Equivalence between adenosine and exercise thallium-201 myocardial tomography: a multicenter, prospective, crossover trial.

OBJECTIVES: The study was designed to compare pharmacologic and exercise stress during thallium-201 single-photon tomography in a multicenter prospective crossover trial. BACKGROUND: Both exercise and adenosine myocardial perfusion imaging have high sensitivity and specificity for detection of coronary artery disease. However, few data are available comparing these two stress tests in the same patients. METHODS: The study group consisted of 175 subjects: 55 healthy volunteers and 120 patients with suspected coronary artery disease. All subjects underwent two thallium tomographic tests performed 30 days apart, one during intravenous administration of adenosine (140 micrograms/kg per min for 6 min) and one during exercise stress. All images were computer quantified and interpreted without knowledge of the stress test performed. Interpretation agreement was assessed by kappa and Z statistics. RESULTS: Agreement on the presence of normal or abnormal tomograms by adenosine and exercise scintigraphy was 82.8% by visual analysis with kappa and Z statistics of 0.65 (p less than 0.0001) and 11.1 (p less than 0.00001), respectively. The agreement by computer quantification was 86% with kappa and Z statistics of 0.709 (p less than 0.0001) and 12.2 (p less than 0.00001), respectively. Agreement on localization of the perfusion defect to a specific coronary vascular territory varied from 82.7% to 91.4% with highly significant kappa and Z statistics (p less than 0.0001). There was a good correlation between quantified perfusion defect size by adenosine and exercise (r = 0.80, p less than 0.0001), but the values for defect size were significantly greater by adenosine scintigraphy (p = 0.0073). Adenosine side effects were frequent but transient and ceased spontaneously in most subjects within 1 to 2 min after the infusion was discontinued. CONCLUSIONS: Adenosine thallium-201 scintigraphy provides diagnostic information similar to that of exercise scintigraphy, although values for defect sizes are greater with adenosine.

Quantitative thallium-201 single-photon emission computed tomography during maximal pharmacologic coronary vasodilation with adenosine for assessing coronary artery disease.

The diagnostic value of maximal pharmacologic coronary vasodilation with intravenously administered adenosine in conjunction with thallium-201 single-photon emission computed tomography (SPECT) for detection of coronary artery disease was investigated in 101 consecutive patients who had concomitant coronary arteriography. Tomographic images were assessed visually and from computer-quantified polar maps of the thallium-201 distribution. Significant coronary artery disease, defined as greater than 50% luminal diameter stenosis, was present in 70 patients. The sensitivity for detecting patients with coronary artery disease using quantitative analysis was 87% in the total group, 82% in patients without myocardial infarction and 96% in those with prior myocardial infarction; the specificity was 90%. The sensitivity for diagnosing coronary artery disease in patients without infarction with single-, double-and triple-vessel disease was 76%, 86% and 90%, respectively. All individual stenoses were identified in 68% of patients with double-vessel disease and in 65% of those with triple-vessel disease. The extent of the perfusion defects, as quantified by polar maps, was directly related to the extent of coronary artery disease. In conclusion, quantitative thallium-201 SPECT during adenosine infusion has high sensitivity and specificity for diagnosing the presence of coronary artery disease, localizing the anatomic site of coronary stenosis and identifying the majority of affected vascular regions in patients with multivessel involvement.

The variable extent of jeopardized myocardium in patients with single vessel coronary artery disease: quantification by thallium-201 single photon emission computed tomography.

To assess the extent of jeopardized myocardium in patients with single vessel coronary artery disease of variable severity and location, quantitative exercise thallium-201 single photon emission computed tomography was performed in 158 consecutive patients with angiographically proved single vessel coronary artery disease. The extent of abnormal left ventricular perfusion was quantified from computer-generated polar maps of three-dimensional myocardial radioactivity. Patients with only a moderate (51% to 69%) stenosis tended to have a small perfusion defect irrespective of the coronary artery involved. Whereas a perfusion defect measuring greater than or equal to 10% of the left ventricle was found in 78% of patients with no prior infarction and severe (greater than or equal to 70%) stenosis, this was observed in only 24% of patients with moderate stenosis. Perfusion defect size increased with increasing severity of stenosis for the entire group without infarction and for those with left anterior descending, right and circumflex coronary artery stenosis. However, the correlation between stenosis severity and perfusion defect size was at best only modest (r = 0.38, p = 0.0001). The left anterior descending artery was shown to be the most important of the three coronary arteries for providing left ventricular perfusion. Proximal stenosis of this artery produced a perfusion defect approximately twice as large as that found in patients with a proximal right or circumflex artery stenosis. However, marked heterogeneity in perfusion defect size existed among all three vessels despite comparable stenosis severity. This was most apparent for the left anterior descending coronary artery, where mid vessel stenosis commonly produced a perfusion defect similar in size to that found in proximally stenosed vessels.(ABSTRACT TRUNCATED AT 250 WORDS)

Tolerance and safety of pharmacologic coronary vasodilation with adenosine in association with thallium-201 scintigraphy in patients with suspected coronary artery disease.

Adenosine thallium-201 myocardial scintigraphy is a promising test for coronary artery disease detection, but its safety has not been reported in large patient cohorts. Accordingly, the tolerance and safety profile of adenosine infusion were analyzed in 607 patients (351 men, 256 women, mean age 63 +/- 11 years) undergoing this test either because of suspected coronary artery disease (Group I, n = 482) or for risk stratification early (5.2 +/- 2.8 days) after myocardial infarction (Group II, n = 125). Adenosine increased the heart rate from 74.5 +/- 14.0 to 91.8 +/- 15.9 beats/min (p less than 0.001) and decreased systolic blood pressure from 137.8 +/- 26.8 to 120.7 +/- 26.1 mm Hg (p less than 0.001). Side effects were frequent and similar in both groups. Flushing occurred in 35%, chest pain in 34%, headache in 21% and dyspnea in 19% of patients. Only 35.6% of Group I patients with chest pain during adenosine infusion had concomitant transient perfusion abnormalities, compared with 60.7% of Group II patients (p less than 0.05). First- and second-degree AV block occurred in 9.6% and 3.6% of patients, respectively, and ischemic ST changes in 12.5% of cases. Concomitance of chest pain and ischemic ST depression was uncommon (6%) but, when present, predicted perfusion abnormalities in 73% of patients. Most side effects ceased rapidly after stopping the adenosine infusion. The side effects were severe in only 1.6% of patients and in only six patients (1%) was it necessary to discontinue the infusion. No serious adverse reactions such as acute myocardial infarction or death occurred.(ABSTRACT TRUNCATED AT 250 WORDS)

Quantitative exercise thallium-201 single photon emission computed tomography for the enhanced diagnosis of ischemic heart disease.

The clinical utility of exercise thallium-201 single photon emission computed tomography was investigated in 360 consecutive patients who had concomitant coronary arteriography. Tomographic images were assessed visually and from computer-quantified polar maps. Sensitivity for detecting coronary artery disease was comparably high using quantitative and visual analysis, although specificity tended to improve using the former method (87% versus 76%, p = 0.09). Quantitative analysis was superior to the visual method for identifying left anterior descending (81% versus 68%, p less than 0.05) and circumflex coronary artery (77% versus 60%, p less than 0.05) stenoses and detected most patients (92%) with multivessel coronary artery disease. Multivessel involvement was correctly predicted in 65% of the patients with more than one critically stenosed vessel. Exercise variables in patients with significant coronary artery disease were similar whether the tomographic images were normal or abnormal. However, patients with coronary stenoses and normal versus abnormal tomograms had a trend toward more single vessel disease (79% versus 62%, p = 0.07) and moderate coronary stenosis (66% versus 28%, p less than 0.001), but had less proximal left anterior descending artery involvement (8% versus 34%, p = 0.05). Computer-quantified perfusion defect size was directly related to the extent of coronary artery disease. Intra- and interobserver agreement for quantifying defects were excellent (r = 0.98 and 0.97, respectively). In conclusion, quantitative thallium-201 tomography offers improved detection of coronary artery disease, localization of the anatomic site of coronary stenosis, prediction of multivessel involvement and accurate determination of perfusion defect size, while maintaining a high specificity. Quantification of perfusion defects with single photon tomography may become important for assessing the effects of coronary reperfusion and prognostically stratifying patients with coronary artery disease.

Quantification of myocardial infarction during coronary occlusion and myocardial salvage after reperfusion using cardiac imaging with technetium-99m hexakis 2-methoxyisobutyl isonitrile.

Myocardial imaging with technetium-99m hexakis 2-methoxyisobutyl isonitrile was investigated as a means to assess myocardial infarct size during coronary occlusion and to quantify the extent of salvaged myocardium after coronary occlusion followed by reperfusion. Open chest dogs underwent either a permanent coronary artery occlusion (Group 1, n = 16) or a 2 h occlusion followed by reperfusion (Group 2, n = 15). Animals in both groups were killed 48 h after occlusion. During coronary occlusion, 23 of the 25 dogs that survived the coronary occlusions had abnormal myocardial scintigrams. The scintigraphic perfusion defect size correlated well with the pathologic infarct size (r = 0.85 and 0.95 by planar and tomographic imaging, respectively). The planar scintigraphic defect size, but not the tomographic defect size, overestimated the pathologic size. The planar scintigraphic defect size observed during coronary occlusion was markedly reduced 48 h after reperfusion (24.8 +/- 12.8% to 10.6 +/- 9.7% of the left ventricle, p less than 0.003). The uptake of technetium-99m hexakis 2-methoxyisobutyl isonitrile in the ischemic myocardium increased significantly 48 h after reperfusion (p less than 0.003) and correlated with the increase in regional myocardial blood flow, as assessed by radioactive microspheres (r = 0.83, p less than 0.01). Thus, myocardial imaging with technetium-99m hexakis 2-methoxyisobutyl isonitrile allows reliable demonstration of the presence of acute infarction, estimation of infarct size and quantification of the extent of salvaged myocardium after coronary reperfusion.

Mitochondrial DNA in the bark weevils: size, structure and heteroplasmy.

Mitochondrial DNA of higher animals has been described as an example of extreme efficiency in genome structure and function. Where exceptionally large size molecules have been found (greater than 20 kb), most have occurred as rare variants within a species, suggesting that these variants arise infrequently and do not persist for long periods in evolutionary time. In contrast, all individuals of at least three species of bark weevil (Curculionidae: Pissodes) possess a mitochondrial genome of unusually large size (30-36 kb). The molecule owes its large size to a dramatically enlarged A + T-rich region (9-13 kb). Gene content and order outside of this region appear to be identical to that found in Drosophila. A series of 0.8-2.0-kb repeated sequences occur adjacent to the large A + T rich region and have perhaps played a role in the generation of the large size as well as an unprecedented frequency of size variant heteroplasmy. Every weevil sampled in all three species (n = 219) exhibits anywhere from two to five distinct size classes of mtDNA. The persistence of this large amount of size polymorphism through two speciation events combined with the abundant size variation within individuals suggests that these molecules may not be subject to strong selection for small overall size and efficiency of replication. This pattern of variation contrasts strongly with the conservation of gene content and arrangement in the coding region of the molecule.

Cortical aging differences and fracture implications for the human femoral neck.

Clinical imaging techniques cannot consistently identify individuals at risk for hip fracture. Individual differences in falling likelihood partly account for these inconsistencies, but it is also thought that microscopic bone changes may play a role. In this study, subcapital, mid-neck, and trochanteric sites from eight young adult (26 +/- 7 years) and nine older (63 +/- 3 years) males were studied using backscattered electron imaging to identify age-related microscopic structural and mineral changes around the cortex. Cortical bone volume (BV(Ct)/TV), cortical void volume (Vd.V(Ct)/TV), hypermineralized bone volume (BV(H-min)/TV), the number of osteons/mm2 (N.On/B.Ar), lacunae/mm2 (N.Lc/B.Ar(Ct)) in the cortex, lacunae/mm2 (N.Lc/B.Ar(H-min)) in the hypermineralized phase, and cortical thickness (Ct.Th) were measured at subcapital, mid-neck, and trochanteric levels. Cortical void volume showed no differences (P = 0.26) between levels in the younger group, but differences (P < 0.05) were observed in the older group, indicating locational osteopenic differences. Cortical thickness differences were greater at the subcapital (27.7%) and mid-neck (25.2%) levels than at the trochanteric level (10.5%). Both age (P = 0.0022) and level-location interaction (P < 0.0001) influenced the hypermineralized bone volume present, with larger hypermineralized regions generally occurring at the thinner superior locations. Significant (P < 0.05) lacunar differences with aging in the hypermineralized phase suggest a necrotic origin. Artifactual cracks occurred preferentially within the hypermineralized phase, indicating localized reductions in fracture toughness, which may provide a site for crack initiation following an impact.

Aging changes in osteon mineralization in the human femoral neck.

Age-related mineral changes are associated with material property changes in cortical bone, which may affect its ability to transmit loads and resist fracture from falling. Aging changes in the mineral content of the femoral neck cortical bone are partly determined by osteonal remodeling events. The objective of this study was to measure mineral differences within the osteonal unit in the human femoral neck. In this study, cadaveric femora from male caucasians of two age groups (25 +/- 7, n = 6; 64 +/- 4, n = 6) were sectioned at the midfemoral neck. Using backscattered electron imaging, osteons from each of eight circumferential cortical regions were analyzed to determine osteonal mineral differences between regions, between the two age groups, and due to radial location within the osteons. Graylevel values from the backscattered electron imaging method were calibrated in equivalent ash content (wt%) units to provide a better understanding of the magnitude of differences observed. A pattern (p < 0.05) of decreasing osteonal mineralization was observed with distance from the central Haversian canal. Additionally, osteonal mineralization was highest in the superior (most lateral) regions of the femoral neck from both age groups, indicating that circumferential location is an influencing factor. The average overall equivalent ash content of osteons from the 17-35-yr group was 59.4 +/- 0.4% (mean +/- SE) by weight, while osteons of the group aged 60-71 measured 52.0 +/- 0.4% equivalent ash content, a decrease of 12.4%.(ABSTRACT TRUNCATED AT 250 WORDS)

Intracortical remodeling in adult rat long bones after fatigue loading.

Intracortical remodeling in the adult skeleton removes and replaces areas of compact bone that have sustained microdamage. Although studies have been performed in animal species in which there is an existing baseline of remodeling activity, laboratory rodents have been considered to have limited suitability as models for cortical bone turnover processes because of a lack of haversian remodeling activity. Supraphysiological cyclic axial loading of the ulna in vivo was used to induce bending with consequent fatigue and microdamage. Right ulnae of adult Sprague-Dawley rats were fatigue-loaded to a prefailure stopping point of 30% decrease in ulnae whole bone stiffness. Ten days after the first loading, left ulnae were fatigued in the same way. Ulnae were harvested immediately to allow comparison of the immediate response of the left ulna to the fatigue loads, and the biological response of the right leg to the fatigue challenge. Histomorphometry and confocal microscopy of basic fuchsin-stained bone sections were used to assess intracortical remodeling activity, microdamage, and osteocyte integrity. Bone microdamage (linear microcracks, as well as patches of diffuse basic fuchsin staining within the cortex) occurred in fatigue-loaded ulnar diaphyses. Ten days after fatigue loading, intracortical resorption was activated in ulnar cortices. Intracortical resorption occurred in preferential association with linear-type microcracks, with microcrack number density reduced almost 40% by 10 days after fatigue. Resorption spaces were also consistently observed within areas of the cortex in which no bone matrix damage could be detected. Confocal microscopy studies showed alterations of osteocyte and canalicular integrity around these resorption spaces. These studies reveal that: (1) rat bone undergoes intracortical remodeling in response to high levels of cyclic strain, which induce microdamage in the cortex; and (2) intracortical resorption is associated both with bone microdamage and with regions of altered osteocyte integrity. From these studies, we conclude that rats can initiate haversian remodeling in long bones in response to fatigue, and that osteocyte death or damage may provide one of the stimuli for this process.

First-pass radionuclide angiography using a multiwire gamma camera and tantalum-178.

A portable multiwire gamma camera (MWGC) with enhanced imaging characteristics relative to conventional sodium iodide camera has been evaluated with 178Ta, a short-lived, generator-produced radioisotope (half-life 9.3 min). First-pass radionuclide angiography (FPRA) was performed and results were compared to those obtained with FPRA using a multicrystal camera (MCC) and 99mTc in 38 patients. The overall left ventricular count sensitivity (counts/mCi/sec/millisteradians [msr]) was significantly higher with MWGC/178Ta (176 +/- 132 versus 108 +/- 49, p less than 0.001) yielding images of higher statistics with higher resolution collimation (31 versus 63 msr). Left ventricular ejection fraction was 0.54 +/- 0.18 by MWGC and 0.54 +/- 0.18 by MCC with an excellent correlation between the two techniques (r = 0.94, s.e.e. = 0.06). The detection of wall motion abnormality was virtually identical with the two techniques. Intra- and interobserver reproducibility by MWGC was excellent (r = 0.99 and 0.99, respectively). Thus, this new technology provides first-pass studies of higher statistical quality and improved resolution, affording more precise assessment of left ventricular performance and likelihood of further substantial improvement by use of even higher doses of 178Ta.

Molecular cloning of an alpha-esterase gene cluster on chromosome 3r of Drosophila melanogaster.

All or part of the alpha-esterase gene cluster in Drosophila melanogaster has been isolated by screening a YAC clone that spans cytological region 84D3-10 with consensus carboxyl/cholinesterase oligonucleotides. The cluster encompasses 11 putative esterase genes within 65 kb of genomic DNA and is one of the largest clusters of related protein-coding genes yet reported in Drosophila. The cluster must include the gene encoding the major alpha-esterase isozyme, EST9, which has previously been mapped to 84D3-5. It probably also includes the genes encoding the EST23, MCE and ALI esterases that have previously been mapped to 84D3-E2. The latter three are homologs of genes involved in organophosphate insecticide resistance in the sheep blowfly, Lucilia cuprina and the housefly, Musca domestica. Sequencing of one of the putative esterase genes in the Drosophila cluster, alpha E1, shows that it would encode features characteristic of an active carboxyl/cholinesterase, including the so-called catalytic triad, the nucleophilic elbow and oxyanion hole. It also shows that the closest relative of alpha E1 amongst previously published esterase sequences is ESTB1, which confers organophosphate resistance in Culex mosquitoes. We argue that we have cloned the D. melanogaster version of a major cluster of esterase genes which have variously mutated to confer organophosphate resistance in diverse Diptera.

Damage type and strain mode associations in human compact bone bending fatigue.

When compact bone is subjected to fatigue loading, it develops matrix microdamage, which reduces the tissue's ability to resist fracture. The relative influence of different strain modes on damage and strength in compact bone has not been characterized, to our knowledge. In this study, the nonuniform strain field produced by four-point bending was used to introduce fatigue damage into tibial bending beam specimens from men 40-49 years old. The specimens were then bulk-stained with basic fuchsin to mark damage surfaces and were examined histologically and with confocal microscopy to describe damage morphologies and position relative to tension and compression-strained regions of the specimen. Histomorphometric methods were used to quantify the amounts of different types of bone microdamage. Three major types were observed. In regions subjected to tensile strains, the bone had focal regions of diffusely increased basic fuchsin staining (i.e., diffuse microdamage). Confocal microscopy of these regions showed them to be composed of extensive networks of fine, ultrastructural-level cracks. In compressive strain regions, the tissue developed linear microcracks in interstitial areas similar to those originally described by Frost. Fine, tearing-type (wispy-appearing) cracks were observed near and in the plane of the neutral axis. The paths of these fine cracks were not influenced by microstructural boundaries. Other minor damage morphologies (sector-stained osteons, delamination of regions of lamellae, and intraosteonal cracking) were observed, but their distribution was unrelated to local strain field. Thus. in fatigue of human compact bone, the principal mechanisms of matrix failure (i.e., linear microcrack, diffuse damage foci, and tearing-type damage) are strongly dependent on local strain type.

Backscattered electron imaging: the role in calcified tissue and implant analysis.

The working distance and tilt studies helped to clarify the influences of specimen variability when the BSE mode is used in calcified tissue research. This work has shown that the BSEPs of cortical bone may be accurately maintained within 2 percent error over a 10 degree range of tilt, or 300 microns working distance variation. If future bone and implant investigators wish to conduct accurate, quantitative mineral microanalysis in bone, then standard grinding and polishing techniques should be adequate if calibration procedures are developed. The BSEP characteristics of the pure metals make them suitable to be used for calibrating the BSE signal. BSE analysis, with correlated biomechanical studies, will lead us to a better understanding of the relationships between structure, function, and mineral content in bone. On-line BSEP analysis techniques will expand our understanding of the mineralization events in bone which are associated with aging, weightlessness, pharmaceutical therapies, and the presence of biomaterials. The future of the BSE imaging technology and the contributions to be made in understanding the histometry, biomechanics and mineral content of bone as well as bone's response to implant materials has just begun to unfold.

Nucleotide variation at the hypervariable esterase 6 isozyme locus of Drosophila simulans.

Esterase 6 (Est-6/EST6) is polymorphic in both Drosophila melanogaster and D. simulans for two common allozyme forms, as well as for several other less common variants. Parallel latitudinal clines in the frequencies of the common EST6-F and EST6-S allozymes in these species have previously been interpreted in terms of a shared amino acid polymorphism that distinguishes the two variants and is subject to selection. Here we compare the sequences of four D. simulans Est-6 isolates and show that overall estimates of nucleotide heterozygosity in both coding and 5' flanking regions are more than threefold higher than those obtained previously for this gene in D. melanogaster. Nevertheless, the ratio of replacement to exon silent-site polymorphism in D. simulans is less than the ratio of replacement to silent divergence between D. simulans and D. melanogaster, which could be the result of increased efficiency of selection against replacement polymorphisms in D. simulans or to divergent selection between the two species. We also find that the amino acid polymorphisms separating EST6-F and EST6-S in D. simulans are not the same as those that separate these allozymes in D. melanogaster, implying that the shared clines do not reflect shared molecular targets for selection. All comparisons within and between the two species reveal a remarkable paucity of variation in a stretch of nearly 400 bp immediately 5' of the gene, indicative of strong selective constraint to retain essential aspects of Est-6 promoter function.

MCE activities and malathion resistances in field populations of the australian sheep blowfly (Lucilia cuprina).

Malathion resistance has been shown to be the result of a single point mutation in the LcalphaE7 gene in four independently isolated chromosomes of Lucilia cuprina. The resultant amino acid substitution specifies high malathion carboxylesterase (MCE) activity. We have assayed MCE activities and resistance to malathion in three sets of field-derived samples, two sets of isogenic lines and five mass populations, and show that resistance to malathion in these samples is associated with high MCE activity in both sets of isogenic lines and four of the five mass populations. Additional mechanisms contributing to MCE activity or malathion resistance may be present in one of the mass populations. A second point mutation in LcalphaE7 is responsible for conferring diazinon resistance by encoding an increased organophosphate (OP) hydrolase activity. We also assayed diazinon resistances from the same three samples and show that diazinon and malathion resistances were in complete disequilibrium, with two exceptions. One exception involves the mass population with additional resistance mechanism(s) and the other involves three isogenic lines that are resistant to both insecticides. The molecular data for these lines suggest that they carry a duplication of the LcalphaE7 gene.

Molecular insights into the evolution of an enzyme; esterase6 in Drosophila.

It is still a suspicion among some evolutionary biologists that the incursion of molecular biology into their field will do little more than determine the DNA sequence differences underlying evolutionary changes already evident at the organismal level. Work on an esterase enzyme involved in the reproductive biology of Drosophila belies this view. Although it is already one of the most intensively studied gene - enzyme systems at an organismal level, recent molecular invetigations reveal several unexpected, and, in some cases, still inexplicable phenomena in its evolutionary history.

Reproducible methods for calibrating the backscattered electron signal for quantitative assessment of mineral content in bone.

Backscattered electron (BSE) imaging shows promise for orthopaedic and bone research. BSE images of bone may be captured on-line directly from the scanning electron microscope (SEM), and then analyzed to produce a backscattered electron profile (BSEP), a modified image graylevel histogram which is representative of the mineral content in bone. The goals of this work were 1) develop a reproducible graylevel calibration technique for bone specimens, and 2) determine a conservative time interval during which SEM operating conditions would remain stable. Calibration standards containing pure aluminum and pure magnesium wires were placed in the SEM with human cancellous bone. Baseline imaging conditions were first established by adjusting the SEM until the bone image displayed good resolution and graylevel separation between regions of different mineral content. Microscope brightness and contrast controls were randomly changed to initiate the new operating conditions of another imaging session, and graylevel values from the calibration metals were used to readjust the microscope back to baseline operating conditions. Weighted mean graylevel values of the BSEPs from calibration trials were compared to those of the baseline. Data showed that bone images could be reproduced within 1.2 percent. It was also concluded that our equipment required calibration checks at 20 minute intervals.

Influence of mineral content and composition on graylevels in backscattered electron images of bone.

To determine the meaning of graylevels in backscattered electron (BSE) images of actual bone tissues, the influence of mineral content and mineral composition on BSE image graylevels was studied using chick bone tissue representing a broad age range. These tissues were analyzed for BSE image graylevels, Ca/P molar ratios, mineral composition mineral content (v/v), ash fraction (w/w), and density (g/cm3). Linear regression analyses showed that the weighted mean graylevels (WMGLs) in BSE images were positively correlated to ash fraction (r2 = 0.711), mineral content (r2 = 0.720), and density (r2 = 0.843). Although the Ca/P ratio increased from 1.65 in embryos to 1.80 in 2-year olds, the compositional changes corresponding to this Ca/P molar ratio were estimated to produce a relatively minor (< 4.0%) change in BSE image graylevel. These results demonstrate that graylevels in BSE images of actual bone tissue can be attributed to mineral content and density, but only as a coincidence of their association with atomic number.