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Biomedical subjects

I Owan

Publications and source records attributed to I Owan.

At least 19 recordsLinked to original sources
PubMed1998

Recruitment and proliferative responses of osteoblasts after mechanical loading in vivo determined using sustained-release bromodeoxyuridine.

Mechanical bending of a rat's tibia in vivo can increase endocortical bone formation by over sixfold. It has been proposed that mechanical loading increases bone formation by driving osteoprogenitor cells in the marrow stroma to progress through the cell cycle and subsequently differentiate into osteoblasts at the cortical bone surfaces. We used a sustained-release preparation of 5-bromo-2'-deoxyuridine (SR-BrdUrd) to determine the origin of endocortical osteoblasts in rat tibiae after mechanical loading. SR-BrdUrd was bioavailable for the entire 96 h duration of the experiments, so all cells that progressed through a cell cycle were labeled with BrdUrd. Although the endocortical osteoblast surface was significantly increased (p < 0.05) at 48 h after loading, the percentage of BrdUrd-labeled osteoblasts did not increase, suggesting that the newly differentiated osteoblasts on the endocortical surface did not originate from proliferating cells. At 96 h after loading, 30-40% of the endocortical osteoblasts were BrdUrd labeled. The majority of BrdUrd-labeled osteoblasts appeared on the endocortical bone surface within the third day after loading, indicating that proliferation and differentiation of precursors into endocortical osteoblasts required 72 h after the loading stimulus. These results indicate that mechanical loading can cause two distinct osteoblastic responses: an immediate response within 48 h in which osteoblasts are recruited from nondividing preosteoblasts and/or bone-lining cells, and a delayed response involving proliferation and differentiation of preosteoblasts that requires > or =3 days.

Animals↗
PubMed1997

Effects of nitric oxide synthase inhibitors on bone formation in rats.

NO is synthesized from L-arginine by at least three isoforms of nitric oxide synthase (NOS) and is known to function as a vasodilator and neurotransmitter. NO is produced by bone cells but its function in bone biology is, as yet, unclear. We hypothesized that NOS mediates bone formation in remodeling regions of the skeleton. We studied the effects of two NOS inhibitors: N(G)-nitro-L-arginine methyl ester (L-NAME), which is a general inhibitor of NOS activity and is known to inhibit the vasodilatory effects of the endothelial NOS (eNOS) isoform; and aminoguanidine, which is a selective inhibitor of the inducible NOS (iNOS) isoform. Our hypothesis was tested by treating rats with NOS inhibitors and measuring bone formation rates in the tibial epiphysis and diaphysis. Bone formation indices were measured using standard bone histomorphometry. L-NAME treatment significantly raised mean arterial blood pressure (MAP). This effect was partially reversed by addition of L-arginine. Aminoguanidine had no significant effect on MAP, indicating that it did not block eNOS. The treatments also had substantial effects on bone formation in remodeling trabecular bone. L-NAME did not significantly change trabecular bone formation rate, whereas aminoguanidine reduced bone formation rate in the tibial epiphysis by 79% compared with control. This reduction was completely reversed by L-arginine, suggesting that bone formation during remodeling is, in part, mediated through L-arginine metabolism. No effect of aminoguanidine on bone formation was seen in the tibial diaphysis, a site that undergoes minimal bone remodeling. This finding suggests that the L-arginine-NO pathway is important in bone remodeling.

Animals↗
PubMed1997

Mechanotransduction in bone: osteoblasts are more responsive to fluid forces than mechanical strain.

Mechanical force applied to bone produces two localized mechanical signals on the cell: deformation of the extracellular matrix (substrate strain) and extracellular fluid flow. To study the effects of these stimuli on osteoblasts, MC3T3-E1 cells were grown on type I collagen-coated plastic plates and subjected to four-point bending. This technique produces uniform levels of physiological strain and fluid forces on the cells. Each of these parameters can be varied independently. Osteopontin (OPN) mRNA expression was used to assess the anabolic response of MC3T3-E1 cells. When fluid forces were low, neither strain magnitude nor strain rate was correlated with OPN expression. However, higher-magnitude fluid forces significantly increased OPN message levels independently of the strain magnitude or rate. These data indicate that fluid forces, and not mechanical stretch, influence OPN expression in osteoblasts and suggest that fluid forces induced by extracellular fluid flow within the bone matrix may play an important role in bone formation in response to mechanical loading.

3T3 Cells↗
PubMed1996

High fluoride intakes cause osteomalacia and diminished bone strength in rats with renal deficiency.

Renal insufficiency is known to increase plasma fluoride levels, which may increase the risk of fluorosis and osteomalacia. The purpose of this study was to determine the effects of fluoride on skeletal fragility and mineralization in renal-deficient animals. We evaluated the skeleton of rats with surgically induced renal deficiency (4/5 nephrectomy) that were chronically exposed to fluoridated water at concentrations of 0, 5, 15, and 50 ppm for a period of 6 months. The chosen fluoride doses caused plasma fluoride levels equivalent to those in humans consuming fluoridated water levels of 0, 1, 3, and 10 ppm, respectively. Animals with renal deficiency drank about 60% more water and excreted 85% more urine than control animals. Glomerular filtration rate (GFR) was decreased 68% and plasma BUN was increased fourfold in rats with renal deficiency. Plasma fluoride was strongly correlated with 1/GFR and was greatly increased by renal deficiency in all animals consuming fluoridated water. There was a strong positive, nonlinear relationship between plasma fluoride and bone fluoride levels, suggesting nonlinear binding characteristics of fluoride to bone. The amount of unmineralized osteoid in the vertebral bone was related to the plasma fluoride levels. Vertebral osteoid volume was increased over 20-fold in animals with renal deficiency that received 15 or 50 ppm fluoride, suggesting osteomalacia. Should osteomalacia be defined as a tenfold increase in osteoid volume, there appeared to be a threshold plasma fluoride level of about 20 micromol/L, above which osteomalacia was observed consistently. This plasma fluoride level was not achieved in control rats regardless of fluoride intake, nor was it achieved in renal-deficient rats receiving 0 or 5 ppm fluoride. A fluoride concentration of 50 ppm reduced femoral bone strength by 11% in control rats and by 31% in renal-deficient rats. Vertebral strength also was decreased significantly in renal-deficient rats given 50 ppm fluoride. In conclusion, fluoridated water in concentrations equivalent to 3 and 10 ppm in humans, caused osteomalacia and reduced bone strength in rats with surgically-induced renal deficiency.

Alkaline Phosphatase↗
PubMed1996

Increased bone formation in rat tibiae after a single short period of dynamic loading in vivo.

Based on our quantum concept for mechanically adaptive bone formation, we hypothesized that a single bout of loading would increase bone formation at the endosteal surface in rat tibiae, with a maximal response 4-8 days after loading and a stimulus-response relationship for load magnitude. Bending loads were applied to right tibiae of rats at 31, 43, 53, or 65 N for a single bout of 36 or 360 cycles; bone formation was assessed 1-4, 5-8, or 9-12 days after loading. A single loading episode increased lamellar bone formation rate (BFR) in all groups (P<0.05) and was maximal 5-8 days after loading. A distinct dose-response relationship was not evident among all load magnitudes or for duration, but 65 N was significantly more osteogenic than loads of 31-53 N (P<0.05), consistent with a threshold response to loading. There was also evidence for a significant increase in BFR (P<0.05) and double-labeled surface (P<0.01) within 4 days of loading, suggesting that bone-lining cells were activated directly by the stimulus. Thus subtle changes in BFR may occur by modulating the activity of surface cells, but large modeling drifts and anabolic responses require recruitment and differentiation of osteoprogenitor cells near the bone surface.

Analysis of Variance↗
PubMed1996

Nitric oxide inhibitor L-NAME suppresses mechanically induced bone formation in rats.

We hypothesized that nitric oxide may act as an intermediary in the transduction of mechanical loading of bone into a bone formation response. In the present study, 48 rats were divided into the following three treatment groups: control, treatment with N omega-nitro-L-arginine methyl ester (L-NAME; an inhibitor of nitric oxide synthase), and treatment with D-NAME (the less active enantiomer of L-NAME). The rats were subdivided into groups subjected to four-point bending or sham loading of their right tibiae. Bone formation was measured at the midshaft of the loaded and nonloaded (left) tibiae of each rat using histomorphometric methods. The application of four-point bending, but not sham loading, resulted in new bone formation. Treatment with L-NAME reduced the rate of mechanically induced bone formation by 66% compared with the control group. Bone formation rates in nonloaded or sham-loaded limbs were not affected by L-NAME treatment. The results suggest that nitric oxide may play a role in the transduction of a mechanical stimulus into a biological response in bone.

Animals↗
PubMed1996

Advantages of raloxifene over alendronate or estrogen on nonreproductive and reproductive tissues in the long-term dosing of ovariectomized rats.

For the first time, raloxifene or alendronate was administered to rats immediately after ovariectomy for 10 months and compared with estrogen to elucidate mechanisms behind the raloxifene effects observed in nonreproductive and reproductive tissues. Specifically, 75-day-old rats were randomly selected as sham controls (Sham), ovariectomized controls (Ovx) or ovariectomized rats treated with fully efficacious doses of raloxifene (RA), 17 alpha-ethynyl estradiol (EE2) or alendronate (ABP). Lumbar vertebrae and proximal tibiae were examined by computed tomography (QCT) and by histomorphometry. Histomorphometry showed differences in bone architecture between groups when QCT densities were similar, but tibial trabecular bone analysis by QCT correlated with histomorphometry with r = .86 to .93, depending on the parameter. Both techniques confirmed that Ovx had substantially less bone than Sham, with greater loss of trabecular bone in the proximal tibia than vertebrae. Both techniques showed that RA had effects similar to but not identical with EE2 in preventing bone loss in vertebrae and tibiae. ABP partially prevented loss of bone in L-5, but was not significantly different from Ovx in the proximal tibia. This may be caused by ABP suppression of bone apposition, beyond effects observed for EE2 or RA. RA appeared to be more similar to EE2 because ABP significantly depressed bone formation (bone formation rate, mineral apposition rate) to below RA or EE2 levels, especially in L-5. Mechanical loading to failure of L-6 vertebrae showed a rank order of vertebral strength of Sham > RA > EE2 > Ovx > ABP, although significant differences were not observed between treatment groups. These data show that ABP suppression of bone formation can affect bone quality with long-term treatment. In other tissues, RA had minimal uterine effects, while significantly lowering serum cholesterol to below EE2-treated levels. Both EE2 and RA rats had significantly lower body weights than the other groups. ABP had no effect on serum lipids, uterine weight or body weight. Therefore, RA appears to have a broader range of desirable effects on bone, body weight, uteri and cholesterol than ABP or EE2 in ovariectomized rats.

Alendronate↗
PubMed1995

Aging changes mechanical loading thresholds for bone formation in rats.

The effect of aging on the mechanical loading thresholds for osteogenesis was investigated in rats. We applied mechanical loads varying from 30 to 64 N to the tibiae of 43 19-month-old rats using a four-point bending apparatus. Bone formation rates were measured on the periosteal and endocortical surfaces of the tibial midshaft using double-label histomorphometry. Bone formation rates from the old rats were compared with results from adult (9-month-old) rats that we reported earlier.(4) Bone formation on the periosteal surface of the old rats was predominantly woven-fibered. Periosteal bone formation was observed in a lower percentage of the old rats compared with the younger adult rats for applied loads of 40 N and greater (59% old, 100% adult). However, in the old rats that formed woven bone there were no significant differences in woven bone area (p=0.1) or surface (p=0.24) compared with younger adults. Therefore, the periosteum of old rats had a higher threshold for activation by mechanical loading, but after activation occurred, the cells had the same capacity to form woven bone as younger adult animals. On the endocortical surface, relative bone formation rates in old rats showed a marginal (p=0.06) increase in response to an applied load of 64 N but was not increased at lower loads. The relative bone formation rate in the old rats was over 16-fold less than that reported for the younger adult rats at an applied load of 64 N and the relative bone forming surface in old rats in this study was 5-fold less than it was younger rats under similar loading conditions. In the younger adult rats, a mechanical threshold for lamellar bone formation of 1050 microstrain was calculated for the endocortical bone surface. The old rats required over 1700 microstrain on the endocortical surface before bone formation was increased. The data suggest that both the periosteal and endocortical surfaces of the tibiae of older rats are less responsive to mechanical stimuli.

Adaptation, Physiological↗
PubMed1995

Mechanotransduction in bone: role of strain rate.

Bone tissue can detect and respond to its mechanical environment, but there is no consensus for how bone cells detect mechanical loads. Some think that cells sense tissue deformation (strain) and respond when strain is abnormally high. However, strains in bone tissue are usually very small, and it is questionable whether bone cells are sensitive enough to detect them. Another theory suggests that mechanical loads are coupled to the bone cells by stress-generated fluid flow within the bone tissue, which is dependent on the rate of change of bone strain. We applied bending loads to the tibiae of adult rats to create equivalent peak strains in the bone tissue but with varied rates of strain. Bone formation was significantly increased in the two experimental groups when the highest strain rates were compared with lower strain rates (P < 0.01), and the amount of new bone formation was directly proportional to the rate of strain in the bone tissue. These results suggest that relatively large strains alone are not sufficient to activate bone cells. High strain rates and possibly stress-generated fluid flow are required to stimulate new bone formation.

Adaptation, Physiological↗
PubMed1995

Type 1-like helper T cell lines responsive to autologous peripheral blood monocytes established from two patients with sarcoidosis.

In the present study, T cell lines, designated TU/BAL and KC/LN, were established from bronchoalveolar lavage (BAL) fluid and an affected lymph node, respectively, obtained from patients with active sarcoidosis by cultivating in the presence of IL-2. These cell lines produced IL-2 and proliferated by stimulation with the patient's own peripheral blood mononuclear cells treated with mitomycin C, while three other T cell lines established from ConA-stimulated BAL cells of patients with non-sarcoid lung diseases did not show any proliferative responses. The proliferation was mediated by IL-2, because anti-IL-2R alpha-chain monoclonal antibody (mAb) inhibited this response in a dose-dependent fashion. The adherent cell was a main stimulator of the proliferation. Both CD4 and HLA-DR appeared to be involved, because mAbs against these molecules inhibited this response. These results suggest that T cells obtained from sarcoid patients respond to a certain unknown antigen associated with HLA-DR or some self antigen expressed on the monocytes. Furthermore, both TU/BAL and KN/LN represented a profile of Th1-like cells: they secreted IL-2 and IFN-gamma, but not IL-4, IL-5 and IL-6, when stimulated with PHA in the presence or absence of 12-O-tetradecanoylphorbol-13-acetate. Thus, Th1-like cells activated by some unknown antigen(s) might play roles in the pathogenesis of sarcoidosis.

Adult↗
PubMed1994

Postmitotic osteoclast precursors are mononuclear cells which express macrophage-associated phenotypes.

We previously demonstrated that osteoclast-like multinucleated cells were formed within 6 days in cocultures of mouse osteoblastic cells and spleen cells in response to 1 alpha,25-dihydroxyvitamin D3[1 alpha,25(OH)2D3] which was added together with hydroxyurea on Days 4-6 (final 2 days of the 6-day coculture period). Using this coculture system, chronological changes of macrophage-associated phenotypes such as nonspecific esterase (NSE) and antigens to Mac-1, Mac-2, and F4/80 were examined in postmitotic osteoclast precursors during differentiation into osteoclasts induced by 1 alpha,25(OH)2D3 (10 nM) added on Day 4. Osteoclast differentiation was assessed by examining expression of calcitonin receptors (CTRs) by autoradiography using 125I-labeled salmon CT. CTRs were first detected on small mononuclear cells within 12 hr after adding 1 alpha,25(OH)2D3. The number of CTR-positive mononuclear cells attained a maximum at 24 hr and decreased thereafter. CTR-positive multinucleated cells were first observed at 24 hr and reached a maximum population at 48 hr. All CTR-positive cells showed tartrate-resistant acid phosphatase activity (a marker enzyme of osteoclasts). Most of the CTR-positive mononuclear cells which appeared at 12 hr were positive for NSE and antigens to Mac-1 and Mac-2, but negative for F4/80 antigen. The proportion of CTR-positive cells expressing NSE and Mac-1 to total CTR-positive mononuclear cells decreased time dependently. Like authentic osteoclasts, CTR-positive multinucleated cells were negative for NSE and antigens to Mac-1 and F4/80, but positive for Mac-2. These results indicate that postmitotic osteoclast precursors are mononuclear phagocytes with macrophage-associated phenotypes, some of which disappear rapidly during their differentiation into osteoclasts.

Acid Phosphatase↗
PubMed1994

The role of calcitonin gene-related peptide (CGRP) in macrophages: the presence of functional receptors and effects on proliferation and differentiation into osteoclast-like cells.

It has been shown that both calcitonin gene-related peptide (CGRP) and amylin bind weakly to calcitonin (CT) receptors in osteoclast-like cells formed in vitro and inhibit bone resorption by a cAMP-dependent mechanism. Osteoclasts are thought to be derived from cells of the monocyte macrophage lineage, in which CGRP, but not CT, induces cAMP production. In this study, we determined the presence of functional receptors for CGRP in mouse alveolar macrophages and the effects of this peptide on proliferation and osteoclastic differentiation in mouse alveolar and bone marrow-derived macrophages. Human CT did not stimulate cAMP production in macrophages. Human CGRP stimulated cAMP production in mouse alveolar macrophages and bone marrow-derived macrophages dose-dependently. Human amylin, which has 43% homology with human CGRP, also stimulated these macrophages to produce cAMP, but only at a 100-fold higher concentration. The increment in cAMP production induced by human CGRP and amylin was abolished by the addition of human CGRP(8-37), a selective antagonist for CGRP receptors. Specific binding of [125I]human CGRP to alveolar macrophages was detected (dissociation constant, 2.5 x 10(-8) M; binding sites, 1.4 x 10(4)/cell). Amylin, but not CT, displaced the bound [125I]human CGRP from alveolar macrophages, but at a 100-fold higher concentration. No specific binding of [125I]human CT and [125I]human amylin to alveolar macrophages could be detected. Pretreatment with human CGRP for 24 h dose-dependently suppressed DNA synthesis in alveolar macrophages induced by granulocyte-macrophage colony-stimulating factor (GM-CSF). CGRP also suppressed the number of macrophage colonies formed from bone marrow cells induced by macrophage colony-stimulating factor (M-CSF).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

[Serodiagnosis of Legionella pneumonia--data of our laboratory in the recent 3 years].

We had examined antibody titers against Legionella spp. of patients' sera which were mainly sent from other hospitals, performed with the indirect fluorescent antibody (IFA) method. The clinical status of the cases diagnosed as Legionella pneumonia serologically, were also studied. Out of 105 cases with clinically suspected Legionella pneumonia, 15 cases (14.3%) were seropositive. In 9 out of the 15 cases (60.0%) were caused by Legionella pneumophila serogroup 1. Clinical outline of these 15 cases did not contradict those reported in the literature, and erythromycin was effective in many cases. Significant rises (more than four times) of the titer were observed 3 to 4 weeks after onset in most of cases. We would like to emphasize that this should be performed serodiagnosis of Legionella pneumonia.

Adult↗
PubMed1994

[Pulmonary nocardiosis concurrent with Pneumocystis carinii pneumonia in a case of smoldering adult T-cell leukemia].

A 76-year-old man, with concomitant pulmonary nocardiosis and Pneumocystis carinii (PC) pneumonia, is described. He suffered from smoldering type adult T-cell leukemia and strongyloidiasis. Chest X-rays showed bilateral infiltrative shadows. Nocardia asteroides was isolated from sputum and bronchofiberscopic aspirates. PC was not detected, however, despite the chest X-ray raising the suspicion of PC pneumonia. Since PC antigen was detected by immunoelectrophoresis in the absence of other clinical or histologic data, we diagnosed a latent PC infection. He improved with the use of cefuzonam and ST compounds, but experienced recurrence five months after discharge. N. asteroides and PC were detected from sputum on the second admission. As multiple concurrent infections with opportunistic organisms are not uncommon in immunocompromised hosts, we must maintain a high index of suspicion for opportunistic infection in cases such as this one.

Aged↗
PubMed1992

Mechanism of action of amylin in bone.

Amylin is a 37 amino acid peptide produced mainly by beta-cells of the endocrine pancreas. Human amylin has 43% homology with human calcitonin gene-related peptide (CGRP) and 13% homology with human calcitonin (CT). Amylin and CGRP have been reported to have CT-like hypocalcemic activity in vivo. To investigate the role of amylin in bone, we examined the mechanisms of action of human amylin, CGRP, and CT in osteoclasts and osteoblasts. Both human amylin and CGRP inhibited 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3]- induced bone resorption in an organ culture system, and the potencies of the two peptides were similarly approximately 60-fold lower than that of human CT. Using a recently developed procedure for preparing large numbers of osteoclast-like multinucleated cells (MNCs) formed in co-cultures of mouse osteoblasts and bone marrow cells in the presence of 1 alpha,25(OH)2D3, we found that both human amylin and CGRP stimulated cAMP production in osteoclast-like MNCs, but only at 60-fold higher concentrations than human CT. Specific binding of [125I]-human CT to osteoclast-like MNCs was detected (dissociation constant, 3 x 10(-8) M; binding sites, 3 x 10(7) per cell). To displace the bound [125I]-human CT from osteoclast-like MNCs, about 170-fold higher concentrations of human amylin and CGRP were required. No specific bindings of [125I]-amylin and [125I]-CGRP to osteoclast-like MNCs could be detected. Human CGRP stimulated cAMP production both in established mouse osteoblast-like cells (KS-4) and in mouse primary osteoblast-like cells. Amylin was a weak agonist for cAMP production in KS-4 cells. The increment in cAMP production induced by CGRP and amylin was abolished by the addition of human CGRP(8-37), a selective antagonist for CGRP receptors. CT did not stimulate cAMP production in KS-4 cells. Amylin, but not CT, displaced the bound [125I]-human CGRP from rat brain membranes. These results indicate that amylin binds not only to CT receptors in osteoclast-like MNCs but also to CGRP receptors in osteoblasts. The relative potencies of these compounds to induce cAMP production was CT greater than amylin not equal to CGRP in osteoclast-like MNCs and CGRP greater amylin much greater than CT in osteoblast-like cells.

Amyloid↗
PubMed1992

[Detection of Legionella pneumophila using a nested polymerase chain reaction].

We evaluated the usefulness of a Nested PCR method for detecting Legionella pneumophila. This method resulted in L. pneumophila specific detection as far as we evaluated. The first and second step PCR achieved the sensitivity as small as 10 pg and 10 fg of the target DNA, respectively. In the detection from Legionella seeded sputa, the method could detect 0.1 cfu/ml of the bacteria, and it took about 12 hours to detect the target DNA. We demonstrated that the Nested PCR method was superior in sensitivity and rapidity for isolation of the bacteria to the conventional using low pH treatment and selective media for Legionella.

Base Sequence↗
PubMed1992

[Mechanism of acquired resistance against Legionella--blastogenic responses of murine spleen lymphoid cells following the stimulation with Legionella antigens].

We have tried to characterize the blastogenic responses of murine spleen lymphoid cells from BALB/c mice immunized with Legionella pneumophila serogroup (SG) 1 (Philadelphia 1 strain) and non-treated mice. Lymphoid cells from immunized mice showed stronger blastogenic responses following stimulation with concanavalin A or formalin-treated L. pneumophila SG1 whole cell antigen than those showed by lymphoid cells from non-treated mice. These cells from immunized mice also responded strongly when stimulated in vitro with other SGs of L. pneumophila, while these responded weakly when stimulated with other species of Legionella. Serum antibody titers of immunized mice against each SG of L. pneumophila were examined and the cross reactions were also recognized. However, the relatedness of serum antibody titers and the blastogenic responses against each serogroup of L. pneumophila was small. The epitopes recognized by the cellular immunity might be different in part from those recognized by serum antibodies, and investigations should be made on what the cellular immunity recognizes and how it works.

Animals↗