A novel DXA-based hip failure index captures hip fragility independent of BMD.

Capability of a novel dual-energy X-ray absorptiometry (DXA)-based hip failure index (HiFI) to discriminate between hip fracture cases and controls was evaluated. Given the constraints of planar DXA, the femoral neck was assumed a foam-filled ( approximately trabecular bone), thin-walled ( approximately cortical bone) sandwich structure, while HiFI estimated the critical force sufficient to buckle the wall of such a structure. Proximal femur DXA data from 1379 women aged 65yr and older, 268 with prior hip fracture were used. Comparison between standard areal bone mineral density (BMD), femur strength index (FSI), and HiFI was based on areas under receiver operatoring characteristic curves (AUC). The mean femoral neck BMD (SD) was 0.689 (0.109) g/cm(2) among the cases and 0.768 (0.119) g/cm(2) among the controls; the mean FSI 1.33 (0.36) and 1.54 (0.41), and the mean HiFI -0.28 (0.14) and -0.18 (0.15), respectively; all intergroup differences were highly significant (p<0.001). The intergroup difference for HiFI remained significant (p<0.002) after adjusting for age and BMD or FSI. The AUCs were 0.696 (95% confidence interval [CI]: 0.661-0.730) for BMD, 0.665 (0.630-0.700) for FSI, and 0.701 (0.666-0.736) for HiFI. In conclusion, HiFI may capture structural traits that account for femoral neck fragility independently of BMD or FSI. Obviously, the use of actual geometric and structural information from three-dimensional imaging of the femoral neck would help diminish the crude assumptions of the present DXA approach and reveal the true potential of the HiFI approach to gauge hip fragility and identify at-risk individuals for hip fractures.

Precision of Lunar Achilles+ bone quality measurements: time dependency and multiple machine use in field studies.

Qualitative ultrasound (QUS) is a portable, safe and relatively inexpensive technique used to obtain information on bone mineral quality in adults and children. QUS measures bone stiffness index (SI) through the incorporation of speed of sound (SOS) and broadband ultrasound attenuation (BUA). QUS technology may prove to be extremely useful in field research where more than one machine is used over different periods of time. 13 adults (27.6+/-4.6 years old) were recruited to determine the internal stability of two Lunar Achilles+ QUS machines (Lunar1, Lunar2), as well as the repeatability in bone stiffness measures between the two machines over time. Triplicate measurements of the calcaneus were taken within the same day (n = 258) and at 1 week (n = 120), 6 months (n = 54) and 1 year (n = 18) apart to determine the time-dependent repeatability. Using paired t-tests and separate mixed effects models, there were no differences reported in SI, SOS or BUA values within one machine, or between two machines over these short- and long-term time-frames. These results indicate that QUS machines are internally consistent and different machines may be used over time to provide reliable measurements of changes in bone quality.

Femur strength index predicts hip fracture independent of bone density and hip axis length.

INTRODUCTION: Proximal femoral bone strength is not only a function of femoral bone mineral density (BMD), but also a function of the spatial distribution of bone mass intrinsic in structural geometric properties such as diameter, area, length, and angle of the femoral neck. Recent advancements in bone density measurement include software that can automatically calculate a variety of femoral structural variables that may be related to hip fracture risk. The purpose of this study was to compare femoral bone density, structure, and strength assessments obtained from dual-energy X-ray absorbtiometry (DXA) measurements in a group of women with and without hip fracture. METHODS: DXA measurements of the proximal femur were obtained from 2,506 women 50 years of age or older, 365 with prior hip fracture and 2,141 controls. In addition to the conventional densitometry measurements, structural variables were determined using the Hip Strength Analysis program, including hip axis length (HAL), cross-sectional moment of inertia (CSMI), and the femur strength index (FSI) calculated as the ratio of estimated compressive yield strength of the femoral neck to the expected compressive stress of a fall on the greater trochanter. RESULTS: Femoral neck BMD was significantly lower and HAL significantly higher in the fracture group compared with controls. Mean CSMI was not significantly different between fracture patients and controls after adjustment for BMD and HAL. FSI, after adjustment for T score and HAL, was significantly lower in the fracture group, consistent with a reduced capacity to withstand a fall without fracturing a hip. CONCLUSION: We conclude that BMD, HAL, and FSI are significant independent predictors of hip fracture.

Longitudinal quality control methodology for the quantitative ultrasound Achilles+ in clinical trial settings.

Appropriate quality assurance (QA) and quality control (QC) procedures have been well developed and validated for dual X-ray absorptiometry (DXA), and are widely applied in multicenter clinical trials to monitor device stability used to check the treatment effects on bone mineral density. This is not yet the case for quantitative ultrasound (QUS) technology, for which no QC approaches have yet been fully tested. The first Achilles (GE-Lunar Corporation, Madison, WI, USA) has been on the market for 10 years (1991). The goal of this study was to develop the QC methodology for the QUS Achilles+ device using its past/current experience (log and maintenance files.) as well as by integrating the progress made over the last years in the ultrasound domain so as to better understand the influence of temperature on ultrasound parameters. Because of the lack of confidence in the external black rubber phantom used in daily QC with the Achilles+, to monitor the device stability, we selected several QC parameters known to be influenced by potential malfunctions as experienced by the maintenance department of GE-Lunar company as well as the physical approach. These are phantom temperature-adjusted speed of sound (PSOS-TC) and broadband ultrasound attenuation (PBUA-TC), water speed of sound error (WSE), water spectrum slope (WSS) and water gain (WG). We used four Achilles+ devices perfectly stable during their entire QC range, to calculate the optimum thresholds (based mostly on 95% confidence interval) for each of these parameters as well as the precision for the in vitro SOS and BUA. An additional not fully stable Achilles device has been used to run a QC procedure example. The precision expressed as the CV was 0.22% and 0.65% for the PSOS-TC and PBUA-TC, respectively. The alarm thresholds used for QC process are +/- 0.6%, +/- 1.9%, +/- 6.8 m/s, +/- 5.3% and +/- 7.3% for the PSOS-TC, PBUA-TC, WSE, WSS and WG, respectively. Applying a logical approach on the impact of each parameter on each other as well as their respective reactivity to malfunctions, we build a QC process flowchart meant to detect real malfunction in the daily QC. We found that in case of real malfunctions, the in vivo SOS should be decreased by 1.33 m/s for each 1 m/s increase in WSE. Unfortunately, in vivo BUA can not be adjusted when real malfunction occurs. Nevertheless, the BUA can be qualified as bad quality data and excluded from the medical interpretation. Using the currently available phantom and parameters, the best possible QC procedures to detect long-term drift in the daily QC of the Achilles+ was developed. To fully validate our approach and gain confidence in the defined limits it is our plan to apply this QC processing to a higher number of QUS devices.

Culture confirmation of cytomegalovirus and herpes simplex virus by direct enzyme-labeled DNA probes and in situ hybridization.

Using probes consisting of horseradish peroxidase (HRP) directly attached to DNA, scrapings or trypsinized cells from 217 adequate clinical samples were cultured and analyzed in 3 blind studies by in situ hybridization for the presence of cytomegalovirus (CMV) and herpes simplex virus (HSV). Sixty samples were judged inadequate due to insufficient cell numbers; however, this problem was significantly decreased during the course of the study. One hundred and eighteen samples were found positive and 70 samples were found negative for CMV. Scrapings of cultured cells from 29 clinical samples revealed 9 samples which were positive and 20 samples which were negative for HSV. Forty-two additional samples, containing either uninfected cells or cells infected with various strains of CMV, were analyzed for the ability of the HRP-DNA CMV probe to detect such isolates. Twenty samples were positive and 22 negative for CMV. No false-negatives or false-positives were observed for either CMV or HSV. In addition to the specificity noted above neither the CMV nor the HSV DNA probe hybridized to potential contaminants found in clinical specimens.

Long term effects of peak strenuous effort on serum magnesium, lipids, and blood sugar in apparently healthy young men.

Earlier findings showed a sustained lowering of serum magnesium concentration (S-Mg) which indicated the presence of Mg deficit, and a parallel, delayed rise of blood sugar and serum lipids as a sequel to strenuous effort. S-Mg was still significantly decreased 3 months after termination of peak effort. To gain further perspective, we followed the biochemical sequels of exertion over an extended period of observation, while maintaining the same experimental conditions used earlier, which mimicked those employed in the training of military recruits. We examined two groups of military recruits, n = 15 (group 1), n = 16 (group 2), mean age 18.6, SD 1.3 and 18.7, SD 0.6, years respectively, who underwent a graded training programme of 7 months' duration culminating in a 120 km forced march. Blood was sampled for estimation of S-Mg in 20 soldiers on recruitment, 6 and 10 months after the 120 km march in group 1, and 9, 11 and 15 months after the march in group 2. Blood sugar and serum lipids were screened on recruitment and up to 11 months after the 120 km march. A significant lowering of mean S-Mg was found as late as 10 months after completion of the march in group 1, and 11 months in group 2 (P less than 0.01). Mean serum cholesterol and triglycerides showed a delayed rise, especially in group 2 (P less than 0.05 and P less than 0.001, respectively), whereas blood sugar decreased in group 1, but increased in group 2 (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Delayed metabolic changes after strenuous exertion in trained young men.

Twenty apparently healthy, young male volunteers, aged 18-25 (mean 19.3, SD 1.4) years received a 6 months standardized, graded outdoor physical training and were screened for serum magnesium concentration (S-Mg), serum calcium concentration (S-Ca), serum aspartate amino transferase (S-AST), serum alanine amino transferase (S-ALT), serum creatine kinase activity (S-CK), other laboratory variables, weight, and VO2 ml.kg-1.min-1 [corrected] (VO2 max), before a 70 km march, as well as at 1, 24 and 72 h and 18 days after. Maximal aerobic power, body weight, haemoglobin, haematocrit, serum creatinine, total protein and albumin remained unchanged throughout. Immediately after the march, S-Mg did not change, S-AST, S-ALT and S-CK rose, but the rise was not statistically significant, while small but significant rises in S-Ca (P less than 0.05, Student's t-test) and serum cholesterol (P less than 0.01) normalized at 24 h. At 72 h after the march, a significant fall in S-Mg was found (P less than 0.01), together with a second significant rise in S-Ca (P less than 0.05). After 18 days, with no intervening marches or dietary changes, S-Mg remained significantly lowered (P less than 0.05), mean S-ALT and S-CK became significantly raised for the first time (P less than 0.001 and P less than 0.01 respectively), whereas S-Ca normalized. Concomitantly, for the first time there was now a significant rise in blood sugar (P less than 0.001), serum triglycerides (P less than 0.01), and a second rise of serum cholesterol (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of denaturization on the immunogenicity and uveitogenicity of retinal S-antigen.

Previous studies have shown that alteration of pH or temperature of retina extract can affect its complement fixing reactivity with anti-S-antigen serum. To examine the effect of pH or heat on the immunogenicity and uveitogenicity of purified bovine S-antigen, guinea-pigs were injected with pH- or heat-treated S-antigen and evaluated for clinical and histopathological signs of uveoretinitis, histopathology of pineal gland, serum and intraocular S-antigen antibody reactivity, and S-antigen skin test reactivity. Guinea-pigs that received pH 7 or pH 10 treated S-antigen responded as did those that received untreated S-antigen. Guinea-pigs injected with pH 4 or heat-treated S-antigen exhibited lower incidence, later onset and less severe uveitis than those that received untreated S-antigen. Systemic responses of skin test reactivity to S-antigen were not different from those of the control group; pineal gland involvement and serum anti-S antibody reactivity were slightly reduced. Skin test responses of animals receiving treated antigen were less to the treated (injected) antigen than to untreated S-antigen. In addition, antibody responses of guinea-pigs receiving pH 4 or heat-treated antigen were less to the treated (injected) antigen than to untreated S-antigen. These results suggest that the sites on the S-antigen molecule responsible for various aspects of pathogenicity and immunogenicity do not have the same sensitivity to physical/chemical treatment and may reside on different parts of the molecule. Furthermore, the reactive sites especially for antibody and skin test reactivity, may be continuous sites.

Changes in serum magnesium concentration after strenuous exercise.

Serum magnesium concentration (S-Mg) was measured in 20 highly trained young men (mean age 19.5, +/- 0.5, range 18-20.5) before, and at 1 hour, 24 hours, 72 hours, and 3 months after a 120 km hike. As found in previous studies, S-Mg was significantly decreased at the end of the hike (p less than 0.001, [corrected] Student's t-test). In this group S-Mg had risen significantly after 24 hours in relation to the value at 1 hour (but not to starting value); yet, at 72 hours and 3 months later, it was once more significantly lower than the starting value (p less than 0.001 and p less than 0.05, respectively, Student's t-test). A marked elevation in serum creatine kinase activity (CK) suggests that the rise in S-Mg observed at 24 hours is the result of either exertional rhabdomyolysis or loss of membrane integrity, as a result of the strenuous exertion, since the CK had fallen sharply by 72 hours after the hike. The biphasic, statistically significant, lowering of S-Mg which persisted after 3 months suggests that strenuous exertion induces magnesium deficiency.

Measurement of magnesium in human tissues and fluids: a historical perspective.

The history of the development of methods to measure magnesium in biological systems is reviewed. The current optimal methods are discussed in detail as are some highly sensitive methods which may be used in special circumstances.

Retinal autoimmunity: two decades of research.

A brief review of our current knowledge of retinal autoimmunity and the pathogenesis of experimental autoimmune uveitis developed over the past two decades is presented. Attention is focused upon the discovery of retinal S-antigen, its isolation and characterization, manifestations of the disease produced, mechanisms of experimental autoimmune uveitis, analysis of the activities of the peptide fragments of the antigen, and potential relationships of the model to human ocular disease.

Rabbit ocular and pineal autoimmune response to retina antigens.

Different forms of experimental autoimmune uveitis (EAU) can be produced by varying protocols to present different autoantigens to several species of experimental animal. We have studied the clinical, histological and serological responses of rabbits to footpad injection of various fractions of retina extract. Rabbits injected with retina extract or S antigen developed posterior uveitis. However, rabbits injected with retina extract, also developed an anterior uveitis and pinealitis not seen in rabbits receiving S antigen. The Serological response of rabbits to retina extract was different than that to purified S antigen. Antisera of rabbits receiving retina extract reacted with rabbit retina and pineal gland as well as with guinea pig retina but not with guinea pig pineal gland. In contrast anti-S antigen sera reacted with rabbit retina and guinea pig retina and pineal gland but not with rabbit pineal gland. Gel filtration chromatography of the ammonium sulfate supernate of retina extract was used to differentiate the antigens with which these two sera reacted. An analysis of these experiments gives preliminary evidence of an autoantigen(s) of rabbit retina and pineal gland that is not S antigen. The existence of multiple autoantigens common to retina and pineal gland in various species is significant in that it further underscores the relationship of these tissues. Furthermore, it is not unrealistic to expect more than one autoantigen of retina or uvea to be involved in autoimmune uveitis.

Revisiting the Holy Cross football team hepatitis outbreak (1969) by serological analysis.

Clinical and biochemical data collected during the Holy Cross College football team hepatitis A outbreak in 1969 suggested that 32 team members had icteric hepatitis, 58 had anicteric illness, and only seven were not infected. Using a currently available radioimmunoassay, we tested stored serum samples obtained during the outbreak for IgM antibody to hepatitis A virus (IgM anti-HAV). Only individuals with icteric hepatitis were found to have IgM anti-HAV in serum; those with presumed anicteric illness were shown not to be infected with hepatitis A virus. The attack rate was thus only 34%, not 93% as originally reported, and the incidence of icteric illness in those infected was 100%, not 33%. This serological analysis of a classic outbreak of hepatitis A illustrates the utility and importance of IgM anti-HAV testing in seroepidemiologic investigations of hepatitis outbreaks.

Spinal motoneuron dendritic alteration after spinal cord hemisection in the rat.

Alteration of spinal motoneurons can be induced by axotomy, deafferentation, or deafferentation of spinal interneurons. In our experiment, dendritic profiles of rat lamina IX motoneurons were studied 1.5 to 2.5 mm rostral to a T2, T3 (interface) hemisection [six per group of normal, 7-, 14-, 30-, 60-, and 90-day postoperative (DPO) animals]. The spinal cord was impregnated using the Golgi technique and individual dendritic segments measured from coded slides, the data were computerized, and the length, number of segments, and dendritic profile were reconstructed. These data were heteroscedastic (alpha less than 0.005) making them inappropriate for ANOVA. However, the data did show two size classes of motoneurons at 14, 30, 60, and 90 DPO. At 7 DPO, there was a reduced dendritic field. At 14 to 60 DPO, there were gigantic motoneurons that were many times larger than normal or other neurons in these operated groups. At 90 DPO a large motoneuron was present. These data show that individual motoneurons have different regenerative dendritic responses after spinal injury.

Comparison of diluents for serum magnesium estimation by atomic absorption spectrophotometry.

In order to determine the optimal diluent for the determination of Mg by atomic absorption spectrophotometry, the following diluents were tested: deionized water, 0.1 mol/l HCl, 2.5 g/l SrCl2, 1.8 g/l LaCl3, 17.7 g/l LaCl3, half-saturated 8-hydroxyquinoline (oxine), 40 g/l trichloroacetic acid (TCA)/17.7 g/l LaCl3, and 100 g/l TCA/0.1 mol/l HCl. The calibration curves, obtained on the same day, were passed through the origin in the form of linear regressions. The variances about the calibration curves did not differ significantly (p greater than 0.05), while the calibration slopes for the diluents differed significantly (p less than 0.001). A standard serum solution was analyzed with the use of the eight diluents and the mean results did not differ significantly (p greater than 0.05). The % coefficient of variation varied from 0.8 to 2.0 and the percent recovery ranged from 95.5 to 102.5. The 99% confidence interval (CI) ranged from 0.02 to 0.09. Since conventional CIs do not take into account the random error of the calibration curve, the inverse CI were also calculated for each diluent. On both the conventional estimates and on the inverse CI, strontium chloride gave the best results, but only marginally in view of the interassay variability of approximately 1%.