Variation of long-lived free radicals responsible for the EPR native signal in bone of aged or diseased human females and ovariectomized adult rats.

The purpose of this study was to gain insights into the variations seen in the electron paramagnetic resonance (EPR) spectroscopy of the native signals of teeth and bones used for retrospective dosimetry measurements. We determined that changes occur in the long-lived free radicals responsible for the native signal of cortical bone in aging or diseased human females and aged ovariectomized rats. This was done by measuring the magnitude of the broad (BC) and narrow (NC) components of the native EPR signal of bone following chemical extraction, aging, crushing and thermal annealing. Bone from the upper midshaft of femora of young (17-34 years old, n=5) and elderly (70-92 years old, n=18) females was examined. The results showed that the elderly women had significantly higher BC than the younger women (P<0.01). A similar interpretation was made of the data from an aging female rat osteoporosis model. The results for the NC signals were similar. Finally, dramatic decreases in both NC and BC signals were seen in HIV positive and uncontrolled diabetic (one each) patients indicating the need for studying this signal for a broad spectrum of metabolic disorders. Experiments were performed which strongly indicate that iron liganded with organic molecules is the source of the BC signal. Finally, the accuracy achieved in this study indicates that resolving the dosimetric signal (g=2.0018) should be improved by subtraction of the deconvoluted NC and BC signals from the original spectrum.

Determining mineral content variations in bone using backscattered electron imaging.

The mechanical properties of bones are greatly influenced by the ratio of organic constituents to mineral. Determination of bone mineral content on a macroscopic scale is straightforward, but microscopic variations, which can yield new insights into remodelling activities, mechanical strength, and integrity, are profoundly more difficult to measure. Measurement of microscopic mineral content variations in bone material has traditionally been performed using microradiography. Backscattered electron (BSE) imaging is a technique with significantly better resolution than microradiography with demonstrated consistency, and it does not suffer from projection-effect errors. We report results demonstrating the applicability of quantitative BSE imaging as a tool for measuring microscopic mineral content variations in bones representing a broad range of mineralization. Bones from ten species were analyzed with Fourier-transformed infrared spectroscopy, X-ray diffraction, energy dispersive X-ray spectrometry, ash measurements, and BSE imaging. BSE image intensity (graylevel) had a very strong positive correlation to mineral (ash) content. Compositional and crystallographic variations among bones had negligible influence on backscattered electron graylevels. The present study confirms the use of BSE imaging as a tool to measure the microscopic mineral variability in a broad range of mineralized tissues.

Swim-trained rats have greater bone mass, density, strength, and dynamics.

Weight-bearing exercise is traditionally recommended for improving bone health in postmenopausal women. Effects of swim exercise were studied as an alternative to weight-bearing exercise in ovariectomized rats. Rats in a swim group (Sw, n = 8) swam for 12 wk, 5 days/wk for 60 min per session. A control group (Con, n = 9) engaged in no structured exercise. Femurs were analyzed for bone mineral density and for bone mineral content by dual energy X-ray absorptiometry, biomechanical properties by three-point bending (Instron), and bone structure and formation by histomorphometry. Food intake did not differ among groups. Final body weights were significantly lower in Sw compared with Con (P < 0.05). Swimmers had significantly greater femoral shaft bone mineral density and content (P < 0.05) compared with Con. Femurs of the Sw group had greater mechanical properties (P < 0.05) compared with Con. Histomorphometric data were significantly better in the Sw group compared with Con after the 12-wk intervention (P < 0.05). In conclusion, data from this study demonstrate some beneficial effects of swim exercise on bone structure, turnover, and strength.

Influence of topography and specimen preparation on backscattered electron images of bone.

Backscattered electron (BSE) images of bone exhibit graylevel contrast between adjacent lamellae. Mathematical models suggest that interlamellar contrast in BSE images is an artifact due to topographic irregularities. However, little experimental evidence has been published to support these models, and it is not clear whether submicron topographical features will alter BSE graylevels. The goal of this study was to determine the effects of topography on BSE image mean graylevels and graylevel histogram widths using conventional specimen preparation techniques. White-light interferometry and quantitative BSE imaging were used to investigate the relationship between the BSE signal and specimen roughness. Backscattered electron image graylevel histogram widths correlated highly with surface roughness in rough preparations of homogeneous materials. The relationship between BSE histogram width and surface roughness was specimen dependent. Specimen topography coincided with the lamellar patterns within the bone tissue. Diamond micromilling reduced average surface roughness when compared with manual polishing techniques but did not significantly affect BSE graylevel histogram width. The study suggests that topography is a confounding factor in quantitative BSE analysis of bone. However, there is little quantitative difference between low-to-moderate magnification BSE images of bone specimens prepared by conventional polishing or diamond micromilling.

Errors in quantitative backscattered electron analysis of bone standardized by energy-dispersive x-ray spectrometry.

Backscattered electron (BSE) imaging has proven to be a useful method for analyzing the mineral distribution in microscopic regions of bone. However, an accepted method of standardization has not been developed, limiting the utility of BSE imaging for truly quantitative analysis. Previous work has suggested that BSE images can be standardized by energy-dispersive x-ray spectrometry (EDX). Unfortunately, EDX-standardized BSE images tend to underestimate the mineral content of bone when compared with traditional ash measurements. The goal of this study is to investigate the nature of the deficit between EDX-standardized BSE images and ash measurements. A series of analytical standards, ashed bone specimens, and unembedded bone specimens were investigated to determine the source of the deficit previously reported. The primary source of error was found to be inaccurate ZAF corrections to account for the organic phase of the bone matrix. Conductive coatings, methylmethacrylate embedding media, and minor elemental constituents in bone mineral introduced negligible errors. It is suggested that the errors would remain constant and an empirical correction could be used to account for the deficit. However, extensive preliminary testing of the analysis equipment is essential.

Spaceflight alters bone mechanics and modeling drifts in growing rats.

BACKGROUND: Alterations in bone metabolism may be a particularly serious consequence of spaceflight and a major obstacle to long-term space exploration. The effects of spaceflight on bone mechanics are unclear. This study examined the effects of spaceflight on bone mechanics in a growing rat model during a 17-d mission aboard the space shuttle (STS-78). METHODS: There were 18 rats that were divided into 3 experimental groups: flight rats (n = 6), ground-based control rats housed in an animal enclosure module (AEM, n = 6), and ground-based control rats housed in standard vivarium caging (n = 6). At the conclusion of the mission, rat femurs were tested in three-point bending followed by static and dynamic bone histomorphometry. RESULTS: Maximum stress was unaffected by spaceflight, but flexural rigidity was significantly decreased in flight animals. Much of the decrease appeared to be the result of decreases in tissue properties (elastic modulus) rather than structural changes within the bone. No significant differences in cortical bone mass or geometry were observed. In contrast, endocortical resorption was significantly decreased in flight rats accompanied by a nonsignificant decrease in periosteal bone formation, suggesting alterations in bone modeling drifts during spaceflight. For nearly all measured indices, ground-based AEM rats displayed values intermediate to flight and ground-based vivarium rats. CONCLUSIONS: Spaceflight can impair tissue properties in femoral cortical bone during growth without significant decreases in bone mass or geometry.

[Solitary plasmacytoma of the lung]. / Soliter tüdö plasmacytoma.

A case of solitary plasmacytoma of the lung in a 64 year old woman is reported. The anatomical and clinical features of this exceedingly rare form are discussed. Stress is laid on the 'fact that diagnostic certainty can only be obtained by immunohistochemistry. It is also necessary that patients be subjected to periodical radiological and immunological studies in the postoperative period to detect the possible occurrence of multiple myeloma.

[Pulmonary echinococcosis]. / Pulmonalis echinococcosis.

Two patients with pulmonal Echinococcosis are presented by the authors. Contact with cestoda carrier animals cannot be revealed. At the first patient, who was without any complaints, the two round shadows in her chest X-ray were detected accidentally by screening examination and before the operation only eosinophilia, many eosinophil granulocytes in the pleural effusion and the positive complement binding reaction were related to the Echinococcosis, which was proved by the histological examination of the intraoperative biopsy. The other patient had complaints during months. In this case the Echinococcus granulosus broke into the bronchus and emptied in the sputum. Postoperative histological diagnosis can be obtained by help of the excision from the intraluminal alteration which was visible during bronchoscopy. The authors summarize the etiology, the clinical signs, the diagnose and the therapy of Echinococcosis. They call attention to the incidence of this mainly tropical disease in Hungary and to the rare soliter pulmonal manifestation of the Echinococcosis.

[High-resolution computer tomography of diffuse infiltrative lung diseases]. / Diffúz infiltratív tüdöbetegségek nagy felbontóképességP6 komputertomográfiás vizsgálata.

During a period of 17 months, a total of 101 patients were examined by high resolution computed tomography (HRCT) whose minor focal changes revealed by chest X-ray raised the suspicion of diffuse infiltrative pulmonary disease. The disease of the interstitium was primarily caused by idiopathic pulmonary fibrosis (19%), autoimmune disease (18%), Boeck sarcoidosis (18%) as well as by specific (16%) and aspecific inflammation (12%). HRCT disclosed reticular (61%), nodular (55%), ground-glass opacity-like (48%), emphysematous (33%) and ring-like changes (17%). Also transbronchial excision was performed in 51 patients the results of which were compared to the changes observed during HRCT. Concerning fibrosis and inflammation, HRCT and histological analysis showed identical results in 70%. According to the authors observations, this technique is suitable for detection of fairly fine fibrotic, empysematous and inflammatory signs and for defining the activity of the individual disease as also for the selection of the optimal site of transbronchial excision. HRCT is a useful investigative method both for diagnosing diffuse infiltrative pulmonary disease as well as in following up the effectiveness of treatment.

Age-related hypermineralization in the female proximal human femur.

Hip fracture incidence increases exponentially with age in virtually every human population that has been studied. In spite of this, relatively few studies have examined age-related changes in the metaphyseal cortex of the proximal femur. The present study investigates cortical aging changes in the female proximal femur, with particular reference to regions of hypermineralization. Thirty-three femora from Caucasian females were obtained at autopsy and analyzed using backscattered electron imaging. Variations in hypermineralized tissue area, cortical bone area, and porosity were quantified with standard stereological methods. Cortical width was quantified with digital calipers. Gender differences were examined by statistical comparison with previously published results. Hypermineralized tissue volume was significantly (P < 0.001) greater in elderly individuals. Hypermineralized tissue preferentially appeared near ligamentous or tendinous insertion sites, suggesting the hypermineralized tissue may be a calcified fibrocartilage. Cortical width significantly (P < 0.001) decreased with age and porosity significantly (P < 0.001) increased with age, however the changes were site-specific. The femoral neck and intertrochanteric cortices had a smaller change in cortical width and porosity with age than the diaphysis, but the femoral neck and intertrochanteric cortices had a larger increase in hypermineralized tissue. Comparison with previous data suggests that cortical aging in the proximal femur is similar between males and females and is unlikely to explain the higher incidence of fracture in females. However, the data strongly indicates that age-related changes in the femoral diaphysis cannot be directly extrapolated to either the femoral neck or intertrochanteric cortices.

Evidence of a hypermineralised calcified fibrocartilage on the human femoral neck and lesser trochanter.

Femoral neck fractures are a major cause of morbidity and mortality in elderly humans. In addition to the age-related loss of cancellous bone, changes to the microstructure and morphology of the metaphyseal cortex may be a contributing factor in osteoporotic hip fractures. Recent investigations have identified a hypermineralised tissue on the neck of the femur and trochanteric region that increases in fractional area with advancing age in both males (Boyce & Bloebaum, 1993) and females (Vajda & Bloebaum, 1999). The aim of this study was to determine if the hypermineralised tissue previously observed on the proximal femur is calcified fibrocartilage. Regional variations in the fractional area of hypermineralised tissue, cortical bone, and porosity of the cortical bone along the neck of the femur and lesser trochanter were also quantified. Comparison of back scattered electron and light microscope images of the same area show that regions of hypermineralised tissue correlate with the regions of calcified fibrocartilage from tendon and capsular insertions. The hypermineralised tissue and calcified fibrocartilage had similar morphological features such as the interdigitations of the calcified fibrocartilage into the bone, lacunar spaces, and distinctly shaped pores adjacent to the 2 tissues. Regions of the neck that did not contain insertions were covered with periosteum. There were no regional differences (P > 0.05) on the superior and inferior femoral neck in terms of the percentage area of hypermineralised calcified fibrocartilage, cortical bone, or cortical bone porosity. The lesser trochanter exhibited regional differences in the fractional area of hypermineralised calcified fibrocartilage (P = 0.007) and cortical bone (P = 0.007) but not porosity of the cortical bone (P > 0.05). The effects of calcified fibrocartilage on femoral neck periosteal expansion, repair, and mechanics are unknown, but may play a role in osteoporotic fractures and intracapsular fracture healing.

Cancellous and cortical bone mechanical properties and tissue dynamics during pregnancy, lactation, and postlactation in the rat.

There are substantial changes in maternal skeletal dynamics during pregnancy, lactation, and after lactation. The purpose of this study was to correlate changes in cortical and cancellous bone mass, structure, and dynamics with mechanical properties during and after the first reproductive cycle in rats. Rats were mated and groups were taken at parturition, end of lactation and 8 wk after weaning, and were compared with age-matched, nulliparous controls. Measurements were taken on femoral cortical bone and lumbar vertebral body cancellous bone. At the end of pregnancy, there was an increase in cortical periosteal bone formation and an increase in cortical volume, but a suppression of turnover in cancellous bone with no change in cancellous or cortical mechanical properties. Lactation was associated with a decrease in cortical and cancellous bone strength with a decrease in bone volume, but an increase in turnover on cancellous and endocortical surfaces. After lactation, there was a partial or full restoration of mechanical properties. This study demonstrates substantial changes in bone mechanics that correlate with changes in bone structure and dynamics during the first reproductive cycle in rats. The greatest changes were observed during the lactation period with partial or full recovery in the postlactational period.

Age-related cancellous bone loss in the proximal femur of caucasian females.

The purpose of this investigation was to directly define the age-related intrafemoral variations in cancellous bone density, bone mineralization and rate of bone loss in a cadaveric population of Caucasian female femoral necks and trochanters. Forty-three Caucasian female femora were obtained and divided into premenopausal, postmenopausal and elderly age groups. The neck and trochanter were removed, and cores of cancellous bone were taken from the superior, middle and inferior regions; volume fraction and ash fraction were determined for each core. The cancellous bone volume fraction of the neck was significantly greater than that of the trochanter, as was that of the inferior region of the neck compared with the superior and middle regions at all age groups (p<0.05). The mean neck/trochanter and neck inferior/superior volume fraction ratios did not change with age; however, the variance increased with age (p<0.001). This increasing variability with age suggests that there may be a subpopulation of individuals within the elderly Caucasian population with a significantly different intrafemoral bone density distribution than was present prior to menopause. This study identified no mineralization changes with age in the cancellous bone of the proximal femur (p>0.05). The influence of increased neck/trochanter and neck inferior/superior ratios on femoral neck integrity and fracture prediction is of interest and requires further investigation.

Consistency in calibrated backscattered electron images of calcified tissues and minerals analyzed in multiple imaging sessions.

Pure metal standards have been used to calibrate the operating envionment in quatitative backscattered electron (BSE) imaging of mineralized tissue, allowing comparisons to be made between various mineralization states of bone at the microscopic level. It has not previuously been documented that calibration procedures produce consistent, reliable results over multiple imaging sessions. In this study, BSE images were obtained from bones, pure metals, and a naturally occurring mineral in multiple imaging sessions over a six day period. The graylevel histogram profile (GHP) from each specimen was analyzed for changes in the shape and relative placement on the graylevel spectrum. Computer controlled calibration and a restrospective calibration method using pure aluminum and pure magnesium-aluminum-zinc demonstrated consistency between imaging sessions. Calibrated weighted mean graylevels (WMGLs) for biological meterials had an average standard deviation of 5.9 graylevels (2.4% variation) during the course of the study. WMGLs for inorganic materials had an average standard deviation of 0.9 graylevels (0.4% variation). A trend towards increased image brightness, due to specimen and/or embedding media degradation, was observed in the biological tissues. No increase in rightness was observed for the inorgtanic specimens. Kurtosis and skewness tests revealed a slight deviation from normality in all specimens, which remained consistent between multiple imaging sessions. These results demonstrate the BSE image analysis of bones and mineral can be calibrated with negligible precision error allowing comparisons between data within and between multiple imaging sessions.

Normal motion of the lumbar spine as related to age and gender.

The CA-6000 Spine Motion Analyzer was used to measure the lumbar spine's range of motion (ROM). One hundred and four asymptomatic volunteers were examined to obtain normal values for flexion/extension, lateral bending, and axial rotation. A detailed error analysis was conducted to investigate the inter- and intraobserver reliability of the measurement equipment, the differences between passive and active examination, the effects of stretching exercises before examination, and the diurnal changes related to lumbar spine ROM. Subjects were divided into groups by age and gender. Values for each group were compared with respect to age and gender. The measurements were found to be consistent and repeatable. Stretching exercises were observed to increase ROM. Passive examination was recommended to achieve maximum ROM. ROM was observed to increase during the course of the day. A normative database was established showing significantly decreased motion as age increased, but no gender differences were discovered. The validity of the axial rotation values due to fixation difficulties is questioned.

Increased intracortical bone remodeling during lactation in beagle dogs.

There are substantial changes in skeletal and mineral metabolism during pregnancy and lactation. The purpose of this study was to determine the changes in intracortical bone remodeling and turnover during lactation in beagle dogs. A femur and rib were obtained from dogs near the end of lactation or soon after weaning and compared with nonlactating controls. Rib cortical bone had much higher bone turnover rates than did femoral diaphyseal cortical bone. The number of single-labeled osteons and the number of resorption spaces were significantly greater during lactation in both the rib and the femur. Additionally, the mineral apposition rate, basic multicellular unit activation frequency, and bone turnover rates were greater in the femoral cortical bone from the lactating dogs than from the controls. These data demonstrate that during lactation, intracortical bone remodeling increases, and this may provide a mechanism for the skeleton to be responsive to the calcium requirements of the mother. In addition, these data may help explain the transient decreases in cortical bone mineral density that are reported to occur during human lactation.

The contribution of cortical and cancellous bone to dual-energy X-ray absorptiometry measurements in the female proximal femur.

Dual-energy X-ray absorptiometry (DXA) is the most common method for determining bone mineral density (BMD) in the proximal femur. However, there remain questions concerning the contribution of cortical and cancellous bone to this technology in the proximal femur. The purpose of this investigation was to identify structural and compositional characteristics of human bone in the proximal femur that significantly influence DXA BMD measurements. Twenty-four femora were obtained at autopsy from Caucasian females ranging in age from 17 to 92 years (mean +/- SD, 61 +/- 25 years). DXA scans were performed on each specimen with a Hologic QDR-2000 densitometer. Direct measurements were determined from proximal femoral sections for cancellous bone (volume fraction, ash fraction, cancellous cross-sectional area and percent cancellous cross-sectional area), cortical bone (thickness, ash fraction, porosity, cortical cross-sectional area and percent cortical cross-sectional area) and anteroposterior thickness. These parameters were compared with the associated DXA measurements by means of simple and multiple regressions. Cancellous volume fraction was the best predictor of variability of DXA measurements for both the neck and trochanter, with an R2 of 0.87 and 0.76, respectively (p < 0.0001). There was only a minor influence of cortical factors such as thickness (neck and trochanter R2 = 0.51 and 0.42, respectively, p < 0.001) and trochanteric cross-sectional area (R2 = 0.21, p < 0.05). Although the accuracy for determining specific components of the proximal femur was low, the DXA BMD measurement was a strong predictor of cancellous bone factors, but not cortical bone factors that have been shown to change significantly with age.