Phosphorus utilization in chicks fed hydrated sodium calcium aluminosilicate.

Two experiments were conducted to evaluate the effects of hydrated Na Ca aluminosilicate (HSCAS) on P utilization of young broiler chicks. Phosphorus-deficient corn-soybean meal diets containing .36% (.134% available) P and 1% Ca were fortified with 0, .05 and .10% P provided as KH2PO4 (22.8% P) or feed-grade dicalcium phosphate (18.9% P). Diets were available ad libitum to chicks receiving 0, .50 or 1.0% HSCAS during the period 8 to 22 d posthatching. Weight of tibia ash, percentage of tibia ash and bone-breaking force were regressed on supplemental P intake to assess P utilization in the absence or presence of HSCAS. Tibia parameters (ash weight, percentage of ash and bone-breaking force) responded linearly (P less than .01) to P supplementation. Regardless of P source (KH2PO4 or dicalcium phosphate), .5% or 1.0% HSCAS had no effect (P greater than .1) on P utilization. With no supplemental P, tibia parameters also were not affected (P greater than .05) by HSCAS. Phosphorus utilization from dicalcium phosphate was estimated to be 87% as efficient as that observed for KH2PO4. The results suggest that dietary HSCAS does not impair utilization of either phytate or inorganic P.

Separation of non-collagenous proteins of bone formed in titanium implants: experimental study in the rabbit tibia, using a bone harvest chamber.

The non-collagenous proteins of mature cortical rabbit bone and 3 wk old bone specimens formed in titanium implant bone harvest chambers, were separated by means of fine pressure liquid chromatography. It was found that a sufficient amount of bone could be obtained from five bone harvest chambers in five rabbits during a period of 15 wk. A different protein chromatographic pattern was found in bone grown in bone harvest chambers as compared to normal cortical bone from the same region. This indicates a difference in bone quality and shows that this implant design can be used for biochemical analysis of bone formed adjacent to a biomaterial.

[Effect of immobilization on the physical properties of cancellous bone: an experimental study].

40 adult new Zealand rabbits were divided into 6 groups, including a control group, which was free from immobilization. The left hind limb of each animal in the experimental groups was immobilized with the knee in extension by plaster cast for different period of time: 2,4,6,8 and 10 weeks for different group. The cancellous bone of upper tibia of each animal was tested systematically. The bone gravity, pointpressing strength, ash ratio and mineral content were measured and compared group by group to investigate their changes under different conditions. Following immobilization, the quantitative reduction of each parameter varied with the length of immobilization, marked in the first 6 weeks but gradually slowed down. It was most obvious in pointpressing strength, then mineral content, and the least the apparent gravity, denoting that the bone strength was most sensitive to immobilization. The reduction of mineral content exceeded that gravity, indicating the main loss was the former. Loss of point-pressing strength and mineral content was more prominent in the posterior part of the plateau, the weight-bearing area, than at the anterior and intercondylar parts. So was the metaphyseal area as compared with the epiphyseal area. In general, changes in bone gravity was directly in proportion with that of bone strength.

Effect of feeding rabbits on naturally moulded and mycotoxin-contaminated diet.

For studying the effects of naturally moulded diet on mature female-rabbits, Buskat rabbits were fed for 6 weeks on a naturally moulded diet comparing with the other similar group of animals which was fed also on the same ingredients in the first diet, except that, they were sound (not moulded apparently). The results which were obtained from this study due to the feeding on the moulded diet were as follows: 1. Decreasing the feed intake significantly (P less than 5%) and increasing somewhat the water consumption, but the water/feed ratio increased significantly (P less than 5%). 2. Reduction of the digestibility of most nutrients, especially of the crude fibre (P less than 1%) and crude protein (P less than or equal to 5%). 3. Changing in the relative weights of different organs particularly the significant (P less than 5%) decrease in weight of the liver and spleen as well as the significant (P less than 5%) increase in weight of empty stomach and female genital tract. 4. The most important haematological changes were the reduction in cholesterol content and the elevation of phospholipid concentration, thus the cholesterol/phospholipid ratio decreased by the rate of 40.7%. Calcium/phosphorus ratio increased by the rate of 9.5% as a result of increasing calcium content in the control serum. 5. Chemical analyses revealed a significant (P less than 1%) reduction in ash content of the liver and magnesium content of the tibia bone, a slight decrease in liver content of iron and ether extract, and significant (P less than 5%) rise in femoral muscle content of the ether extract.

Effect of occupational lead exposure on serum 1,25-dihydroxyvitamin D levels.

The effects of lead exposure on serum 1,25-dihydroxyvitamin D levels and calcium homeostasis have been studied in 63 males occupationally exposed to the metal in the UK. The exposure indices used were blood lead, reflecting short-term exposure, and an in vivo X-ray fluorescence measurement of tibia lead which reflects cumulative lead exposure. Serum 1,25-dihydroxyvitamin D levels were higher than those in a referent population, who were non-occupationally exposed to lead, and were correlated with both blood lead and tibia lead. Multiple regression analysis suggested that blood lead was the variable responsible for the increase in serum 1,25-dihydroxyvitamin D. There were no other abnormalities in calcium metabolism associated with the degree of lead exposure.

The effect of high-dose salmon calcitonin on bone mineral metabolism in the normal rat.

The paucity of information on the effect of long-term high-dose salmon calcitonin administration on normal bone mineral metabolism and histology prompted an investigation of the influence of high-dose synthetic calcitonin in the rat. Serum ionized calcium, osteocalcin or BGP (bone gla protein), and immunoreactive PTH were measured serially during calcitonin administration and bone histomorphometry analyzed at 6 weeks (after sacrifice). Daily injections of salmon calcitonin, 0.4 IU/100 g (group B) and 2 IU/100 g (group C), resulted in significant hypocalcemia at 4 hours for both experimental groups (P less than 0.004). Serum iPTH was significantly higher over the study period for both groups administered calcitonin. Serum BGP levels were significantly lower than controls during the study in group C (P less than 0.002) and to a lesser extent in group B (P less than 0.05). In group C, bone histomorphometry revealed increased resorption (osteoclast count), decreased trabecular bone volume, and decreased double-labeled tetracycline surface (bone formation). In group B an increase in osteoclast count but no alteration in bone formation was observed. To assess the role of PTH in the above findings, high-dose calcitonin was administered to parathyroidectomized rats. All of the above changes in bone histomorphometry were not observed in this group of animals. In conclusion, high doses of calcitonin promote hypocalcemia, secondary hyperparathyroidism, and osteoclastosis in the normal rat in a dose-dependent manner with very high-dose calcitonin impairing bone formation.

Beta-carotene supplementation of turkey diets varying in fat and retinol.

1. The effects of supplementary beta-carotene (BC) at 0, 1.5, 15, 150, 1500 or 15,000 mg/kg diet on performance, incidence of leg problems, tibia ash, and plasma and liver concentration of BC and retinol of poults to 4 weeks of age were studied. 2. Body weight and food intake increased with dietary BC supplementation. Incidence of leg problems was not affected but tibia ash increased with BC supplementation. Plasma and liver BC and liver retinol increased with dietary BC but plasma retinol was not affected. 3. The effects of 0, 1.5, 15 or 150 mg BC/kg in diets containing either 30 or 60 g added fat/kg and either 1.2 or 12.0 mg retinol/kg on the above properties were studied in poults to 4 weeks of age. 4. Performance was not significantly affected by dietary fat. Efficiency of utilisation of food was depressed by the higher concentration of retinol. At 3 weeks of age birds given BC had fewer leg problems and higher tibia ash values than the control birds. Liver BC and retinol and plasma BC concentration increased with dietary BC. By 4 weeks, plasma and liver BC was higher and liver retinol was lower for birds given 1.2 compared with those given 12.0 mg/kg of retinol.

In vivo measurements of bone lead--a comparison of two x-ray fluorescence techniques used at three different bone sites.

In vivo bone lead measurements have been made on a group of about 120 people, most of whom were lead exposed workers. Two different x-ray fluorescence (XRF) techniques were used to make measurements at three bone sites. Finger lead was measured using 57Co sources, and lead measurements were made in both tibia and calcaneus with a technique based on 109Cd sources. The results of the bone lead measurements correlated strongly with each other and with the index of cumulative exposure, thus confirming the value and reliability of these in vivo measurements as a tool in the study of chronic lead exposure. Measurement precision, +/- 1 standard deviation, was highest for tibia +/- 7.4 micrograms (g bone mineral)-1, +/- 16.6 micrograms (g bone mineral)-1 for the calcaneus and lowest for phalangeal lead +/- 25.0 micrograms (g bone mineral)-1. Maximum absorbed doses to the skin were comparable for all three measurements (1-3 mGy). The mean whole body dose equivalents were all low, but that for the finger measurement, 0.1 microSv, was significantly less than for the calcaneus and tibia measurements 3-5 microSv.

Dual photon absorptiometry of the proximal tibia.

Bone mineral content (BMC) and bone mineral density (BMD) of the proximal tibia were determined by dual photon absorptiometry on 44 women, aged 23-87 years. The area of the tibia measured was a 2.01 cm region immediately distal to the medial and lateral tuberosities. Values of BMC ranged between 5.09 and 14.57 g and BMD between 0.380 and 1.180 g/cm2. Both tibial BMC and BMD declined with age and tibial BMD was significantly correlated with lumbar spine (r = 0.70), femoral neck (r = 0.73), and femoral trochanter (r = 0.74). However, the large standard errors of estimate (SEE) (0.08-0.14 g/cm2) do not allow for reliable prediction in an individual of other skeletal sites by the tibia. Repeated measurements demonstrated that dual photon absorptiometry of the proximal tibia is a reliable measurement and may be a useful tool in the monitoring of therapeutic or intervention modalities in those individuals with skeletal diseases in whom measurement of the lumbar spine or proximal femur may not be possible.

Clinical examination of weanling rats with early zinc deficiency.

In order to gain experience about the detection of adverse effects during a scientific procedure, we carried out a clinical examination of rats with zinc deficiency. In weanling rats fed a zinc-deficient diet (30 mumol zinc/kg) for 10 days, the mean tibial concentration of zinc was reduced by 53% and body weight gain by 73% when compared with rats fed a diet containing an adequate amount of zinc (150 mumol zinc/kg). In a small open field on day 9 of the experiment, the deficient rats more frequently displayed the posture standing upright with elevated heels. On day 10 of the experiment a clinical examination was carried out at random and 'blind' by three independent assessors. Out of 20 variables scored quantitatively on each individual animal, only body size differed between normal and deficient rats. Other classical signs of zinc deficiency, such as alopecia, dermatitis and diarrhoea, were not detected. It is concluded that in this rat model of zinc deficiency, no evidence for extreme discomfort can be demonstrated.

Synthesis and secretion of Ch 21 protein in embryonic chick skeletal tissues.

We reported the identification, purification and characterization of a low molecular weight protein (Ch 21) expressed in vitro by differentiating chondrocytes at a late stage of development and observed in vivo in the growth plate region of the long bones at the border between hypertrophic cartilage and newly formed bone (Descalzi Cancedda, F., P. Manduca, C. Tacchetti, P. Fossa, R. Quarto, R. Cancedda, J. Cell Biol. 107, 2455-2463 (1988]. In this article, the synthesis and location of Ch 21 protein in the chick embryo tibia at late stage of development were further investigated. Ch 21 was observed in the cartilage matrix surrounding marrow cavities and in the prearticular outer layer by immunolocalization. In addition, the timing of Ch 21 appearance during the tibia development and its distribution in the growth plate region was better defined. We first observed presence of Ch 21 in the perichondral mid-diaphyseal sleeve of 7-day-old tibia. Ch 21 antibodies stained also the newly formed bone. Synthesis and secretion in the culture medium of Ch 21 protein was observed when bone fragments or cultured osteoblasts isolated from 19-day-old embryo tibiae were labeled in vitro. A search for the presence of Ch 21 in the chick embryo sternum was performed. The synthesis of Ch 21, both in the presumptive calcification cranial portion and in the permanent cartilaginous caudal portion of the sternum, was shown by metabolic labeling of tissue slices.(ABSTRACT TRUNCATED AT 250 WORDS)

Bone disorder in cardiomyopathic hamsters.

Bones of cardiomyopathic hamsters (UM-X7.1 Syrian hamsters), at 5, 10 and 20 weeks of age, were compared chemically and histomorphologically with those of normal Syrian hamsters. Femurs of UM-X7.1 hamsters were significantly shorter than those of normal hamsters, and the mean dry weight, mean volume, mean ash weight per unit bone volume and mean ash as a percentage of dry weight of femurs were all significantly less in UM-X7.1 hamsters. The bone disorder preceded the myocardial calcium precipitation and myocardial hypertrophy in the cardiomyopathic hamsters. In addition, the percentage of cortical area measured on the cross-section of tibia and the appositional rate of bone minerals, determined by a tetracycline labelling technique, were also lower in the UM-X7.1 hamsters. These findings suggest that the bone disorder was associated with decreased bone formation in the UM-X7.1 Syrian hamsters.

Effect of dietary sodium zeolite A and excess calcium on growth and tibia calcium and phosphorus concentration in uninfected and Eimeria acervulina-infected chicks.

Two identical trials were conducted with 5 to 14-day-old broiler chicks. Sodium zeolite A (NZA, 0 and .75%) and Ca (1.0 and 1.5%) were fed to both uninfected and Eimeria acervulina-infected chicks resulting in a 2 x 2 x 2 factorial arrangement of treatments. Coccidial infection reduced weight gain and gain:feed (P less than .01), and tibia ash, Ca, and Ca:P ratio (P less than .05). Excess dietary Ca reduced (P less than .05) weight gain and tibia ash in uninfected chicks but had no effect (P greater than .10) in coccidiosis-infected chicks (Ca x coccidiosis interaction, P less than .05). Addition of NZA to diets with excess Ca further decreased (P less than .05) weight gain and tibia ash in both uninfected and E. acervulina-infected chicks. Tibia Ca, as a percentage of dry fat-free tibia, was reduced (P less than .05) by the addition of NZA. This effect was not observed when tibia Ca was expressed as a percentage of ash. These results suggest that NZA may exacerbate the adverse effects of excess dietary Ca.

Quantifying manganese in lymphocytes to assess manganese nutritional status.

To clarify whether manganese nutritional status is better reflected by the manganese concentration in lymphocytes or in whole blood, we injected manganese solutions intravenously into manganese-deficient rats and determined manganese concentrations in lymphocytes, whole blood, and various tissues. The manganese concentrations in lymphocytes and tissues, but not in whole blood, were significantly less in manganese-deficient rats than in normal rats. These low values could be prevented by intravenous injection of manganese in a dose-dependent manner. These results suggest that, for assessment of manganese nutritional status, measurement of manganese in lymphocytes is better than that in whole blood.

1,25-Dihydroxyvitamin D3 prevents the decrease of bone mineral appositional rate in rats with inflammation-mediated osteopenia (IMO).

We have studied the effects of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on bone mass and bone mineral appositional rate in intact rats and rats with inflammation-mediated osteopenia (IMO), where osteoblast number and mineral appositional rate are decreased. 1,25(OH)2D3 prevents IMO-specific bone loss when given in a daily dose of 25 ng per rat, but does not when given in higher doses. The hormone was effective, when given over the complete duration of the experiment (21 days), but not when given over shorter time periods (7 and 14 days, respectively). 1,25(OH)2D3 prevents IMO-dependent reduction in mineral appositional rate and leads to an only moderate increase in intact rats. We conclude, that 1,25(OH)2D3 is more effective in stimulating mineral appositional rate in rats with IMO where mineral apposition is impaired.

Possibility of fat addition in the rabbit diets.

Baladi rabbits were fed on five-similar-experimental diets, except the replacement for starch in the 1st diet, cattle tallow in the 2nd, cotton seed oil in the 3rd, and hydrogenated palm oil in the 4th instead of 2% more wheat bran in the 5th (control) diet. All other husbandry conditions were the same for all groups of animals during the experimental period of 7 weeks. The cattle tallow in the second diet caused significant increase of feed intake, growth rate, relative weights of kidneys, lungs and heart and calcium of the tibia bone. This diet had tendency to diminish significantly blood contents of total nitrogen and cholesterol as well as vitamin A in the liver and tibia contents of silica, phosphorus and magnesium. Diet number 3 included cottonseed oil lowered blood contents of glucose, phosphorus, cholesterol and enzyme activity of glutamate oxaloacetate transaminase in the serum and specific gravity of tibia bone. On the other hand, it elevated significantly (P less than or equal to 0.01) stored vitamin A in the liver than on all other experimental diets. Feeding rabbits on diet including hydrogenated palm oil subsided liver contents of dry matter, ash and vitamin A and raised ether extract of the liver significantly. It reduced also dry matter content of the femoral muscle. Substitution for starch (instead of 2% of the diet fats or bran) increased blood content of haemoglobin and haematocrit (insignificant) but values of glucose and phosphorus as well as liver content of dry matter, content of femoral muscle of dry matter and ether extract and content of tibia bone of silica and phosphorus were significantly higher than the other experimental diets. It decreased relative weights of different organs (significantly) and liver contents of ether extract and vitamin A (insignificantly) than on control diet. It could be said that the addition of cattle tallow and cottonseed oil would be recommended to be included in rabbit diets after more studies to determine the effects of the different animal-vegetable mixtures of fats, the best ratio between the two sources of fats, the interrelationships between that mixtures and the energy of the diet, the dietary protein level, the rabbit breed and their weights and aim of the production under the seasonal variation of the weather.

Lower vertebral bone density in male long distance runners.

Bone density in 13 male long distance runners (28.7 +/- 1.2 yr, 67.6 +/- 2.0 kg) and 11 male nonrunners (26.8 +/- 1.3 yr, 71.0 +/- 2.2 kg) was compared. Bone was measured at the lumbar spine and mid-tibia using dual photon absorptiometry and at the mid-radius using single photon absorptiometry. Runners (mean weekly training 92.2 +/- 6.3 km) had significantly lower (P less than 0.05) vertebral bone mineral density (1.12 +/- 0.03 g.cm-2) than nonrunners (1.24 +/- 0.04 g.cm-2). Tibial and radial bone mineral density did not differ between the groups. Daily calcium intake for runners (1,373 +/- 486 mg) and nonrunners (1,267 +/- 236 mg) exceeded the RDA. The results of this study suggest that long distance running may lead to decreased vertebral bone mineral density. The hormonal changes that occur with endurance training may contribute to this decrease.

First bone formation and the dissection of an osteogenic lineage in the embryonic chick tibia is revealed by monoclonal antibodies against osteoblasts.

Detailed studies of the origin and differentiation of osteogenic cells can be facilitated by cell-specific markers. To this end, we immunized mice with a heterogeneous population of chick embryonic bone cells and subsequently generated and selected for monoclonal antibodies against cell surface determinants. Supernatants from growing hybridoma colonies were screened immunohistochemically against frozen sections of embryonic stage 35 (day 9.5) chick tibiae. Three cell lines, SB-1, SB-2, and SB-3, which each secrete a different antibody against osteogenic cells, have been successfully cloned, stabilized, and immortalized. Antibody SB-1 reacts with a family of cells in embryonic bone, liver, kidney, and intestine, which are identically stained by the histochemical stain for alkaline phosphatase. The SB-2 antigen is present only on osteoblasts, while the SB-3 antigen is expressed on the surface of osteoblasts, ependymal cells and ventricular myoblasts. Studies on the developmental progression of osteoblasts in the embryonic tibia indicate that the determinants recognized by SB-1, SB-2, and SB-3 are temporally coupled to the appearance of the pre-osteoblast marker alkaline phosphatase. Detailed morphologic analyses reveal that SB-1 reacts with a large family of osteogenic cells residing between the surface of newly formed bone and the overlying periosteal osteoprogenitor cells. By contrast, SB-2 and SB-3 appear to react with those mature osteoblasts involved in the secretion and mineralization of osteoid. Cells which are imprisoned within bone matrix (osteocytes) of the developing tibia are not recognized by these antibodies, but are immunostained by an osteocyte-specific monoclonal antibody which does not react with SB-1, SB-2 or SB-3 positive cells. The results reported here suggest the existence of an osteogenic cell lineage which is characterized by a series of discrete cell states prior to the overt expression of the Secretory Osteoblastic phenotype. We propose that the emergence and abatement of phenotypically distinct osteogenic cell surface antigens follows a precise spatial and temporal sequence which reflects the position of cells within the osteogenic lineage.