Masaharu Tsutsumi

Multiple vertebral fractures are associated with refractory reflux esophagitis in postmenopausal women

We examined the frequency of multiple vertebral fractures (MVFs) and esophageal hiatal hernia (HH) in 18 Japanese postmenopausal women (74.1 ± 9.9 years, mean ± SD), with refractory reflux esophagitis (RRE) that had needed a proton pump inhibitor for more than 6 months to suppress symptoms such as heartburn and acid regurgitation, as well as in 57 control subjects without RE (71.4 ± 5.9 years). MVFs (two or more VFs), HH, and both features were found in 11 (61%), 16 (89%), and 11 (61%) subjects, respectively, in the RRE group. All 11 patients with MVFs also had HH, suggesting their strong association. On the other hand, MVFs, HH, and both were found in 15 (26%), 23 (40%), and 8 (14%) subjects, respectively, in those without RE. The differences in frequencies of MVFs, HH, and both between the two groups were significant (χ2 = 7.3, 12.9, and 16.0; P = 0.015, 0.0009, and 0.0002, respectively). When univariate logistic regression analysis was performed with the presence of RRE as a dependent variable and the presence of MVFs, HH, and both as independent variables, MVFs, HH, and both were selected as indices affecting the presence of RRE (age-adjusted odds ratios: 4.34, 11.07, and 10.30; 95% confidential intervals: 1.40–13.45, 2.30–53.22, and 2.96–35.86; P = 0.0109, 0.0027, and 0.0002, respectively). These results show that the presence of MVFs is associated with the presence of RRE in Japanese postmenopausal women, and this association becomes more significant when HH is present. Thus, a kyphotic lumbar spine with MVFs may cause HH and RE by raising the intraabdominal pressure. As recent therapeutic agents for osteoporosis, alendronate and risedronate, are known to be very effective for suppressing the occurrence of new VFs, these agents may also prevent gastrointestinal disorders such as HH and RRE in osteoporotic women when administered to subjects without VFs.

Quantitative computed tomography of lumbar vertebrae in Japanese patients with osteoporosis

Vertebral trabecular bone mineral density of both healthy Japanese subjects and Japanese patients with osteoporosis was measured by quantitative computed tomography (QCT) technique. The age-related reduction rate in vertebral trabecular bone mineral density of control females averaged 1.1% per year, from age 20 to 80, with an accelerated loss demonstrated after age 40. In the male controls, trabecular bone mineral density declined by an average of 0.9% per year. These values were found to be similar to the University of California at San Francisco (UCSF) QCT data (US Caucasians: 1.2% for female, 0.72% for male). Therefore, it appears that age-related rates of trabecular bone loss in the lumbar vertebrae may be similar for both Japanese and Caucasians. However, when compared to average values in UCSF QCT data of comparable age and sex, the mean values in Japanese appear to be approximately 10-20 mg/cm3 lower than Caucasian counterparts. All female patients with osteoporosis as evidenced by atraumatic vertebral fracture had QCT values below 50 mg/cm3. It is suggested that the Japanese may have lower trabecular bone mineral density than Caucasians but may also have a lower threshold for fracture of the vertebrae. Further studies are needed to establish the possible racial differences in vertebral trabecular bone mineral density, and to determine whether these possible disparities are related to genetic differences, or to differences in body size, dietary intake, physical activity or other lifestyle/environmental factors.

Effects of human PTH-related peptide and human PTH on cyclic AMP production and cytosolic free calcium in an osteoblastic cell clone

Recently, human parathyroid hormone-related peptide (hPTHrP) has been purified and its amino acid sequence determined. Within the amino-terminal 13 residues of hPTHrP, 8 amino acids were found homologous between hPTHrP and human PTH (hPTH). This peptide was reported to stimulate cyclic AMP (cAMP) production in osteoblastic cell lines (UMR106 and ROS17/2.8). However, whether or not this peptide affects another second messenger, i.e., cytosolic free calcium ([Ca2+]i), in osteoblasts has not yet been determined. Therefore, in the present study, we examined the effects of synthetic amino-terminal fragments of hPTHrP (Tyr40hPTHrP1-40 and hPTHrP1-34) on intracellular cAMP production and [Ca2+]i in an osteoblastic cell line (MC3T3-E1) and compared them with those of hPTH1-34. Human PTHrP1-34, Tyr40hPTHrP1-40 and hPTH1-34 stimulated cAMP production in an equipotent manner at concentrations ranging from 2.5 x 10(-10) to 1.3 x 10(-6) M. Human PTH1-34 at concentrations from 2.5 x 10(-7) to 1.3 x 10(-6) M significantly (P less than 0.05) increased [Ca2+]i, but hPTHrP1-34 and Tyr40hPTHrP1-40 at the same concentrations did not. These results suggest a different receptor-mediated mechanism for [Ca2+]i increase between hPTHrP and hPTH, although these two peptides appear to share the same receptor site(s) which is coupled to the cAMP system in MC3T3-E1.

MENOPAUSE-RELATED CHANGES IN BONE MINERAL DENSITY IN JAPANESE WOMEN : A LONGITUDINAL STUDY ON LUMBAR SPINE AND PROXIMAL FEMUR

We investigated 2-year longitudinal changes of bone mineral density (BMD) in lumbar spine and proximal femur in 64 Japanese women aged 38–67. Forty subjects were premenopausal (mean age 44.9) and 24 postmenopausal (mean age 54.6) at enrollment of the study. Six subjects experienced menopause during the 2-year study period and were defined as the perimenopausal group. Measurements of BMD were performed using dual-energy X-ray absorptiometry at L2–4, femoral neck, greater trochanter, and Wards triangle. Paired t test revealed no significant decrease in BMD at any site in the premenopausal group. Significant annual decrease in BMD was observed in the perimenopausal group at L2–4, femoral neck, and greater trochanter. A similar tendency was observed in Wards triangle, but did not reach statistical significance. In the postmenopausal group, significant decrease in BMD was found at the proximal femur, but not at L2–4. Significant inverse correlation between age and change rate of BMD was found at L2–4, but not at the proximal femur, in premenopausal women. In postmenopausal women, there was a significant association between body weight (BW) change and change rate in BMD at L2–4, femoral neck, or greater trochanter. This association was not found in the premenopausal group. These results suggest that effect of menopause on BMD may be different in individuals and sites of the skeleton. BW change may affect change in BMD in postmenopausal women. However, the limited variability in both BW and BMD changes among premenopausal women in this study may explain the poor association between change in BW and change in BMD in the premenopausal group. As individual differences in each group is considerably large, annual measurements of BMD may be necessary to find possible candidates for early intervention.

Femoral and spinal bone mineral density in Japanese osteoporotics with hip fracture

In the present study, bone mineral density (BMD) of femoral neck and lumbar spine was compared between 38 Japanese female patients with hip fracture (age 63–89 years, mean±SD 76±7 years) and 162 age-matched female controls (age 62–90 years, mean±SD 75±7 years). BMD was measured in the femoral neck and lumbar spine (L2–4) using dual-photon absorptiometry (Norland model 2600). BMD values of femoral neck as well as lumbar spine were significantly lower in patients with hip fracture than in controls (0.504±0.097 v 0.597±0.101,p<0.01, for femoral neck; 0.661±0.146 v 0.720±0.128,p<0.05, for lumbar spine). Patients with hip fracture and controls were stratified according to their BMD levels at two measuring sites, and the ratio of the number of patients and controls at each BMD level was calculated as an indicator of fracture rate. This ratio showed an exponential increase as the femoral neck BMD declined, but only a gradual increase as the lumbar spine BMD declined. Specificity-sensitivity analysis revealed that BMD values of 0.59 and 0.54 g/cm2 at the femoral neck provided a specificity of 52% and 68% with a sensitivity of 90% and 75%, respectively. These findings suggest that Japanese patients with hip fracture are more osteoporotic than age-matched controls and that the selective measurement of femoral neck would be useful for predicting the risk of hip fracture.

Quantitative computed tomography: Comparison of two calibration phantoms

Vertebral trabecular mineral density of healthy Japanese (91 females and 67 males) was measured using the quantitative computed tomography (QCT) technique in a cross-sectional study. We compared estimates of vertebral bone density using two calibration phantoms: the Genant K2HPO4 phantom developed by Genant and a solid CaCO3 phantom developed by Chugai Pharmaceutical. Using the CaCO3 phantom, the rate of decrease of spinal trabecular mineral with age in control females averaged 1.1% per year from age 20 to 80, with an accelerated loss demonstrated at menopause (1.8%). Trabecular bone mass in male controls declined an average 0.9% per year. There was a highly significant correlation between the results obtained with each phantom (r=0.980, p<0.001). This relation was linear over the range of bone mass (60 –172 mg K2HPO4/cm3), and was expressed by the equation y(CaCO3)=1.26×(K2HPO4)+12.3 There was, however, some dispersion of the data around the regression. The calibration phantom used for the measurement of the vertebral trabecular bone should need to be more consistent in longitudinal studies. Apart from this consideration, a solid Chugai phantom can be said to be useful having practical advantages in its flexibility.

Radial bone mineral content of normal Japanese infants and prepubertal children: influence of age, sex and body size

The present study was performed to measure appendicular bone mass of Japanese infants and children, and to assess the influence of age, sex and body size on bone mass during the period of bone growth. The bone mineral content (BMC) and bone width (BW) at the distal third of the radius were measured by single photon absorptiometry (SPA) in 229 healthy Japanese infants and children aged 0-12 years, and the BMC/BW ratio was calculated to give the bone mineral density (BMD). BMC and BW increased with age until 2 years, while BMD did not obviously change until 2 years. After 2 years of age, the overall effect of aging appeared more prominent in BMC and BMD than in BW. There were no significant differences in BMC, BW and BMD between males and females aged 0-12 years. Age, body height, and body weight were strongly correlated with three parameters of bone mass (BMC, BW, and BMD). Among the three parameters of bone mass, BMC showed the highest Pearson coefficient of correlation with age (r = 0.955), body height (r = 0.957) and body weight (r = 0.966), as compared with BW and BMD. The present cross-sectional study provides normative data of the appendicular bone mass in healthy Japanese children, which may serve as a standard for assessment of bone mineralization in Japanese infants and children with medical problems.

Basic and clinical evaluation of the measurement of bone resonant frequency

SummaryA new computerized apparatus was constructed to measure the resonant frequency of human ulnain vivo with high sensitivity and reproducibility. Experimental studies using aluminum bar and dried human bone revealed the importance of the ulna being positioned parallel to the radius, approximately 90 degree flexion of the elbow joint, and minimal muscle activity in order to demonstrate maximum resonant frequency of ulna. Measurement of bone resonance in monkeysin vivo and after removal of the bonein vitro showed good agreement. Product of F (maximum resonant frequency in Hz) and L (ulnar length in cm), FL, indicating the speed of propagation of sound wave through the ulna, showed a significant positive correlation with bone mineral content/bone width (BM/BW) measured by Norland-Cameron apparatus and age-bound decline in both sexes. Patients with osteomalacia and primary hyperparathyroid bone disease tended to have higher FL values than expected from BM/BW. Two-dimensional display of FL and BM/BW thus appears to be useful in distinguishing osteoporosis from osteomalacia better than the use of BM/BW alone.

Arachidonic acid stimulates cell growth in an osteoblastic cell line, MC3T3-E1, by noneicosanoid mechanism

SummaryArachidonic acid, added to α-minimum essential medium containing 10% fetal bovine serum at the final concentration of 10−4 M, significantly increased DNA content of an osteoblastic cell line, MC3T3-E1, along with an increase of DNA synthesis. No growth-stimulatory effect of arachidonic acid was observed under serum-free condition. α-Linolenic acid, which cannot be converted to arachidonic acid, also increased DNA content at 10−4 M. Additionally, the stimulatory effects of these fatty acids were not inhibited by simultaneous addition of 10−5 M of indomethacin. Indomethacin, when added to α-minimum essential medium with 10% fetal bovine serum, also significantly increased DNA content of MC3T3-E1 cells. These results suggest that arachidonic acid may potentiate the growth-stimulatory effect of serumderived growth factors probably via noneicosanoid mechanism. Rat osteogenic sarcoma cell line, UMR106, also showed an increase in DNA content with arachidonic acid treatment. Hence, it is suggested that arachidonic acid may stimulate proliferation of cells of osteoblastic lineage. It is also suggested that indomethacin, probably by blocking endogenous prostaglandin E2 synthesis, stimulates cell growth in MC3T3-E1 cells.

Possible involvement of inositol phosphates and calmodulin in calcitonin-induced stimulation of phosphate transport in LLC-PK1 cells

The present study investigated the possible involvement of phosphatidylinositol breakdown and Ca2+-calmodulin complex in the calcitonin-induced stimulation of phosphate transport in LLC-PK1 cells. This cell line with calcitonin receptors possesses Na+-dependent phosphate transport and has been employed as a model for studying the mechanism of renal tubular phosphate transport. (Asu1,7) eel calcitonin stimulated the phosphate transport in LLC-PK1 cells in a dose-dependent fashion with accompanying increase of inositol triphosphate (IP3) production. When the cells were preincubated with the potent calmodulin antagonist, w-7 or w-13, the stimulatory effect of calcitonin on phosphate transport was significantly inhibited. These findings indicate that Ca2+-calmodulin complex formed by increased cytosolic Ca2+, which is mobilized from intracellular pools by IP3, may be involved in the signal transduction of calcitonin in these cells.