PTH), calcitonin, serum albumin, phosphorus, and total serum Ca were unchanged. Although alkaline phosphatase, osteocalcin, and PTH were unchanged during bed rest, they were elevated during reambulation. These changes accompanied by increased Ca absorption and balance and decreased ionized and total serum Ca suggest a rebound in bone formation following immobilization.

Calcium and protein kinase C play an important role in Campylobacter jejuni-induced changes in Na + and Cl − transport in rat ileum in vitro

The pathophysiological mechanism of Campylobacter jejuni (enterotoxigenic) induced secretory diarrhoea remains least understood. To investigate the mechanism(s) involved, the unidirectional fluxes of Na + and Cl − were measured across the C. jejuni live culture infected and control (non infected) rat ileum (unstriped), in vitro by Ussing technique under short circuit conditions, in the presence or absence of: Ca 2+ ionophore A23187 (5 μM), 1-verapamil (100 μM), calmodulin (CaM) antagonist W-7 (100 μM), dantrolene (25 μM), protein kinase C (PKC) activator PMA (100 ng/ml) and H-7 (60 μM), selective inhibitor of PKC. There was net absorption of Na + and enhanced Cl − secretion in infected animals while in control animals there was net absorption of Na + and marginal secretion Cl −. Ca 2+ ionophore A23187 mimicked the effects of C. jejuni infection whereas 1-verapamil had significant antisecretory effect on Na + and Cl − secretion in infected animals. In vitro measurement of unidirectional 45Ca fluxes in Ussing chamber experiments revealed net absorption of Ca 2+ in infected rat ileum as compared to net secretion of Ca 2+ in control rat ileum. These observations clearly indicate that there is increased stimulation of Ca 2+ uptake from extracellular milieu to the enterocytes during C. jejuni-induced diarrhoea. The intracellular calcium levels (Ca 2+] i (as measured by fluorescent probe Fura-2AM) were found to be raised significantly ( P < 0.001) in enterocytes isolated from C. jejuni infected ileum as compared to the enterocytes from control ileum. The observed increase in [Ca 2+] i in enterocytes isolated from C. jejuni live culture supernatant treated rat ileum further shows the involvement of enterotoxin in diarrhoeal process. Dantrolene decreased significantly C. jejuni-induced net Na + and Cl − secretion but it could not reverse it to absorption suggesting the partial involvement of Ca 2+ mobilised from intracellular stores in mediating secretion. W-7 failed to inhibit the C. jejuni-induced net Na + and Cl − secretion. In addition the CaM activity estimated in intestinal microvillar core remained same in both the control and C. jejuni infected animals. This indicates that C. jejuni-induced diarrhoea is not mediated through the activation of Ca 2+-CaM complex pathway of the Ca 2+ messenger system. The PKC activator PMA, induced net secretion of Na + and Cl − in the control animals but it could not enhance further the C. jejuni-induced Na + and Cl − secretion, suggesting that there is overlapping effect of PMA and C. jejuni live culture infection. H-7 significantly inhibited ( P < 0.001) net Na + and Cl − secretion in the infected animals but had no effect on basal rat ileal Na + and Cl − transport in control animals. This again suggest the activation of PKC signal in C. jejuni-induced ileal Na + and Cl − secretion. Although intracellular cAMP levels were found to be significantly raised in the enterocytes isolated from C. jejuni live culture infected and C. jejuni culture supernatant treated rat ileum, the mechanism of interaction between cAMP and Ca 2+ messenger system in infection remains undefined. Taken together, the present results therefore, indicate that enterotoxigenic C. jejuni live culture-induced Na + and Cl − secretion is a calcium-dependent process where both increased Ca 2+ entry from extracellular milieu and Ca 2+ mobilization from intracellular stores result in increased enterocyte (Ca 2+] i. The activation of PKC further appears to be the important intracellular mediator pathway involved in Ca 2+-dependent stimulation of C. jejuni-induced diarrhoea.

Postnatal attainment of intrauterine macromineral accretion rates in low birth weight infants fed fortified human milk

Hypothesis: Provision of more bioavailable mineral sources as human milk supplements enables very low birth weight (VLBW) infants to meet the intrauterine accretion rate for calcium and phosphorus. Design: Comparison of currently formulated human milk fortifier with previous formulation. Setting: Neonatal level II and III nurseries Patients: Twenty-six healthy, VLBW infants, whose mothers chose to breast-feed. Interventions: We tested the effects of two formulations designed for VLBW infants as human milk supplements and differing primarily in their quantity and source of Ca, P, and magnesium. The study interval began with a milk intake of 100 ml · kg -1 · day -1 and ended when a body weight reached 2.0 kg. Main outcome measures: Net absorption and retention of Ca, P, and Mg during a nutritional balance study conducted once during the study interval, growth during the entire study interval, and bone mineral content of the radius were measured at the beginning and end of the study interval. Results: The newer Ca gluconate-glycerophosphate preparation (given to group CaGP) resulted in greater net absorption and retention of Ca and P ( p <0.01) than in infants given Ca phosphate (group CaTB). Mg retention was greater than (in group CaGP) or equivalent to (in group CaTB) the intrauterine accretion rate. Radius bone mineral content was significantly greater in group CaGP than in group CaTB ( p <0.001). Volumes of the fortified human milk preparation needed to meet the needs for gain in body weight were higher in group CaGP than in group CaTB ( p <0.001). Conclusions: Intrauterine accretion rates for Ca and P can be achieved when VLBW infants are fed human milk supplemented with Ca gluconate-glycerophosphate. Supplementation of human milk with Mg may not be indicated. In this study, greater intakes of Ca and P, and not improvements in bioavailability, result in improved net retention and bone mineral content of VLBW infants. (J P ediatr 1995;126:441-7)