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Biomedical subjects

A Rubinacci

Publications and source records attributed to A Rubinacci.

At least 37 records · Page 2Linked to original sources

Effects of amylin on human osteoblast-like cells.

Amylin has been reported to have bone-conserving effects. In the present study we evaluated the possible activity of the peptide on human osteoblast-like (hOB) cells in primary culture. Amylin between 10(-9) and 10(-6) M, dose-dependently stimulated cell proliferation with a maximal effect (200%) at 10(-6) M. In addition, amylin increased osteocalcin production when hOB cells were exposed to 1,25(OH)2D3 (10(-8)M) but there was a nonsignificant upward trend on alkaline phosphatase activity. The present results suggest that amylin could be included among the group of peptides endowed with osteogenic activity.

Amyloid↗

Effect of short course of 1,25-dihydroxyvitamin D3 on biochemical markers of bone remodelling in postmenopausal women.

This study was designed to test the hypothesis that a short treatment course of 1,25(OH)2D3 elicits a stimulation of osteoblast activity without any action on the osteoclast. To test this, oral daily doses of 0.5 microgram or 1 microgram of 1,25(OH)2D3 were administered for 7 days to two groups (n = 5 and n = 7, respectively) of postmenopausal women with low bone mineral density. Markers of osteoblast activity, i.e. osteocalcin (BGP), total alkaline phosphatase activity (ALP) and bone alkaline phosphatase activity (BALP), and markers of osteoclast activity, i.e. hydroxylysyl-pyridinoline (Pyr), lysyl-pyridinoline (D-Pyr), and galactosyl-hydroxylysine (GHyl) were measured in plasma and in fasting urinary samples, respectively, at sequential times during and after 1,25(OH)2D3 administration. It resulted that short term 1 microgram 1,25(OH)2D3 oral administration induced a significant (P < 0.05) rise of BGP serum level without any associated increase of D-Pyr and GHyl, the latter also expressed as GHyl to GGHyl ratio. Urinary Pyr increased significantly after 1 microgram daily doses of 1,25(OH)2D3. Thus, a short course of 1 microgram daily doses of 1,25(OH)2D3 elicits a stimulation of osteoblast activity without any enhancement of D-Pyr, the most specific marker of osteoclast activity. The enhancement of Pyr after 1 microgram daily doses of 1,25(OH)2D3 might be due to the activation of extraosseous metabolic pathways rather than to the activation of osteoclast.

Aged↗

Bicarbonate dependence of ion current in damaged bone.

The aim of this work was to characterize the ion current that enters mouse metatarsal bones following damage to the cortex. We assessed both the spatial distribution of this current and its dependence on the presence of bicarbonate in the medium. We used a voltage-sensitive probe system vibrating in two dimensions and recorded the signal as function of the position of the probe with respect to the site of damage and of ion substitutions in the medium. When the cortex was damaged (50 microm cylindrical hole penetrating into the marrow cavity), we recorded a steady state net inward electrical current directed toward the site of damage. In nonbicarbonate media, the density of the current was maximal near the center of the hole and ranged from 6 to 18 microA/cm2. As the probe was moved off the center of the hole, measured current density decreased in a manner consistent with the hypothesis that the source of the inward current is localized to the hole. After changing bicarbonate concentration in the medium from 0 to 42 mM, the current density nearly doubled, then decayed back to its original level exponentially over 35 minutes. When the diaphysis of living bone was left intact the current density was close to background level either in the presence or absence of bicarbonate in the medium. Damaged dead bone did not drive any current higher than background level. We conclude that the vibrating probe technique is a powerful tool to characterize ion currents in injured bone, helping to understand the physiology of bone-plasma interface and the bone healing processes. The current density transiently doubled upon addition of bicarbonate, indicating that this ion may carry the electrical current in damaged bone, probably by pump-leak mechanisms operating at the bone-plasma interface.

Animals↗

Effect of an oral calcium load on urinary markers of collagen breakdown.

Aim of this study was to investigate whether osteoclast activity changes as a consequence of even mild physiological perturbation of plasma calcium as such induced by an oral calcium load. Osteoclast activity was determined indirectly by measuring, in spot urines at two and four hours after oral calcium load, the urinary excretion of hydroxylysylpyridinoline (Pyr), deoxylysylpyridinoline (D-Pyr), hydroxyproline (Hyp) and galactosyl-hydroxylysine (GHyl). The occurrence of the metabolic perturbation of plasma calcium homeostasis was assessed by measuring three indexes: i.e. calcemic response, PTH reduction and calciuric response at times following oral calcium loading. A significant fall of urinary D-Pyr and Pyr followed the perturbation of calcium homeostasis induced by the oral calcium load in two groups of healthy young adult and postmenopausal women. The highest mean percent reduction was observed for D-Pyr and was quantitatively similar in the two groups. Since urinary D-Pyr is the most specific bone resorption marker, it may be inferred that the perturbation of plasma calcium homeostasis induced by an oral calcium load is able to acutely inhibit osteoclast activity. This supports the view that osteoclasts are involved in the short-term error correction of plasma calcium.

Adult↗

Galactosylhydroxylysine and pyridinium cross links in monitoring the bone response to hormone replacement therapy.

Aim of the study was to compare urinary galactosyl-hydroxylysine (GHyl), deoxypyridinoline (D-Pyr) and pyridinoline (Pyr) before and after 5 to 9 months of hormone replacement therapy (HRT) in postmenopausal women. The urinary markers were measured by HPLC in the second void of fasting samples and were expressed as ratio to creatinine. GHyl was also expressed as a ratio to glucosylgalactolysyl-hydroxylysine (GGHyl). After short-term hormone replacement therapy, urinary D-Pyr fell significantly, but Pyr and GHyl, also when expressed as a ratio to GGHyl, remained unmodified. We conclude that GHyl and Pyr are not useful markers in monitoring the bone response to HRT in postmenopausal women.

Alkaline Phosphatase↗

Comparison between urinary pyridinium cross-links and hydroxylysine glycosides in monitoring the effects of ovariectomy and 17 beta-estradiol replacement in aged rats.

This study was undertaken to assess the sensitivity of hydroxylysylpyridinoline (HP), lysylpyridinoline (LP), galactosylhydroxylysine (GHyl) and glucosylgalactosylhydroxylysine (GGHyl) to monitor bone response to estrogen deficiency and replacement by comparing their excretory patterns in ovariectomized aged (11-14 months old) rats. The ovariectomized (OVX) rats were randomized into two groups: (1) OVX plus vehicle; (2) OVX plus 17 beta-estradiol (17-beta E, 10 micrograms/kg, s.c., 4 days/week). Treatment with 17-beta E started immediately after OVX and continued for 60 days. The collagen catabolites were measured in urine for 1 month before OVX and thereafter for 60 days. In temporal coincidence with urine collection, bone area and bone mineral density (BMD) of lumbar vertebrae, femoral diaphysis and distal metaphysis were measured by dual-energy X-ray absorptiometry. In the untreated rats, BMD of the femoral metaphysis and lumbar vertebrae decreased significantly and the urinary excretion of LP, HP, GHyl and GGHyl increased with different patterns. In the treated rats, 17-beta E replacement prevented the increment in LP excretion, partially prevented the increase in HP excretion, but had no effect on the excretion of GHyl and GGHyl. In conclusion pyridinolines and glycosides have different sensitivities to the bone response to OVX. Glycoside excretion after OVX also reflects metabolic processes not strictly related to bone loss and, in contrast with LP, is not sensitive to estrogen replacement.

Absorptiometry, Photon↗

Osteocalcin production in vivo and in vitro after 1,25-dihydroxycholecalciferol stimulation comparison of different assays.

The study was designed to assess the sensitivity of three commercial assays (which differ in methodology, standard and antibodies) for osteocalcin, used for detecting changes in osteocalcin secretion induced by calcitriol (1,25-dihydroxycholecalciferol) in vivo and in vitro. Osteocalcin levels were determined in serum samples of 10 osteoporotic women after short term calcitriol treatment, and in the culture medium of human osteoblast-like cells (n = 22) after 48 h calcitriol exposure. All assays displayed similar sensitivity in detecting osteocalcin production in vivo after a 1 microgram daily dose of calcitriol. A novel IRMA (CIS), claimed to detect intact osteocalcin, showed higher osteocalcin values than the other assays, and in vitro showed the best sensitivity; it provides an appropriate index of the osteocalcin synthetic activity of cultured human osteoblasts.

Aged↗

Time-relationship between bone growth and increment of bone mineral content in growing rats.

The aim of the study was to describe the time-relationship between the growth rate of the rat tibia and the increase rate of its mineral content. Appositional and endochondral bone growth rates were derived from sequential X-rays measurements of tibial widening and lengthening, respectively; the increase rate of bone mineral content was derived from sequential photon absorptiometry measurements of the proximal tibio-fibular site. The time-relationship of appositional growth rate and endochondral growth rate versus bone mineral content increase rate was mathematically described. The results allow a better understanding of the time-course of two distinct features of bone growth: increase in size and increase in mineral content.

Absorptiometry, Photon↗

Analgesic effect of intranasal and intramuscular salmon calcitonin in post-menopausal osteoporosis: a double-blind, double-placebo study.

Different types of calcitonin (porcine, human, salmon) are used in the management of bone diseases characterized by a high bone turnover, such as post-menopausal osteoporosis and Paget's disease; recently, salmon calcitonin has become clinically available as an intranasal (i.n.) spray. An analgesic effect has also been described for calcitonins, both in experimental animals and humans, but only a few studies in humans were placebo controlled. The aim of this study was to compare the analgesic efficacy of i.n. and intramuscular (i.m.) salmon calcitonin (sCT) and of placebo in women affected by painful post-menopausal osteoporosis, in a double-blind, double-placebo trial. Twenty-eight women were randomly allocated to one of the following treatments: 1) i.n. sCT 200 U/day plus i.m. placebo; 2) i.n. placebo plus i.m. sCT 100 U/day; and 3) i.n. and i.m. placebo. Each treatment lasted four weeks, and the pain score was evaluated weekly by means of a visual analogic scale (VAS). Twenty-four women completed the trial; with i.n. sCT, the pain score decreased significantly by the second week of treatment (p < 0.05); with i.m. sCT and with placebo, the pain score decreased significantly only by the fourth week (p < 0.05), so that the final pain scores obtained with the three treatments were not different. We conclude that i.n. sCT was probably more rapid, but not more effective than i.m. sCT or placebo in decreasing pain in post-menopausal osteoporosis.

Administration, Intranasal↗

A new mathematical model to study bone turnover in growing rats.

A new mathematical model for the study of bone turnover in growing rats was developed. The model predicts a linear relationship between bone mineral content (BMC) and biochemical markers (BMK) of bone turnover assuming that rats are growing, bone turnover is profoundly affected by skeletal maturation, and resorption and formation are physiologically balanced. The model validation was performed by measuring galactosyl-hydroxylysine (GHYL) and hydroxyproline (HYP) in urines. This mathematical evidence supports our proposed use of the specific bone resorption marker GHYL to predict bone mineral content. Further studies on bone turnover will be possible by the application of the same approach.

Absorptiometry, Photon↗

Thyroid hormones and active calcium transport of inside-out red cell membrane vesicles.

Thyroid hormones may influence the active transport of Ca2+ across the cell membrane. To test the physiologic relevance of this mechanism, we used inside-out human red cell membrane vesicles as a model of the cell membrane Ca2+ pump. We monitored by spectrophotometric methods the kinetics of the uptake of Ca2+ in the presence of 10(-5)-10(-10) M thyroid hormones or their analogues. Vesicles freed of calmodulin and protein inhibitor(s) of the Ca2+ pump were also obtained. The results are as follows: (1) Thyroxine inhibits the active Ca2+ uptake; (2) this effect antagonizes that of soluble calmodulin; and (3) triiodothyronine and other analogues of the thyroid hormones are less active than thyroxine. We conclude that the thyroid hormones may influence cell Ca2+ homeostasis by direct action on the Ca2+ pump.

Adult↗

[Nutrients, anthropometric characteristics and osteoporosis in women in the recent and late postmenopausal period].

Bone mineral content (BMC) at two different radial sites (mid-diaphysis and ultra-distal epiphysis), anthropometric measurements (Body Mass Index, Lipidic Area, and Muscular Area) and nutrients intake were measured in two populations of women selected on the basis of early (< 9 years) or remote (> 15 years) menopause. The results show the presence of positive relationships between BMC, and protein and lipid intakes in the population of women in early menopause; in the other population no relationships were found. Glucid, fibre and calcium intakes were not related to the BMC of both populations. The positive relationships between BMC, and protein and lipid intakes in the population of women in early menopause is likely mediated by anthropometric characteristics as Body Mass Index and Muscular Area.

Adult↗

[Influence of nutrition, age and vitamin D status on fasting urinary excretion of calcium in postmenopausal women].

The purpose of the present study was to evaluate the influence of nutrition, age, and Vitamin D status on fasting urinary calcium (Ca) as a function of creatinine excretion (Ca/Cr) and of glomerular filtration rate (Ca/GFR) in postmenopausal women. Fasting urinary calcium, urinary Cr and GFR were measured in 18 women aged 54 to 91 years before and after six days of a calcium (400 mg/die) and sodium (100 mEq/die) restricted diet with a controlled content of proteins, lipids and glucides. Aged (over 64 y.) women having an adequate vitamin D status showed fasting urinary Ca/Cr and Ca/GFR after the controlled diet significantly lower than those showed before. It is concluded that fasting Ca/Cr and Ca/GFR may be nutrition-dependent. A revision of the significance of the fasting urinary Ca excretion is consequently suggested.

Age Factors↗

[Dual-energy X-ray absorptiometry of the vertebrae: determining which vertebrae should be scanned].

This study was aimed at verifying whether bone mineral density (BMD) and its loss with aging are different in the lumbar vertebrae and whether the region of interest--which is usually limited to the L2-L4 segment--may be extended to L1-L4. BMD was measured by means of dual-energy X-ray absorptiometry (Hologic QDR 1000) in 74 women, aged 37 to 78 years, not affected with any disease or subjected to any treatment known to interfere with bone metabolism. The relationship between age and BMD was expressed by the following equations for L1, L2, L3: BMD-L1 = 1181.68-7.85 x age, BDM-L2 = 1251.57-7.70 x age, BMD-L3 = 1231.66-6.57 x age, as shown by linear regression analysis. The behavior of the bone density of L4 with age appeared to be different and could not be described by linear regression curves and was therefore not comparable with that of the other vertebrae. BMD was different in the four lumbar vertebrae as shown by: a) the diversity of the intercept of the regression lines describing age-bone density relationships (F = 7.7, p < 0.001); b) the bone density of L1 being lower than the mean bone density of the L2-L4 region (p < 0.005); c) the bone density of L4 being higher than the mean bone density of the L1-L3 region (p < 0.001). In order to evaluate the effect of senile degenerative changes of the vertebrae on the relevant bone density, two groups of women were considered, according to age--i.e., pre- or iuxtamenopausal and late postmenopausal. It appeared that the BMD of L1 was always more correlated to the bone density of L2 and L3 than the BMD of L4. Our results suggest that L1 is homogeneous to the L2-L3 region, relative to both bone density and its loss with aging. Therefore, L1 should be included in the region of interest during the examination. Even though the bone mineral content of L4 and its loss with aging seem to be different, our results do not allow the exclusion of L4 from the scanned area.

Absorptiometry, Photon↗

Human red cell age, oxygen affinity and oxygen transport.

The [2,3-DPG]/[Hb] ratio and the P50 were found to be lower in the 10% denser (old) than in the 10% lighter (young) red blood cell (RBC) fractions (0.57 +/- 0.13 vs 0.96 +/- 0.13 and 23.02 +/- 0.85 vs 27.47 +/- 1.05 Torr, respectively, mean +/- SD, P less than 0.0005 for both, n = 6). The RBC aging processes appear thus to affect the RBC oxygen affinity. However, the [2,3-DPG] changes do not fully explain the drop of not fully explain the drop of P50 as measured at constant [H+], [CO2] and [HbCO]. It is therefore postulated that an additional factor is involved in the regulation of the oxygen affinity in the ageing RBC. The RBC density in 59 normal individuals matched for age (infants, adult, and aged) and for sex was found to be younger in adult females than in all other groups (P less than 0.0005), including an age-matched group of pregnant women. Correspondingly, the [2,3-DPG]/[Hb] ratio and the P50 are higher in adult females than in adult males (0.92 +/- 0.10 vs 0.82 +/- 0.09, P less than 0.009, and 29.03 +/- 1.07 vs 27.72 +/- 0.82 Torr, P less than 0.002, respectively). These data are evaluated in terms of the efficiency of the oxygen transport calculating the circulatory load required to transport a given amount of oxygen to the tissues. The results indicate that the lower oxygen affinity (due to the younger RBC population) in adult females partially compensates for their lower [Hb].

Adult↗

Red cell aging and active calcium transport.

The authors have investigated the relationships between the active calcium transport across the human red blood cell (RBC) membrane and the RBC aging processes in vivo and in vitro. For the study of this biological system, the authors have determined the active calcium uptake by inside-out membrane vesicles obtained from selected RBC populations. This model provided an optimal way to assess the biochemical and functional responses of the human cell to the oxidative stimulus triggered by the cellular aging processes. The activity of the calcium pump is indeed strictly correlated to the oxidative damage suffered by the RBC, being higher in the aged RBC. It appears that the main controller of the active calcium transport is the age-dependent protein inhibitor of the calcium pump.

Adult↗

The effect of in vitro and in vivo cellular aging on the active calcium transport in human inside-out red cell membrane vesicles.

Modelling of the in vivo and in vitro aging processes in the human red cell has stressed the following features of the active calcium uptake by inside-out vesicles: 1) it is higher in the outdated, in vitro aged, than in the fresh red cell (p less than 0.0005), and in the densest, in vivo aged fraction than in the lightest, young fraction (p = 0.08); 2) it increases following stimulation by excess calmodulin to values that are not significantly different; 3) it decreases to the same value in the absence of endogenous calmodulin and inhibitor, with and without exogenous calmodulin; 4) it is the target of a non-competitive inhibition, that is stronger in the fresh than in the outdated red cell. We conclude that the aging process does not involve neither membrane Ca-ATPase nor calmodulin, but rather the interaction of the calcium pump with the inhibitor of Ca-ATPase.

Biological Transport, Active↗