bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of Osteocalcin Expression in Transgenic Mice Reveals a Species Difference in Vitamin D Regulation of Mouse and Human Osteocalcin Genes
Authors: Clemens Thomas L. , Tang Hui , Maeda Shigeto , Kesterson Robert A. , Demayo Francesco , Pike J. Wesley , Gundberg Caren M.
Published: April 2006

A line of transgenic mice expressing a human osteocalcin genomic fragment (hOClocus) and a murine MC3T3-E1 cell line containing a stably integrated human osteocalcin promoter construct have been developed to characterize the osteogenic and hormonal regulation of human osteocalcin in vivo and in vitro. In this study, we used these models to demonstrate a species difference in the regulation of the mouse and human osteocalcin genes by vitamin D. Repeated administration of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) to mice carrying the hOClocus transgene resulted in striking increases in serum human osteocalcin, whereas serum mouse osteocalcin levels were unchanged after 24 h and only modestly increased 48 h after the second dose of hormone. 1,25(OH)2D3 increased human calvarial mRNA expression by 1.8-fold and slightly decreased mouse osteocalcin mRNA levels by approximately 1.2-fold. Furthermore, treatment of primary calvarial osteoblasts from these mice with 1,25(OH)2D3 increased human osteocalcin production but inhibited mouse osteocalcin protein accumulation. To investigate further the mechanism for the apparent species difference in vitamin D3 induction of mouse and human osteocalcin, we examined the effect of 1,25(OH)2D3 in an MC3T3-E1 cell line (MC4) containing a stably integrated 3900 bp osteocalcin promoter-luciferase construct. Treatment of MC4 cells with ascorbic acid resulted in parallel increases of the endogenous mouse osteocalcin protein and luciferase reporter activity over a 12-day period. Continuous exposure of MC4 cells to 1,25(OH)2D3 resulted in time-and dose-dependent increases in the activity of the phOC3900 luciferase construct. By contrast, the hormone had no effect on mouse osteocalcin protein concentrations and inhibited its induction by ascorbic acid. However, when cells were treated acutely with 1,25(OH)2D3 at later times during growth in ascorbic acid, the induction of mouse osteocalcin protein was only partially inhibited. In conclusion, our results indicate that common osteogenic signals regulate both mouse and human osteocalcin gene expression, but the mouse gene is resistant to induction by vitamin D. This species difference in vitamin D regulation of osteocalcin appears to result from the failure of 1,25(OH)2D3 to transcriptionally activate the mouse osteocalcin gene.
Total Body Bone Mineral Density in Young Children: Influence of Head Bone Mineral Density
Authors: Taylor Arlene , Konrad Patricia T. , Norman Michael E. , Harcke H. Theodore
Published: April 2006

Dual-energy X-ray absorptiometry (DXA) with its short scan time, low radiation dose, and high precision and accuracy have made this technique particularly suitable for measuring total body bone mineral density (TBMD) in children. Other published reports have related TBMD to age in children 2–18 years of age. However, in young normal children aged 2–9 years (51 girls, 43 boys), we found that regression equations for TBMD with age as the predictor did not explain enough of the variance to warrant their use for predicting TBMD (adjusted R2 0.47, females; 0.41, males). Subtotal BMD (TBMD − head BMD) is predicted better by age because of a possibly invalid adult algorithm for head BMD (adjusted R2 0.73, females; 0.71, males).
The Effect of Aging on Bone Formation in Porous Hydroxyapatite: Biochemical and Histological Analysis
Authors: Inoue K. , Ohgushi H. , Yoshikawa T. , Okumura M. , Sempuku T. , Tamai S. , Dohi Y.
Published: April 2006

The effect of aging on osteoblastic differentiation of marrow stromal stem cells was examined. Porous hydroxyapatite (HA) disks were soaked in cells suspensions of bone marrow cells from young (8 weeks) and old rats (60 weeks) and then implanted subcutaneously in syngeneic young and old rats. The bone marrow/HA composites were harvested 8 weeks later, and the contents of bone Gla protein (BGP) and alkaline phosphatase (ALP) activity in them were determined. Histologically, bone formation could be detected in all the composites in young recipient rats; however, some old bone marrow/HA composites in old recipients did not show bone formation and the bone volume in the young bone marrow/HA composites was greater than in the old bone marrow/HA composites. The ratios of ALP activities of young bone marrow/HA composites to old bone marrow/HA composites in young and old recipients were about five times and four times, respectively. The ratios of BGP contents of young bone marrow/HA to old bone marrow/HA composite in young and old recipients were about nine and eight times, respectively. The results suggest that the decreased bone formation observed in old bone marrow cells was due to a smaller population of stromal cells and/or decreased capacity of differentiation of stromal stem cells into osteogenic cells.
The Vitamin D Receptor Gene Start Codon Polymorphism: A Functional Analysis of FokI Variants
Authors: Gross Coleman , Krishnan Aruna V. , Malloy Peter J. , Eccleshall T. Ross , Zhao Xiao-Yan , Feldman David
Published: April 2006

The vitamin D receptor (VDR) gene contains a start codon polymorphism (SCP) which is three codons upstream of a second start site (ATG). The SCP genotype can be determined with the restriction enzyme FokI, where “f” indicates the presence of the restriction site and the first ATG, while “F” indicates its absence. Recent evidence suggests that the ff genotype is correlated with lower bone mineral density (BMD) in some populations. The SCP results in alternate VDRs that differ structurally, with the F variant (F-VDR) being three amino acids shorter than the f variant (f-VDR). To determine whether there are functional differences between the f-VDR and the F-VDR, we studied the two VDR forms expressed in COS-7 cells. The proteins were distinguishable from one another on Western blots by their different mobilities, confirming the larger size of f-VDR. Ligand binding studies showed no significant differences between the affinities of the two VDR forms for [3H]-1,25-dihydroxyvitamin D3 ([3H]-1,25(OH)2D3) (Kd = 131 ± 78 pM, f-VDR; Kd = 237 ± 190 pM, F-VDR; p = 0.24); however, a 2-fold difference in affinity can not be discriminated by this method. There were no differences in the abilities of the two receptor forms to bind DNA as determined by electrophoretic mobility shift assays. The ability of the two VDR forms to transactivate target genes was investigated using three different vitamin D responsive luciferase reporter constructs: 24-hydroxylase, osteocalcin, and osteopontin. In these transactivation experiments, 1,25(OH)2D3 dose-response (0.1–10 nM) curves revealed that the ED50 values for transactivation were indistinguishable between the two VDR forms. Additionally, cultured human fibroblasts with FF,Ff, and ff genotypes had similar sensitivity to 1,25(OH)2D3 with respect to the induction of 24-hydroxylase mRNA. In summary, we were unable to detect significant differences in ligand affinity, DNA binding, or transactivation activity between f-VDR and F-VDR forms. We must emphasize, however, that the sensitivity of the methods used limits our ability to detect minor differences in VDR affinity and function. In conclusion, we cannot define a mechanism whereby the SCP in the VDR might contribute to population differences in BMD.
Responses of Trabecular and Cortical Bone Turnover and Bone Mass and Strength to Bisphosphonate YH529 in Ovariohysterectomized Beagles with Calcium Restriction
Authors: Yoshida Yasushi , Moriya Atsuko , Kitamura Kazuyuki , Inazu Mizuho , Okimoto Nobukazu , Okazaki Yuichi , Nakamura Toshitaka
Published: April 2006

Thirty-six beagles, 18 months of age, underwent ovariohysterectomy (OHX) or a sham operation. Sham-operated animals were given a diet with standard calcium (1.4%) (group 1, n = 6) or a restricted calcium diet (0.14%) (group 2, n = 6). The OHX animals were given the restricted calcium diet and YH529 orally with respective daily doses of 0, 0.02, 0.1, and 0.5 mg/kg of body weight (groups 3–6, n = 6 each) for 12 months. At the end of this period, the lumbar bone mineral densities (BMDs) in groups 2 and 3 and the load values for group 3 were significantly smaller than those for group 1. The midfemur BMD did not differ among the groups. The urinary deoxypyridinoline (U-Dpy) and bone formation rates (BFR/BS, BFR/BV) in groups 2 and 3 and the osteonal BFR/BS and trabecular osteoclast number (Oc.N/BS) in group 3 were significantly larger than the respective values for group 1. However, these parameters did not significantly differ between groups 2 and 3. The serum osteocalcin (OC) level, wall thickness (W.Th), and mineral apposition rate values for group 3 were significantly larger than those for group 2. In group 2, the trabecular activation frequency (Ac.F) increased by 3.11 times, and the percent values of the number of labeled osteons (L-Ot.N/T-Ot.N, %) in the tibia by 3.28 times over those for group 1. In group 3, the Ac.F increased by 3.20 times and the number of labeled osteons by 3.77 times over those for group 1. In groups 4–6, the U-Dpy and Oc.N/BS values were smaller, but their OC levels did not significantly differ from the level for group 3. The lumbar BMD, the load, and W.Th were dose-dependently significantly larger than those for group 3. The Ac.F values were significantly smaller, and the respective value in groups 4–6 was 67.9, 25.5, and 10.2% of that in group 3. The BMDs of the midfemur in groups 4–6 were significantly larger than those in group 3, but the ultimate load values did not significantly differ. The L-Ot.N/T-Ot.N values were also significantly smaller, and the respective value in groups 4–6 was 82.0, 48.5, and 55.2% of that in group 3. The tibial endocortical and periosteal BFR/BSs did not differ significantly. These data demonstrate that the effects of OHX on bone mass and turnover were small in the beagles fed a restricted calcium diet. YH529 maintained the mass and strength of the lumbar bone by reducing the bone resorption. The cortical bone appeared to be less sensitive to the agent than the trabecular bone in this animal model.
Osteoclast Differentiation Requires Ascorbic Acid
Authors: Ragab Ashraf A. , Lavish Sandra A. , Banks Michael A. , Goldberg Victor M. , Greenfield Edward M.
Published: April 2006

Osteoclast differentiation assays are usually conducted in alpha minimal essential medium (α-MEM). We reasoned that determining which components of this media are critical for osteoclast differentiation might provide insight into the mechanisms that regulate osteoclast differentiation. This study demonstrates that ascorbic acid is the crucial component of α-MEM that stimulates differentiation of murine osteoclasts in cocultures with murine mesenchymal support cells. Thus, supplementation with ascorbic acid allows osteoclast differentiation to occur in basal MEM media as well as in RPMI-1640 and basal media Eagle (BME) media. The conclusion that osteoclast differentiation is stimulated by ascorbic acid was obtained whether osteoclast differentiation was induced by 1,25-dihydroxyvitamin D3 or parathyroid hormone, whether ST2 or CIMC-2 cells were used as mesenchymal support cells, and whether osteoclast precursors were obtained from spleen or bone marrow. Time course studies revealed that although ascorbic acid only modestly increases the rate at which osteoclast precursors begin to express tartrate-resistant acid phosphatase, it strongly increases the rate at which precursors fuse into mature, multinucleated cells. Moreover, ascorbic acid strongly increases the life span of both osteoclasts and their precursors. The increases in precursor formation, fusion, and life span induced by ascorbic acid are together responsible for the stimulation of osteoclast differentiation by ascorbic acid. Given the known effects of ascorbic acid on differentiation of mesenchymal cells, it may stimulate osteoclast differentiation indirectly by regulating the differentiation state of the mesenchymal cells that support osteoclast differentiation.
Gender Differences in Insulin-like Growth Factor and Bone Mineral Density Association in Old Age: The Rancho Bernardo Study
Authors: Barrett-Connor Elizabeth , Goodman-Gruen Deborah
Published: April 2006

Insulin-like growth factor-I (IGF-I) clearly plays a role in bone metabolism and maintenance, as evidenced by in vitro and animal studies. In clinical studies, the age-related decrease in IGF-I parallels the age-related decrease in bone mineral density (BMD), but several age-adjusted cross-sectional studies show no consistent association of IGF-I and BMD. We report here a cross-sectional study of serum IGF-I and BMD levels in 483 men and 455 postmenopausal women not using estrogen; subjects were 55 years of age and older, community-dwelling, ambulatory, and unselected for bone density. IGF-I was measured by a highly specific radioimmunoassay. BMD was measured at the lumbar spine and hip using dual-energy X-ray absorptiometry. Men had higher IGF-I and BMD levels than women. In age-adjusted and age-stratified models, IGF-I was associated with BMD only in women (test for interaction, p < 0.0001). Gender differences persisted in gender-specific multiple regression analyses adjusted for age, body mass index, thiazide diuretic use, current smoking, alcohol intake, physical activity, and weight change; IGF-I was significantly associated with BMD at the spine (p = 0.0001) and hip (p = 0.02) in women, but not in men (p's > 0.6). Circulating estradiol levels were not associated with IGF-I levels in either gender; testosterone was inversely associated with IGF-I and only in men. This striking gender difference has not been described previously. Its etiology is unknown. The answer could lead to improved understanding of gender differences in osteoporosis and in response to treatment with IGF-I or growth hormone.
Cathepsin K Knockout Mice Develop Osteopetrosis Due to a Deficit in Matrix Degradation but Not Demineralization
Authors: Gowen Maxine , Lazner Francesca , Dodds Robert , Kapadia Rasesh , Feild John , Tavaria Michael , Bertoncello Ivan , Drake Fred , Zavarselk Silva , Tellis Irene , Hertzog Paul , Debouck Christine , Kola Ismail
Published: April 2006

Cathepsin K is a cysteine protease expressed predominantly in osteoclasts. Activated cathepsin K cleaves key bone matrix proteins and is believed to play an important role in degrading the organic phase of bone during bone resorption. Mutations in the human cathepsin K gene have been demonstrated to be associated with a rare skeletal dysplasia, pycnodysostosis. The degree of functional activity of the mutated forms of cathepsin K in these individuals has not been elucidated, but is predicted to be low or absent. To study the role of cathepsin K in bone resorption, we have generated mice deficient in the cathepsin K gene. Histologic and radiographic analysis of the mice revealed osteopetrosis of the long bones and vertebrae, and abnormal joint morphology. X-ray microcomputerized tomography images allowed quantitation of the increase in bone volume, trabecular thickness, and trabecular number in both the primary spongiosa and the metaphysis of the proximal tibiae. Not all bones were similarly affected. Chondrocyte differentiation was normal. The mice also had abnormalities in hematopoietic compartments, particularly decreased bone marrow cellularity and splenomegaly. The heterozygous animals appeared normal. Close histologic examination of bone histology revealed fully differentiated osteoclasts apposed to small regions of demineralized bone. This strongly suggests that cathepsin K–deficient osteoclasts are capable of demineralizing the extracellular matrix but are unable to adequately remove the demineralized bone. This is entirely consistent with the proposed function of cathepsin K as a matrix-degrading proteinase in bone resorption.
The Epidemiology of Paget's Disease in Britain: Is the Prevalence Decreasing?
Authors: Prof. Cooper Cyrus , Schafheutle Karen , Dennison Elaine , Kellingray Samantha , Guyer Peter , Barker David
Published: April 2006

To estimate changes in the age- and gender-specific prevalence of Paget's disease in Britain, we performed a radiographic survey of the disorder in 10 British centers, using sampling and radiographic methods identical to a study performed in 1974. In each center, a sample of abdominal radiographs of people aged 55 years and over was taken from stored films within the radiology department of the principal general hospital. The radiographs were identified by screening radiographic records over the period 1993–1995. Any abdominal radiograph in a subject aged 55 years and over which included the entire pelvis, sacrum, femoral heads, and all lumbar vertebrae was studied. The radiographs were evaluated by a trained observer and the consultant radiologist who participated in the original 1974 survey. Nine thousand eight hundred and twenty-eight radiographs (4625 men, 5203 women) were assessed in the 10 towns. The overall age/gender standardized prevalence rate was 2%, with a male/female ratio of 1.6. Prevalence increased steeply with age among men and women, rising to 6.9% of men and 5.8% of women aged 85 years and over. The prevalence of Paget's disease in the 10 towns in 1994 was only 40% of that observed during the 1974 study. The decline in prevalence was apparent in all 10 centers, but was most marked in those with high rates in the original study. This survey of Paget's disease in 10 British towns suggests a prevalence of 2.5% among men and 1.6% among women aged 55 years and over. Age-adjusted prevalence rates declined steeply between 1974 and 1994. These declines suggest an environmental contribution to the etiology of this disorder that requires further investigation.
Sympathectomy, Which Induces Membranous Bone Remodeling, Has No Effect on Endochondral Long Bone Remodeling In Vivo
Authors: Sherman Bret E. , Chole Richard A.
Published: April 2006

Sympathectomy has been shown to induce resorption within the membranous middle ear bone of gerbils. It is unknown whether sympathectomy exerts a similar effect on endochondral long bone. In the present study, guanethidine sulfate (GS) and 6-hydroxydopamine (HDA) were administered to gerbils to induce sympathectomy. One week later, samples of middle ear bulla bone and radial long bone were harvested and assessed for osteoclastic activity. Histomorphometric analysis showed both pharmacologic sympathectomy with GS and chemical sympathectomy with HDA significantly increased the osteoclast counts and osteoclast surfaces of bulla bone samples but not radial long bone samples, respectively. In contrast, HDA but not GS increased the osteoclast profile area of both long bone and membranous bone samples when compared with vehicle-treated controls. Sympathectomy, induced both chemically and pharmacologically, thus has been shown to increase resorption in membranous bone but not endochondral long bone in the gerbilline model.
Osteoblastic 11β-Hydroxysteroid Dehydrogenase Type 1 Activity Increases With Age and Glucocorticoid Exposure
Authors: Cooper Mark S. , Rabbitt Elizabeth H. , Goddard Philippa E. , Bartlett William A. , Hewison Martin , Stewart Paul M.
Published: April 2006

The risk of glucocorticoid-induced osteoporosis increases substantially with age but there is considerable individual variation. In recent studies we have shown that the effects of glucocorticoids on bone are dependent on autocrine actions of the enzyme 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1); expression of 11β-HSD1 in osteoblasts (OBs) facilitates local synthesis of active glucocorticoids with consequent effects on osteoblastic proliferation and differentiation. Using primary cultures of human OBs, we have now characterized the age-specific variation in osteoblastic 11β-HSD1 and defined enzyme kinetics and regulation using natural and therapeutic glucocorticoids. 11β-HSD1 reductase activity (cortisone to cortisol conversion) was recognized in all OB cultures and correlated positively with age (r = 0.58 with all cultures, p < 0.01, and n = 18; r = 0.87 with calcaneal-derived cultures, p < 0.001, and n = 14). Glucocorticoid treatment caused a time- and dose-dependent increase in 11β-HSD1 activity over control (e.g., dexamethasone [DEX; 1 μM], 2.6-fold ± 0.5 (mean ± SE), p < 0.001, and n = 16; cortisol (100 nM), 1.7-fold ± 0.1, p < 0.05, and n = 14). Similar increases in 11β-HSD1 mRNA expression were indicated using real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) analyses (3.5-fold with DEX, p < 0.01; 2.5-fold with cortisol, p < 0.05). The capacity of 11β-HSD1 to metabolize the synthetic glucocorticoids prednisone and prednisolone was investigated in human OBs (hOBs) and fetal kidney-293 cells stably transfected with human 11β-HSD1 cDNA. Transfected cells and hOBs were able to interconvert prednisone and prednisolone with reaction kinetics indistinguishable from those for cortisone and cortisol. To assess the in vivo availability of substrates for osteoblastic 11β-HSD1, plasma cortisone and prednisone levels were measured in normal males before and after oral prednisolone (5 mg). The 9:00 a.m. serum cortisone levels were 110 ± 5 nmol/liter and prednisone levels peaked at 78 ± 23 nmol/liter 120 minutes after administration of prednisolone. Thus, therapeutic use of steroids increases substrate availability for 11β-HSD1 in bone. These studies indicate that activation of glucocorticoids at an autocrine level within bone is likely to play an important role in the age-related decrease in bone formation and increased risk of glucocorticoid-induced osteoporosis.
Onion and a Mixture of Vegetables, Salads, and Herbs Affect Bone Resorption in the Rat by a Mechanism Independent of Their Base Excess
Authors: Mühlbauer Roman C. , Lozano Annemarie , Reinli Andreas
Published: April 2006

Prevention of low bone mass is important to reduce the incidence of osteoporotic fractures. In man, the consumption of fruits and vegetables is associated with greater bone mineral density (BMD), an effect that is claimed to be caused by their base excess buffering metabolic acid, thought to dissolve bone. We showed previously that in the rat the consumption of several vegetables, salads, and herbs inhibits bone resorption and that onion increases bone mass. In this study we show that, although the intake of onion is associated with a decrease in urinary noncarbonic acid excretion and a concomitant inhibition of bone resorption of similar magnitude, the two findings are not causally related. Onion retains its bone resorption inhibitory activity in the rat even when added to a vegetarian diet with typical base excess. Onion and a mixture of vegetables, salads, and herbs retain their inhibitory activity even when metabolic acid is buffered with potassium citrate. In addition, neither the pH nor the potassium content of individual ashed vegetables, salads, and herbs correlates with inhibition of bone resorption. The effect of vegetables, salads, and herbs, which inhibit bone resorption in the rat, therefore is not mediated by their base excess but possibly by a pharmacologically active compound(s).
Tower Climbing Exercise Started 3 Months After Ovariectomy Recovers Bone Strength of the Femur and Lumbar Vertebrae in Aged Osteopenic Rats
Authors: Notomi Takuya , Okimoto Nobukazu , Okazaki Yuichi , Nakamura Toshitaka , Suzuki Masashige
Published: April 2006

To determine both the preventive and recovery effects of tower climbing exercise on mass, strength, and local turnover of bone in ovariectomized (OVX) rats, we carried out two experiments. In experiment I, 60 Sprague-Dawley rats, 12 months of age, were assigned to four groups: a Baseline Control, Sham-Operated Sedentary, OVX-Sedentary and OVX-Exercise rats. Rats voluntarily climbed a 200-cm tower to drink water from a bottle set at the top. At 3 months, OVX elevated both the femoral cortex and lumbar trabecular turnover, leading to a reduction in bone mass and strength. However, in OVX-Exercise rats, those values were maintained at the same level as in the Sham-Sedentary rats. Thus, the climbing exercise, started after 3 days of OVX, prevented OVX-induced cortical and trabecular bone loss by depressing turnover elevation. After confirming the preventive effect, we evaluated the recovery effect of exercise. In experiment II, 90 Sprague-Dawley rats, 12 months of age, were assigned to six groups: a Baseline control, two groups of Sham-Operated Sedentary and OVX-Sedentary, and OVX-Exercise rats. The exercise started 3 months after the OVX operation. At 3 months, OVX increased the trabecular bone formation rate and osteoclast surface, leading to a decrease in compressive strength. In the midfemur, the cross-sectional area, moment of inertia, and bending load values decreased. At 6 months, in the OVX-Exercise rats, the parameters of breaking load in both the lumbar and midfemur, lumbar bone mass, and the total cross-sectional area recovered to the same levels as those in the Sham-Sedentary rats. However, the cortical bone area did not recover. Periosteal bone formation increased, while endosteal bone formation decreased. These results showed that the climbing exercise had both a preventive and recovery effect on bone strength in OVX rats. In the mid-femur, effects on bone formation were site-specific, and the cross-sectional morphology was improved without an increase in cortical bone area, supporting cortical drift by mechanical stimulation.
The Ability of Statins to Inhibit Bone Resorption Is Directly Related to Their Inhibitory Effect on HMG-CoA Reductase Activity
Authors: Staal Ada , Frith Julie C , French Michele H , Swartz Joann , Güngör Timur , Harrity Thomas W , Tamasi Joseph , Rogers Michael J , Feyen Jean HM
Published: April 2006

Statins, which are inhibitors of 3-hydroxy-3-glutaryl-coenzyme A (HMG-CoA) reductase, decrease the hepatic biosynthesis of cholesterol by blocking the mevalonate pathway. Nitrogen-containing bisphosphonate drugs also inhibit the mevalonate pathway, preventing the production of the isoprenoids, which consequently results in the inhibition of osteoclast formation and osteoclast function. Therefore, we hypothesized that statins could affect bone metabolism in vivo through effects on osteoclastic bone resorption. In vitro, cerivastatin inhibited the parathyroid hormone (PTH)-stimulated bone resorption. Using a panel of 40 statin analogs, which showed variable effects on HMG-CoA reductase activity, we found that the ability of compounds to inhibit bone resorption is directly related to HMG-CoA reductase activity. However, in the thyro-parathyrodectomy (TPTX) model for bone resorption in the rat in vivo, cerivastatin did not prevent experimentally induced increases in bone resorption. The lack of effect of cerivastatin in this model is not related to a limited penetration of the target tissue (bone marrow), because a significant effect on HMG-CoA reductase activity was demonstrated in the total rat bone marrow cell extracts of rats posttreatment in vivo. Furthermore, cerivastatin inhibited protein prenylation in osteoclasts isolated from the rabbit bone marrow of rabbits after treatment in vivo. In contrast to other studies, none of the statins tested showed anabolic effects in parietal bone explant cultures. Taken together, we conclude that statins inhibit bone resorption in vitro, which correlates directly with the potency of the compounds for inhibition of HMG-CoA reductase activity. However, cerivastatin does not affect bone resorption in the rat TPTX model in vivo.
BMP-2 Controls Alkaline Phosphatase Expression and Osteoblast Mineralization by a Wnt Autocrine Loop
Authors: Rawadi Georges , Vayssière Béatrice , Dunn Fred , Baron Roland , Roman-Roman Sergio
Published: April 2006

Wnt/β-catenin signaling has recently been suggested to be involved in bone biology. The precise role of this cascade in osteoblast differentiation was examined. We show that a Wnt autocrine loop mediates the induction of alkaline phosphatase and mineralization by BMP-2 in pre-osteoblastic cells.

Introduction: Loss of function of LRP5 leads to osteoporosis (OPPG syndrome), and a specific point mutation in this same receptor results in high bone mass (HBM). Because LRP5 acts as a coreceptor for Wnt proteins, these findings suggest a crucial role for Wnt signaling in bone biology.

Materials and Methods: We have investigated the involvement of the Wnt/LRP5 cascade in osteoblast function by using the pluripotent mesenchymal cell lines C3H10T1/2, C2C12, and ST2 and the osteoblast cell line MC3T3-E1. Transfection experiments were carried out with a number of elements of the Wnt/LRP5 pathway. Measuring osteoblast and adipocyte differentiation markers addressed the effect of this cascade on osteoblast differentiation.

Results: In mesenchymal cells, only Wnt's capable of stabilizing β-catenin induced the expression of alkaline phosphatase (ALP). Wnt3a-mediated ALP induction was inhibited by overexpression of either Xdd1, dickkopf 1 (dkk1), or LRP5ΔC, indicating that canonical β-catenin signaling is responsible for this activity. The use of Noggin, a bone morphogenic protein (BMP) inhibitor, or cyclopamine, a Hedgehog inhibitor, revealed that the induction of ALP by Wnt is independent of these morphogenetic proteins and does not require de novo protein synthesis. In contrast, blocking Wnt/LRP5 signaling or protein synthesis inhibited the ability of both BMP-2 and Shh to induce ALP in mesenchymal cells. Moreover, BMP-2 enhanced Wnt1 and Wnt3a expression in our cells. In MC3T3-E1 cells, where endogenous ALP levels are maximal, antagonizing the Wnt/LRP5 pathway led to a decrease of ALP activity. In addition, overexpression of dkk1 reduced extracellular matrix mineralization in a BMP-2-dependent assay.

Conclusions: Our data strongly suggest that the capacity of BMP-2 and Shh to induce ALP relies on Wnt expression and the Wnt/LRP5 signaling cascade. Moreover the effects of BMP-2 on extracellular matrix mineralization by osteoblasts are mediated, at least in part, by the induction of a Wnt autocrine/paracrine loop. These results may help to explain the phenotype of OPPG patients and HBM.
Telomerase Accelerates Osteogenesis of Bone Marrow Stromal Stem Cells by Upregulation of CBFA1, Osterix, and Osteocalcin
Authors: Gronthos Stan , Chen Shaoqiong , Wang Cun-Yu , Robey Pamela G , Shi Songtao
Published: April 2006

Telomerase activity can prevent telomere shortening and replicative senescence in human somatic cells. We and others have previously demonstrated that forced expression of telomerase in human bone marrow stromal stem cells (BMSSCs) was able to extend their life-span and enhance their bone-forming capability, without inducing malignant transformation. In this study, we determined that telomerase was able to accelerate calcium accumulation of human BMSSCs under osteogenic inductive conditions. Similarly, xenogeneic transplantation of telomerase-expressing BMSSCs (BMSSC-Ts) yielded ectopic bone formation at 2 weeks post-transplantation, 2–4 weeks earlier than typically seen with BMSSCs transfected with empty vector (BMSSC-Cs). Low-density DNA array analysis revealed that telomerase activity increases the expression of G1 regulating genes including cyclin D3, cyclin E1, E2F-4, and DP2, associated with hyperphosphorylation of retinoblastoma (pRb), leading to the extended proliferative capacity of BMSSC-Ts. Importantly, BMSSC-T transplants showed a higher number of human osteogenic cells at 8 weeks post transplantation compared with the BMSSC-C transplants, coupled with a significantly increased osteogenic capacity. One possible mechanism leading to accelerated osteogenesis by BMSSC-Ts may be attributed, at least in part, to the upregulation of the important osteogenic genes such as CBFA1, osterix, and osteocalcin in vitro. Taken together, these findings show that telomerase can accelerate cell cycle progression from G1-to-S phase and enhance osteogenic differentiation of BMSSCs, because of the upregulation of CBFA1, osterix, and osteocalcin.
Intracrystalline Proteins and Urolithiasis: A Synchrotron X-ray Diffraction Study of Calcium Oxalate Monohydrate
Authors: Fleming David E , Van Riessen Arie , Chauvet Magali C , Grover Phulwinder K , Hunter Brett , Van Bronswijk Wilhelm , Ryall Rosemary L
Published: April 2006

The existence of intracrystalline proteins and amino acids in calcium oxalate monohydrate was demonstrated by X-ray synchrotron diffraction studies. Their presence has implications for the destruction of calcium oxalate crystals formed in the urinary tract and the prevention of kidney stones.

Introduction: Although proteins are present in human kidney stones, their role in stone pathogenesis remains unknown. This investigation aimed to characterize the nature of the relationship between the organic and mineral phases in calcium oxalate monohydrate (COM) crystals grown in human urine and in aqueous solutions of proteins and amino acids to clarify the function of proteins in urolithiasis.

Methods: COM crystals were grown in human urine and in aqueous solutions containing either human prothrombin (PT), Tamm-Horsfall glycoprotein (THG), aspartic acid (Asp), aspartic acid dimer (AspAsp), glutamic acid (Glu), glutamic acid dimer (GluGlu), or γ-carboxyglutamic acid (Gla). Controls consisted of COM crystals precipitated from pure inorganic solutions or from human urine that had been ultrafiltered to remove macromolecules. Synchrotron X-ray diffraction with Rietveld whole-pattern peak fitting and profile analysis was used to determine nonuniform crystal strain and crystallite size in polycrystalline samples.

Results: Crystals precipitated from ultrafiltered urine had lower nonuniform strain than those grown in urine or in aqueous PT solution. Nonuniform strain was much lower in crystals grown in distilled water or in the presence of THG. For the amino acids, the highest nonuniform strain was exhibited by crystals grown in Gla solution, followed by Glu. Crystallite size was inversely related to nonuniform strain, with the effect being significantly less for amino acids than for macromolecules.

Conclusions: Selected proteins and amino acids associated with COM crystals are intracrystalline. Although their incorporation into the mineral bulk would be expected to affect the rate of crystal growth, they also have the potential to influence the phagocytosis and intracellular destruction of any crystals nucleated and trapped within the renal collecting system. Crystals impregnated with protein would be more susceptible to digestion by cellular proteases, which would provide access to the crystal core, thereby facilitating further proteolytic degradation and mineral dissolution. We therefore propose that intracrystalline proteins may constitute a natural form of defense against renal stone formation.
Neuroprotection by Transgenic Expression of Glucose-6-Phosphate Dehydrogenase in Dopaminergic Nigrostriatal Neurons of Mice
Authors: Mejías Rebeca , Villadiego Javier , Pintado C. Oscar , Vime Pablo J. , Gao Lin , Toledo-Aral Juan J. , Echevarría Miriam , López-Barneo José
Published: April 2006

Oxidative damage to dopaminergic nigrostriatal (DNS) neurons plays a central role in the pathogenesis of Parkinson's disease (PD). Glucose-6-phosphate dehydrogenase (G6PD) is a key cytoprotective enzyme that provides NADPH, the major source of the reducing equivalents of a cell. Mutations of this enzyme are the most common enzymopathies worldwide. We have studiedin vivothe role of G6PD overexpressed specifically in the DNS pathway and show that the increase of G6PD activity in the soma and axon terminals of DNS neurons, separately from other neurons or glial cells, protects them from parkinsonism. Analysis of DNS neurons by histological, neurochemical, and functional methods showed that even a moderate increase of G6PD activity rendered transgenic mice more resistant than control littermates to the toxic effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). The neuroprotective action of G6PD was also observed in aged animals despite that they had a greater susceptibility to MPTP. Therefore, overexpression of G6PD in dopaminergic neurons or pharmacological activation of the native enzyme should be considered as potential therapeutic strategies to PD.
Neocortical Network ActivityIn VivoIs Generated through a Dynamic Balance of Excitation and Inhibition
Authors: Haider Bilal , Duque Alvaro , Hasenstaub Andrea R. , McCormick David A.
Published: April 2006

The recurrent excitatory and inhibitory connections between and within layers of the cerebral cortex are fundamental to the operation of local cortical circuits. Models of cortical function often assume that recurrent excitation and inhibition are balanced, and we recently demonstrated that spontaneous network activityin vitrocontains a precise balance of excitation and inhibition; however, the existence of a balance between excitation and inhibition in the intact and spontaneously active cerebral cortex has not been directly tested. We examined this hypothesis in the prefrontal cortexin vivo, during the slow (<1 Hz) oscillation in ketamine–xylazine-anesthetized ferrets. We measured persistent network activity (Up states) with extracellular multiple unit and local field potential recording, while simultaneously recording synaptic currents in nearby cells. We determined the reversal potential and conductance change over time during Up states and found that the body of Up state activity exhibited a steady reversal potential (−37 mV on average) for hundreds of milliseconds, even during substantial (21 nS on average) changes in membrane conductance. Furthermore, we found that both the initial and final segments of the Up state were characterized by significantly more depolarized reversal potentials and concomitant increases in excitatory conductance, compared with the stable middle portions of Up states. This ongoing temporal evolution between excitation and inhibition, which exhibits remarkable proportionality within and across neurons in active local networks, may allow for rapid transitions between relatively stable network states, permitting the modulation of neuronal responsiveness in a behaviorally relevant manner.
CA150 Expression Delays Striatal Cell Death in Overexpression and Knock-In Conditions for Mutant Huntingtin Neurotoxicity
Authors: Arango Margarita , Holbert Sébastien , Zala Dania , Brouillet Emmanuel , Pearson James , Régulier Etienne , Thakur Ashwani Kumar , Aebischer Patrick , Wetzel Ronald , Déglon Nicole , Néri Christian
Published: April 2006

Transcriptional dysregulation caused by expanded polyglutamines (polyGlns) in huntingtin (htt) may be central to cell-autonomous mechanisms for neuronal cell death in Huntington's disease (HD) pathogenesis. We hypothesized that these mechanisms may involve the dysfunction of the transcriptional regulator CA150, a putative modifier of onset age in HD, because it binds to htt and accumulates in an HD grade-dependent manner in striatal and cortical neurons. Consistently, we report herein that CA150 expression rescues striatal cell death in lentiviral overexpression (rats) and knock-in (mouse cells) conditions for mutant htt neurotoxicity. In both systems, rescue was dependent on the (Gln-Ala)38repeat normally found in CA150. We excluded the possibility that rescue may be caused by the (Gln-Ala)38repeat interacting with polyGlns and, by doing so, blocking mutant htt toxicity. In contrast, we found the (Gln-Ala)38repeat is required for the nuclear restriction of exogenous CA150, suggesting that rescue requires nuclear CA150. Additionally, we found the (Gln-Ala)38repeat was dispensable for CA150 transcriptional repression ability, suggesting further that CA150 localization is critical to rescue. Finally, rescue was associated with increased neuritic aggregation, with no reduction of nuclear inclusions, suggesting the solubilization and nuclear export of mutant htt. Together, our data indicate that mutant htt may induce CA150 dysfunction in striatal neurons and suggest that the restoration of nuclear protein cooperativity may be neuroprotective.
Trajektorienplanung für die Steuerung gewisser linearer Systeme mit verteilten Parametern (Trajectory Planning for the Control of some Linear Distributed Parameter Systems)
Authors: Rudolph Joachim , Woittennek Frank
Published: April 2006

Der Entwurf von Steuerungen und die Trajektorienplanung für lineare Systeme mit örtlich verteilten Parametern und konzentrierten Stelleingriffen können in gewissen Fällen auf der Basis von Potenzreihendarstellungen erfolgen. Es wird eine Methode vorgestellt, mit der diese Reihendarstellungen systematisch auch für solche partiellen Differentialgleichungen hergeleitet werden können, deren Koeffizienten von der Ortsvariablen abhängen. Als Anwendungsbeispiel für den Fall ortsabhängiger Koeffizienten wird eine ringförmige, dünne elastische Platte betrachtet, für die eine Steuerung entworfen wird, die der Platte ein umlaufendes Biegeprofil aufprägt.
Eating Ivy
Authors: harper lisa
Published: April 2006

This essay is both celebration and portrait of the late Barton Rouse and his influential career as chef at the Terrace Club of Princeton University during the writer's undergraduate career. In the late 1980s and early 1990's Barton Rouse transformed a dilapidated building into a culinary mecca. He planned inventive daily menus (chrysanthemum soup; broiled tuna with morel sauce; White Trash Night) and hosted extravagant special events: an anti-Valentine's day dinner, for instance, which featured Blackened Rib Steaks and Catfish Fillets, Black fettuccine with sour cream and lox sauce and red and black caviar, bleeding hearts of beet salad, brandied cherry ambrosia, and mocha espresso cheesecake. The man and his food broke boundaries between high culture and low, good taste and bad, east and west, rural and urban, adult and child. In the process of cooking for his members, Rouse taught hundreds of young people how to eat, but also that cooking was a labor of love and a genuine aesthetic pursuit. His humor, whimsy, and inventive extravagance left a legacy which links inextricably food and politics to our fundamental way of being in the world.
Q
Authors: tribur zoe
Published: April 2006

Q is a Taiwanese word for a food texture that forms a distinctive part of the island's cuisine. Squishy and yielding, Q makes a surprisingly refreshing accompaniment to a wide range of flavors, but it is not exactly a familiar, or even welcome, mouth feel for someone, such as the author, whose previous gastronomic preferences were defined by the typical standards of mainstream America. However, approached with an open mind, Q can prove to be quite addictive, as well as a "gateway drug" to other unfamiliar and exquisite culinary experiences. For the author, Q has certainly been an acquired taste. Embracing it, along with other strange and unexpected experiences, has enriched and opened the author’s mind considerably.
Ethnolinguistic Chicago: Language and Literacy in the City's Neighborhoods
Authors: Barton Ellen
Published: April 2006

Ethnolinguistic Chicago: Language and Literacy in the City's Neighborhoods By Marcia Farr. ed. Mahwah, NJ. Lawrence Erlbaum. 2004.
African American English in the Diaspora
Authors: Green Lisa
Published: April 2006

African American English in the Diaspora: By Shana Poplack and Sali Tagliamonte. Malden, MA. Blackwell. 2001.
Association Analysis as a Strategy for Improvement of Quantitative Traits in Plants
Authors: Breseghello Flavio , Sorrells Mark E.
Published: April 2006

Association analysis is a method potentially useful for detection of marker‐trait associations based on linkage disequilibrium, but little information is available on the application of this technique to plant breeding populations. With appropriate statistical methods, valid association analysis can be done in plant breeding populations; however, the most significant marker may not be closest to the functional gene. Bias can arise from (i) covariance among markers and QTL, frequently related to population structure or intense selection and (ii) differences in initial frequencies of marker alleles in the population, such that exclusive alleles tend to be in higher association. The potentials and limitations of germplasm bank collections, synthetic populations, and elite germplasm are compared, as experimental materials for association analysis integrated with plant breeding practice. Synthetics offer a favorable balance of power and precision for association analysis and would allow mapping of quantitative traits with increasing resolution through cycles of intermating. A model to describe the association between markers and genes as conditional probabilities in synthetic populations under recurrent selection is proposed, which can be computed on the basis of assumptions related to the history of the population. This model is useful for predicting the potential of different populations for association analysis and forecasting the response to marker‐assisted selection.
Wayang and Ludruk: Polarities in Java
Authors: Hatley Barbara
Published: April 2006

In contemporary Western society, theatre has become primarily a source of entertainment and sometimes of social criticism; its older function of articulating and instilling prevailing values is now performed by the mass media and specialized educational and propaganda institutions. In Indonesia, however, and elsewhere in Asia where mass media and educational facilities are less developed and private reading is not a widely established habit, popular drama still transmits sociopolitical ideals between the court (or city) and the countryside, and reinforces religious tradition. While some of its communicative role is now shared with movies and other new media, live theatre is still a vital and popular teacher.

In Java the Wayang Kulit, or shadow-puppet theatre, epitomizes the traditional Javanese world view; it has functioned as an art form, a religious rite, and a medium of instruction for more than a thousand years.
The Grand Union
Authors: Paxton Steve
Published: April 2006

The Grand Union was formed in the Fall of 1970. Founding members of this anarchistic democratic theatre collective were: Becky Arnold, Trisha Brown, Dong, Douglas Dunn, David Gordon, Nancy Green, Barbara Lloyd, Steve Paxton and Yvonne Rainer. Characteristic of the Union, there are nine different answers to how and when it began. The lives of the members have been intertwined for anywhere from five to ten years. In 1970, the members of Yvonne Rainer's Continuous Project Altered Daily (CPAD) conceived of an organization in which they might share choreography. Three other choreographer/performers of similar mind were invited to join. This was the beginning of the organization but not of the concept.
The Classification of Circus Techniques
Authors: Burgess Hovey
Published: April 2006

In writing on theatre, various authors have pointed out that circus-type performances have always been more prevalent than performances of literary drama; that circus-type performance has been a deep secondary tradition of the theatre from which the legitimate stage has repeatedly drawn new strength and vitality; and that the circus is looked to by the public “for violent satisfactions, the intentions of which do not deceive them.” I use the term “circus-type” to emphasize an important distinction between the very ancient “circus-type” skills of contortionists, equilibrists, tumblers, jugglers, clowns, etc., and the modern form of “circus,” even though the latter is the most obviously significant depository of the former.

The modem circus is by no means a direct descendant of the ancient Roman circus. Roman circuses, such as Circus Maximus, were architectural structures designed primarily for chariot races.