Bone loss physiology in critically ill patients.
Explore the source record for details and available documents.
Biomarkers↗
Biomedical subjects
G E Lester
Publications and source records attributed to G E Lester.
Isolation of intact collagen fibrils from healing ligament.
The inability to isolate intact collagen fibrils has limited the study of their growth and structure. Although intact fibrils have been isolated from echinoderms and from embryonic chick tissues, no method has previously succeeded in isolating intact collagen fibrils from a postfoetal vertebrate tissue. Having previously observed that gentamicin weakens rat tail tendon, we hypothesized that gentamicin may weaken interfibrillar bonds and that intact collagen fibrils might be isolated from tissue treated with gentamicin. In this study medial collateral knee ligaments of Sprague Dawley rats were transected and then harvested 24, 48 or 96 h postoperatively. These specimens were placed in gentamicin or phosphate-buffered saline for 72 h, vortexed for 1 h, incubated in gentamicin for an additional 24 h, and vortexed again for 1 h. Negatively stained specimens were examined with a transmission electron microscope. The phosphate-buffered saline specimens yielded only broken fibrils. The gentamicin specimens yielded both broken and intact fibrils. The latter and tapering ends and consisted of molecules orientated such that their amino termini pointed toward the tip and their carboxy termini pointed toward a short central region where the molecular polarity reversed.
Animals↗
A mathematical approach to the definition of osteoporosis.
Explore the source record for details and available documents.
Humans↗
Beta-carotene content of postharvest orange-fleshed muskmelon fruit: effect of cultivar, growing location and fruit size.
The influence of two growing locations (soil types), six fruit sizes, and two years on the postharvest Beta-carotene content of muskmelon (Cucumis melo L. var. reticulatus Naud.) fruit was studied with two different cultivars. Fully abscised commercial size fruit: 9, 12, 15, 18, 23, and 30 (fruit/0.04 M3 shipping box) had highly variable Beta-carotene contents (5.3 to 33.8 micrograms/g fresh weight) that varied by size class, soil type and cultivar. Beta-carotene content increased with fruit size up to a maximum, though fruit size continued to increase. Find sandy loam soil produced fruit with less Beta-carotene content than silty clay loam soil. The cultivar Primo contained higher Beta-carotene content levels than cultivar Cruiser. Mesocarp percent moisture content for both 'Cruiser' and 'Primo' at both locations by fruit size was not significantly correlated (r = 0.40) with Beta-carotene content. Indicating fruit cell dilution may not contribute to the differences in Beta-carotene content in different fruit size classes. Beta-carotene content of size class '18' fruit from six cultivars grown on the silty clay loam soil for two consecutive years, showed a year, and year by cultivar effect for some cultivars. Whereas, some cultivars did not differ in Beta-carotene content between the two years. This indicates a potential for controlling Beta-carotene content of muskmelon fruit at a constant, high level by careful selection of production cultivar.
Crops, Agricultural↗
Differential distribution of lumican and fibromodulin in tooth cementum.
The objectives of this study were to isolate and characterize the major proteoglycans of tooth cementum in relation to the tissue's mineralization. Cementum was collected from the root apex region of bovine molars and pulverized. It was first extracted with 6M guanidine-HCI, pH 7.4 (G-extract, mineral-unassociated) and then demineralized and extracted with 0.5M EDTA (E-extract, mineral-associated). Both extracts were applied to anion exchange and then molecular sieve chromatography to isolate proteoglycans. The fractions collected were assayed for chondroitin-(CS) and keratan sulfate (KS) containing proteoglycans using the monoclonal antibodies 2-B-6 and 5-D-4, respectively. It was found that the KS was the major glycosaminoglycan and was enriched in the G-extract fraction. The major KS fraction was then applied to 7.5% SDS-PAGE. The major broad band (69 kDa) was 5-D-4 positive in Western blot analysis and separated into two bands (46 kDa and 50 kDa) after treatment with keratanase II and endo-beta-galactosidase. These two proteins were transfered to PVDF membrane and analyzed for amino acid sequence. The results showed the major band (46 kDa) to be lumican and the minor (50 kDa) fibromodulin. In addition, based on the immunohistochemical study using a number of mono- and polyclonal antibodies including 5-D-4, anti-lumican core protein as well as anti-fibromodulin core protein antibodies, the KSPGs were found to be located almost exclusively in nonmineralized portions of cementum such as precementum and the pericementocyte area. These biochemical as well as immunohistochemical data suggest that the major KSPGs of cementum, lumican and fibromodulin, have a specific tissue distribution and may have regulatory roles in cementum mineralization.
Amino Acid Sequence↗
Effect of cefazolin and vancomycin on osteoblasts in vitro.
The effect of cefazolin and vancomycin on osteoblast-like cells was studied. Cells from the MG-63 human osteosarcoma cell line were grown in antibiotic free media and exposed to concentrations of cefazolin and vancomycin at order of magnitude intervals between 0 and 10,000 microg/ml. For cefazolin, a second interval was performed between 100 and 1000 microg/ml to define toxic levels more accurately. Cell number and 3H-thymidine incorporation at 0, 24, and 72 hours were determined. The results of this study show that local levels of vancomycin of 1000 microg/ml and less have little or no effect on osteoblast replication, and concentrations of 10,000 microg/ml cause cell death. Concentrations of cefazolin of 100 microg/ml and less have little or no effect on osteoblast replication, 200 microg/ml significantly decrease cell replication, and 10,000 microg/ml cause cell death. The authors conclude that vancomycin is less toxic than is cefazolin to osteoblasts at higher concentrations and may be a better antibiotic for local administration in the treatment of similarly sensitive bacterial infections.
Anti-Bacterial Agents↗
EDTA-insoluble, calcium-binding proteoglycan in bovine bone.
A calcium ion precipitable, trypsin-generated proteoglycan fragment has been isolated from the demineralized, EDTA-insoluble matrices of bone. The demineralized matrix was completely digested with trypsin, increasing concentrations of CaCl2 were added to the supernatant, and the resulting precipitates were analyzed. The amount of precipitate gradually increased with higher concentrations of calcium and was reversibly solubilized by EDTA. After molecular sieve and anion exchange chromatography, a proteoglycan-containing peak was obtained. Immunochemical analysis showed that this peak contained chondroitin 4-sulfate and possibly keratan sulfate. Amino acid analysis showed that this proteoglycan contained high amounts of aspartic acid/asparagine (Asx), serine (Ser), glutamic acid/glutamine (Glx), proline (Pro), and glycine (Gly); however, it contained little leucine (Leu) which suggests that it is not a member of the leucine-rich small proteoglycan family. In addition, significant amounts of phosphoserine (P-Ser) and hydroxyproline (Hyp) were identified in hydrolysates of this fraction. A single band (M(r) 59 kDa) was obtained on SDS-PAGE that stained with Stains-all but not with Coomassie Brilliant Blue R-250. If bone powder was trypsinized prior to demineralization, this proteoglycan-containing fraction was not liberated. Collectively, these results indicate that a proteoglycan occurs in the demineralized matrix that is precipitated with CaCl2 and is closely associated with both mineral and collagen matrices. Such a molecule might facilitate the structural network for the induction of mineralization in bone.
Amino Acids↗
Bone toxicity of locally applied aminoglycosides.
The effect of tobramycin on osteoblasts was studied. Osteoblast-like cells from the MG-63 human osteosarcoma cell line were grown in antibiotic-free media and exposed to concentrations of tobramycin: first at order of magnitude intervals between 0 and 10,000 micrograms/ml, and then at closer intervals between 100 and 1,000 micrograms/ml to more accurately define toxic levels. Cell number and 3H-thymidine incorporation at 0, 24 and 72 h were determined. Similarly, primary cultures of rat osteoblasts were exposed to the same concentrations of tobramycin to confirm the validity of the MG-63 cell line as a model for nontransformed cells. The results of this study demonstrate that local levels of tobramycin of < or = 200 micrograms/ml have little or no effect on osteoblast replication. Concentrations of 400 micrograms/ml significantly decreased cell replication, and concentrations of 10,000 micrograms/ml caused cell death. Results obtained with primary rat osteoblast cultures were similar to those obtained from the MG-63 cultures at the tested tobramycin concentrations.
Animals↗
Increase in serum calcium after administration of PTH.
Explore the source record for details and available documents.
Animals↗
Update on the use of distal radial bone density measurements in prediction of hip and Colles' fracture risk.
A controversy has developed around the question as to whether bone density values from the distal radius can be used to accurately predict risk of future fractures. To address this question, two separate studies were undertaken: (a) Bone density was measured in 460 healthy ambulatory women living in retirement centers in the state of North Carolina; 83% of these women were followed for up to 60 months for occurrence of minimal trauma hip and wrist fractures. Thirty-one minimal trauma fractures were reported in our study population, representing 8% of those followed. The fracture incidence density rate showed a close inverse relationship with incremental changes in bone density at the distal site. Twenty-eight of the 31 fractures were reported in women with bone density values below the 325-mg/cm2 "at risk" value. (b) Bone density values of the distal radius and the lumbar spine from 360 women (aged 18-85 years) from the Chapel Hill area were used to analyze the error in predicting individual spinal density from the distal radial density. Although the overall correlation was high (r = 0.67) and the confidence intervals were narrow, the prediction intervals were quite wide. Thus, prediction of an individual value of spine density from the distal radius density would result in a value with a range too wide to be clinically useful. We conclude that single-photon absorptiometry appears to be a useful tool for screening normal populations of asymptomatic women for prediction of hip or Colles' fracture risk even though it has limited usefulness in prediction of spinal fracture risk or individual values for spinal density.
Aged↗
Regulation of serum 1,25-dihydroxyvitamin D3 in lactating rats.
To characterize further the mechanism(s) underlying the increased serum 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] concentration associated with lactation in the rat, we examined hormone biosynthesis [i.e., renal 25-hydroxyvitamin D-1 alpha-hydroxylase (1 alpha-hydroxylase) activity] and hormone disappearance in groups of lactating Holtzman rats and age- and sex-matched nonlactating controls. 1 alpha-Hydroxylase activity was significantly greater in kidneys from lactating rats (4.0 +/- 0.42 fmol.mg-1.min-1) on a basal diet than in those from nonmated females (1.4 +/- 0.08 fmol.mg-1.min-1), an increment sufficient to account for the observed fourfold elevation of 1,25(OH)2D3 in the dams. The increase occurs despite the lower serum 1,25(OH)2D3 levels in lactating than in nonlactating rats at 12 and 24 h after a bolus injection of 1,25(OH)2D3 (2 ng/g body wt). Elevation of serum 1,25(OH)2D3 is not a requisite consequence of lactation, however, because dams receiving supplemental calcium from food (1.6%) and water (0.3%) exhibited no increase of either serum 1,25(OH)2D3 or 1 alpha-hydroxylase activity compared with controls. In contrast, lactating rats that received a diet with only 0.1% calcium had 5-fold higher serum 1,25(OH)2D3 levels and 20-fold higher 1 alpha-hydroxylase activity than nonlactating rats on the same diet. We conclude that other factors in conjunction with lactation, but not the lactating state per se, promote the changes in 1,25(OH)2D3 metabolism observed.
25-Hydroxyvitamin D3 1-alpha-Hydroxylase↗
Cross-sectional moment of inertia of the distal radius from absorptiometric data.
Explore the source record for details and available documents.
Absorptiometry, Photon↗
Stress shielding effect of rigid internal fixation plates on mandibular bone grafts. A photon absorption densitometry and quantitative computerized tomographic evaluation.
The use of rigid internal fixation reconstruction plates in combination with bone grafts provides many advantages in mandibular reconstruction. These advantages include ease of adaptability at the time of surgery as well as immediate postoperative function for the patient. Disadvantages may include a "stress shielding" or disuse osteoporosis effect on the underlying bone graft as a result of the rigidity of the reconstruction plate. This study evaluated the stress shielding effects on bone grafts in primate mandibles. Bilateral bone grafts stabilized with rigid fixation plates were performed in 4 Macaca fascicularis monkeys. Three months after the bone grafting procedure one plate was removed from each monkey. The monkeys were then killed 1, 4, 6 and 8 months after plate removal. Photon absorption densitometry and computerized tomography were used to evaluate stress shielding. The results of this study indicate that the stress shielding effect occurs if the plate is not removed from the area of bone grafting postoperatively.
Absorptiometry, Photon↗
Histomorphometric evaluation of stress shielding in mandibular continuity defects treated with rigid fixation plates and bone grafts.
Histomorphometric techniques were used to evaluate the stress shielding effect in bilateral bone grafts in 4 Macaca fascicularis monkeys. Bilateral continuity defects were created and grafted by replacing the resected portion of the mandible and iliac crest bone into the defect area. Both sides were plated with rigid internal fixation plates. Three months after bone grafting, the plate was removed from one side while the other plate was left in place. The animals were sacrificed at 1, 4, 6, and 8 months after plate removal and the grafted areas removed for histomorphometric analysis. This study documents decreased bone volume as well as smaller interlabel width on the plated side, suggesting a stress shielding effect as a result of the rigid internal fixation plate.
Animals↗
Cholecalciferol and placental calcium transport.
Transplacental movement of calcium from mother to fetus is essential for normal fetal development. In most species, fetal plasma calcium levels are higher than maternal levels at term. The role of cholecalciferol metabolites, with specific emphasis on 1,25-dihydroxycholecalciferol (1,25(OH)2D), in placental calcium transport and maintenance of the fetomaternal gradient has been extensively investigated. In rats, there is not an absolute demand for 1,25(OH)2D for maintenance of fetal calcium homeostasis in utero, even though it is essential for maintenance of maternal plasma calcium levels. However, in sheep, the absence of 1,25(OH)2D results in disruption of both maternal and fetal calcium homeostasis. It is known that rat and human placentas contain specific cytosolic binding proteins for 1,25(OH)2D that are similar to the well-characterized intestinal receptor. Two calcium-binding proteins (CaBP) have been detected in rat and human placentas: a protein immunologically identical to the vitamin D-dependent CaBP and a calcium-dependent ATPase. The levels of CaBP in rat placenta have been shown to increase in response to exogenously administered 1,25(OH)2D but cannot be obliterated with maternal vitamin D deficiency. No relationship has been shown between 1,25(OH)2D and placental Ca-ATPase in any species. Thus, the mechanism of action of 1,25(OH)2D in maintenance of the transplacental calcium gradient in sheep is unknown. In the pregnant rat (and perhaps human), 1,25(OH)2D is a critical factor in the maintenance of sufficient maternal calcium for transport to the fetus and may play a role in normal skeletal development of the neonate.
24,25-Dihydroxyvitamin D 3↗
In vivo bone mineral analysis throughout skeletal growth in rats: differences due to sex or vitamin D deficiency.
This study reports a new technique for the measurement of bone mineral content (BMC) in live rats. A single photon absorptiometric instrument has been adapted for rapid, reproducible measurement of BMC. Four bone sites were selected for use, based on ease of positioning and reproducibility of measurement; these were as follows: proximal femur, midfemur, proximal seventh caudal vertebra, and midseventh caudal vertebra. Significant increases were detected when BMC was measured at biweekly intervals from weaning to 18 weeks of age in normal male rats. Comparison of body growth and changes in BMC of male and female rats with age showed that body weight gain of female rats slowed earlier than that of male rats whereas BMC increased at similar rates in both sexes. Vitamin D deprivation from day 24 of life resulted in decreased BMC at all four measurement sites compared with such measurements in normal control rats. Differences were detectable after 8 weeks of age and occurred despite the maintenance of serum calcium levels within normal range and only slight reduction in body weights of vitamin D deprived rats. These studies demonstrate that single photon absorptiometry can be used to monitor changes in BMC in live rats on a routine basis without harm to the animals. Changes in BMC such as those due to growth or vitamin D deprivation can easily be quantitated using this technique.
Animals↗
Formation of 19-nor-10-keto-25-hydroxyvitamin D3 in cultured mammalian cells.
Cultured dog kidney cells convert 25-hydroxyvitamin D3 into more polar metabolites during in vitro incubations. One elutes with 1,25-dihydroxyvitamin D3 from high pressure liquid chromatographic silica columns with hexane:isopropanol (9:1), but can be separated from 1,25-dihydroxyvitamin D3 by elution with dichloromethane:isopropanol (95:5). This peak has been isolated, purified and identified by mass spectral analysis to be 19-nor-10-keto-25-hydroxyvitamin D3.
Animals↗
Fluctuation of serum concentration of 1,25-dihydroxyvitamin D3 during the menstrual cycle.
The serum concentrations of total calcium and 1,25-dihydroxyvitamin D3 were measured in seven women during a menstrual cycle. A rise in the serum level of 1,25-dihydroxyvitamin D3 was observed on day 15 of the cycle without a detectable change in the serum calcium level. The level of 1,25-dihydroxyvitamin D3 on day 15 was double the value at days 1 and 8 of the cycle. The peak of 1,25-dihydroxyvitamin D3 on day 15 was not observed in five women ingesting oral contraceptives, suggesting that ovulation or its hormonal control was involved in the peak on day 15. A small but significant fall in the serum calcium concentration occurred in the women ingesting oral contraceptives. In view of these findings the serum concentrations of 1,25-dihydroxyvitamin D in women with functioning ovaries must be interpreted in the context of the stage of the menstrual cycle when the blood sample was obtained.
Adolescent↗