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The use of a dissected bovine heart to teach cardiac sonography.

OBJECTIVES: To create and test a dissected bovine heart model (BHM) to facilitate the interpretation of cardiac sonography (CS). METHODS: After a pretest and an instructional video on CS, emergency physicians (EPs) were randomized into two groups. Group 1 viewed two-dimensional (2D) anatomic pictures of human hearts. Group 2 examined the BHM and the same anatomic pictures as group 1. The EPs retook the pretest. The differences between the raw pretest and posttest scores of the groups were compared with an unpaired Student's t-test. Multiple linear regression was used to adjust for confounding by variation in education and initial test scores. EPs with previous experience in CS were excluded from the analysis. RESULTS: Thirty-five participants met the inclusion criteria, 16 in group 1 and 19 in group 2. The groups were well balanced with respect to postgraduate year training. The EPs in group 1 had a higher average pretest score of 11.6 versus 8.1 in group 2. Compared with the pretest scores, the average improvements in group 1 and group 2 were 7.6 and 11.3 points, respectively. Group 2 improved an average of 3.7 points (95% confidence interval [95% CI] = 0.7 to 6.7; p = 0.016) more than group 1. After adjusting for confounding by the difference in initial scores, group 2 improved 1.8 (95% CI = -1.1 to 4.8; p = 0.22) more points on average than group 1. CONCLUSIONS: A dissected bovine heart model did not significantly improve the ability of EPs to label structures on static ultrasounds over inspection of static-labeled anatomic pictures alone.

Animals↗

Two-stage two-locus models in genome-wide association.

Studies in model organisms suggest that epistasis may play an important role in the etiology of complex diseases and traits in humans. With the era of large-scale genome-wide association studies fast approaching, it is important to quantify whether it will be possible to detect interacting loci using realistic sample sizes in humans and to what extent undetected epistasis will adversely affect power to detect association when single-locus approaches are employed. We therefore investigated the power to detect association for an extensive range of two-locus quantitative trait models that incorporated varying degrees of epistasis. We compared the power to detect association using a single-locus model that ignored interaction effects, a full two-locus model that allowed for interactions, and, most important, two two-stage strategies whereby a subset of loci initially identified using single-locus tests were analyzed using the full two-locus model. Despite the penalty introduced by multiple testing, fitting the full two-locus model performed better than single-locus tests for many of the situations considered, particularly when compared with attempts to detect both individual loci. Using a two-stage strategy reduced the computational burden associated with performing an exhaustive two-locus search across the genome but was not as powerful as the exhaustive search when loci interacted. Two-stage approaches also increased the risk of missing interacting loci that contributed little effect at the margins. Based on our extensive simulations, our results suggest that an exhaustive search involving all pairwise combinations of markers across the genome might provide a useful complement to single-locus scans in identifying interacting loci that contribute to moderate proportions of the phenotypic variance.

Alleles↗

Angiogenesis assays: a critical overview.

BACKGROUND: Angiogenesis, the formation of new blood vessels, is an integral part of both normal developmental processes and numerous pathologies, ranging from tumor growth and metastasis to inflammation and ocular disease. Angiogenesis assays are used to test efficacy of both pro- and antiangiogenic agents. METHODS: Most studies of angiogenesis inducers and inhibitors rely on various models, both in vitro and in vivo, as indicators of efficacy. In this report we describe the principal methods now in use: the in vivo Matrigel plug and corneal neovascularization assays, the in vivo/in vitro chick chorioallantoic membrane (CAM) assay, and the in vitro cellular (proliferation, migration, tube formation) and organotypic (aortic ring) assays. We include description of two new methods, the chick aortic arch and the Matrigel sponge assays. CONCLUSIONS: In vitro tests are valuable, can be carried out expeditiously, and lend themselves to quantification, but must be interpreted with extreme caution. In vitro tests are best viewed as providing initial information, subject to confirmation by in vivo assays. Multiple tests should be used to obtain maximum benefit from in vitro tests. In vivo tests are more difficult and time-consuming to perform, thereby limiting the number of tests that can run at any one time. Quantification is generally more difficult as well. However, in vivo assays are essential because of the complex nature of vascular responses to test reagents, responses that no in vitro model can fully achieve.

Animals↗

The interplay between emotional and cognitive recovery after closed head injury.

This study examined 59 patients who had suffered closed head injuries with respect to their MMPI scores and Category Test scores as assessed shortly post-injury and on subsequent assessment. The number of MMPI scales elevated above t = 70 on the first assessment was negatively correlated with the magnitude of improvement made on the Category Test. Multiple regression analyses showed that the extent of cognitive deficit, as indicated by the Category Test, could be predicted from the degree of psychopathology, with MMPI scales 2 (depression), 7 (psychasthenia) and 8 (schizophrenia) being highly predictive of Category Test performance. These results suggest that patients with better psychological functioning perform at a higher level and make a greater post-traumatic recovery on neurocognitive tests. Although we cannot determine if the emotional impairment is due to neuropathology or is reactive in nature, there are clear implications for rehabilitation.

Adult↗

Dyslipoproteinemias and manifestations of coronary heart disease. The Lipid Research Clinics Program Prevalence Study.

The Lipid Research Clinics Prevalence Study population underwent a noninvasive evaluation to assess the odds of coronary heart disease (CHD) among individuals with various types of dyslipoproteinemia (DLP) relative to the odds for individuals free of DLP in a nonselected population. The noninvasive evaluation included the Rose questionnaire for angina, a resting electrocardiogram, and a graded treadmill exercise test. The presence of manifestations of CHD was assessed by a combination of these findings. Multiple linear regression and multiple logistic regression analyses were used to evaluate associations between CHD and DLP, with adjustments for the following covariates: age, body mass, systolic blood pressure, alcohol consumption, and cigarette use. There were no consistent associations in women or in men 30 to 49 years old. In men 50 to 69 years old, the mean high-density lipoprotein cholesterol (HDL-C) values and HDL-C/cholesterol ratio were significantly lower in the definite CHD and angina categories, compared with the negative (no evidence of CHD) group. Higher odds ratios for CHD were present in those classified as having type IIa, type IV, and hypoHDL DLPs, relative to those classified as normal by the phenotyping algorithm. These associations were consistent across the several categories of CHD manifestations, but did not reach statistical significance after adjustment for multiple testing. A statistically significant lower odds ratio for CHD was observed in men 50 to 69 classified as having hyperHDL when compared with those without DLPs. The associations between CHD and the various forms of DLP were consistent with those observed between CHD and the plasma lipid and lipoprotein-cholesterol concentrations. The Lipid Research Clinics Prevalence Study confirmed the relationship between elevated low-density lipoprotein cholesterol, decreased HDL-C, and noninvasively determined CHD in a free-living asymptomatic population of older men.

Adult↗

Evaluation of an agar gel immunodiffusion test kit for detection of antibodies to Mycobacterium paratuberculosis in sheep.

OBJECTIVE: To determine whether a commercially available agar gel immunodiffusion test approved for detecting antibodies to Mycobacterium paratuberculosis in cattle could be used for sheep. DESIGN: Experimental trial. SAMPLE POPULATION: Serum samples from 27 sheep confirmed to have paratuberculosis by means of acid-fast staining of smears of ileal mucosa, histologic examination of tissues, or bacteriologic culture; 7 sheep with clinical signs of paratuberculosis; and 55 sheep from 5 uninfected flocks. PROCEDURE: Serum samples were tested concurrently with the commercially available test and with a previously validated agar gel immunodiffusion test. Multiple samples collected from 13 infected sheep over a period of 6 years were also tested so that each test's ability to detect onset of seropositivity could be compared. RESULTS: For both tests, results for samples from all 55 uninfected sheep were negative, results for samples from 32 of the 34 sheep with paratuberculosis were positive, and results for the remaining 2 sheep with paratuberculosis were negative. Results of both tests were in agreement for 50 of 54 samples obtained from 13 infected sheep over time. The 4 samples for which results of the 2 tests disagreed were the fourth, eighth, and ninth of 10 samples from 1 sheep and the first of 6 samples from a second sheep. For all 4 samples, the commercially available assay yielded a weak-positive result, but the previously described test yielded a negative result. CLINICAL IMPLICATIONS: The commercially available agar gel immunodiffusion test approved for use in cattle may be useful in the differential diagnosis of paratuberculosis in sheep.

Animals↗

Multivariate approach to quality control in clinical chemistry.

When monitoring analyzer performance in the clinical setting, laboratories are required to test multiple concentrations of control material on a daily basis. Because of the nature of laboratory testing, there is the potential for correlation between the concentrations of control material being monitored. Although traditional clinical quality-control approaches make an underlying assumption of independence with respect to the control concentrations, this will not always be the case. The presence of correlation in some circumstances suggests the use of a new approach for evaluating clinical laboratory monitoring data: the multivariate control chart. Such a chart (the chi2 chart) is evaluated and compared with traditional quality-control approaches used in the laboratory setting. Results indicate that the multivariate approach provides an attractive alternative to many traditional methods of quality assurance when control concentrations are correlated.

Chemistry, Clinical↗

WAIS and neuropsychological tests: common and unique variance within an epileptic population.

Neuropsychological assessment batteries, which traditionally require lengthy administration times, may be composed of multiple tests which measure essentially the same performance attributes. Test redundancies of this nature compromise the adequacy of a test battery from a cost containment perspective. The present study evaluated the common and unique variance of the WAIS relative to a modified and expanded Halstead-Reitan test battery to determine if information obtained from the WAIS is sufficiently nonredundant to justify its inclusion as part of this assessment battery. Cross-validated estimates of bivariate and multivariate relationships between these instruments were generated from two samples (n = 125) of epileptics. Results indicated statistically significant redundancies, yet, when interpreted from a clinical perspective it was evident that neuropsychological tests could not accurately duplicate information provided by the WAIS. Accordingly, it was not recommended that the WAIS be eliminated from this test battery in order to reduce test administration time.

Adolescent↗

Rank-based tests for dose finding in nonmonotonic dose-response settings.

Lim and Wolfe (1997, Biometrics 53, 410-418) proposed rank-based multiple test procedures for identifying the dose levels that are more effective than the zero-dose control in randomized complete block designs when it can be assumed that the efficacy of the increasing dose levels is monotonically increasing up to a point, followed by a monotonic decrease. Modifications of the Lim-Wolfe tests are suggested that provide more practical and powerful alternatives. Two numerical examples are illustrated and the results of a Monte Carlo power study are presented.

Animals↗

Power comparisons between the TDT and two likelihood-based methods.

We compare the statistical power of the transmission disequilibrium test (TDT) with that of two likelihood-based linkage tests, the classical LOD score and a modified LOD score in which a linkage disequilibrium (LD) parameter is incorporated into the likelihood (LD-LOD). We hypothesize that, when LD is present, the LD-LOD will have the greatest power of the three tests because the TDT breaks a multiplex pedigree into triads, and the LOD score has previously been shown to have lower power when LD is present but not accounted for. We test this hypothesis using a simulation study in which we generate affected sib-pair (ASP) pedigrees under a range of genetic models, varying the genotypic relative risk (GRR) from 6 to 16. Because the likelihood-based tests require that a genetic model be specified, we compare the tests under two scenarios. First, we assume the true genetic model in the analysis, and second, we compare the tests when the LD-LOD (LOD) is maximized over two wrong genetic models. For the generating models we considered, we find that the LD-LOD has greater power than the TDT even when the genetic models is mis-specified and the results corrected for multiple tests. Extreme differences occur under the multiplicative and dominant models, for which the difference in power is as high as 40% at complete LD. The LOD score provides the lowest power in the presence of LD for the range of GRR considered here.

Adult↗

Equating and item banking with the Rasch model.

This article describes Rasch measurement procedures for equating multiple test forms or calibrating an item bank. The procedures entail (a) selecting an appropriate data collection design, (b) estimating parameters, (c) transforming the parameters from multiple forms to a common scale, and (d) evaluating the quality of the linkage between these forms. Data collection designs include (a) anchor tests, (b) single group, (c) single data set, and (d) equivalent groups. Estimation procedures may involve (a) separate or (b) simultaneous calibration of data from multiple forms. Transformation is typically accomplished using (a) estimation scaling, but may involve (b) parameter anchoring or (c) computing equating constants. Link quality is evaluated using four fit indices: (a) item-within-link, (b) item-between-link, (c) link-within-bank, and (d) form-within-bank. These procedures are illustrated using an anchor test design.

Data Collection↗

AAEM minimonograph #48: autonomic nervous system testing.

The autonomic nervous system maintains internal homeostasis by regulating cardiovascular, thermoregulatory, gastrointestinal, genitourinary, exocrine, and pupillary function. Testing and quantifying autonomic nervous system function is an important but difficult area of clinical neurophysiology. Tests of parasympathetic cardiovagal regulation include heart rate analysis during standing (the 30:15 ratio), heart rate variation with deep breathing, and the Valsalva ratio. Tests of sympathetic adrenergic vascular regulation include blood pressure analysis while standing, the Valsalva maneuver, sustained handgrip, mental stress, and cold water immersion. Tests of sympathetic cholinergic sudomotor function include the sympathetic skin response, quantitative sudomotor axon reflex test, sweat box testing, and quantification of sweat imprints. Pupil function is tested pharmacologically and with pupiilographic techniques. Tests of gastrointestinal and genitourinary function do not satisfactorily isolate autonomic regulation from their other functions. The available tests have various sensitivities and ease of administration. They are typically administered in a battery of multiple tests, which improves sensitivity and reliability, and allows probing of various autonomic functions.

Autonomic Nervous System↗

No evidence for cognitive side effects after 6 months of vagus nerve stimulation in epilepsy patients.

Vagus nerve stimulation (VNS) can reduce seizure frequency in epilepsy patients and may affect central mechanisms of brain functioning. Experimental studies have provided evidence of cognitive alterations during VNS on phases. This single-arm follow-up study evaluates the potential of VNS to affect cognitive performance following long-term treatment. Thirty-six adult patients with medication-resistant epilepsies enrolled. Cognition was assessed before and at least 6 months after implantation of the stimulation device by a comprehensive neuropsychological assessment battery comprising tests on attention, motor functioning, short-term memory, learning and memory, and executive functions. Neither multiple testing of single score changes nor multivariate testing of cognitive domains revealed significant pre-post changes. Improvements in attentional performance were completely explained by practice effects as is usually expected. In particular, no negative side effects were revealed. These findings are in line with the clinical impression that VNS does not affect cognitive performance.

Journal Article↗

Longitudinal analysis of haplotypes and polymorphisms of the APOA5 and APOC3 genes associated with variation in serum triglyceride levels: the Bogalusa Heart Study.

Polymorphisms in the APOC3 and APOA5 genes, from the APOA1/APOC3/APOA4/APOA5 gene cluster on chromosome 11q23, have been associated with interindividual variation in plasma triglycerides. APOA5 polymorphisms implicated include 2 in the promoter region (-1131 T/C and -3 A/G) and 1 in exon 2 (+56 C/G). APOC3 polymorphisms implicated include 1 (SstI) in the 3' untranslated region and 1 (-2854 G/T) in the APOC3-APOA4 intergenic region. We analyzed the associations of haplotypes and multilocus genotypes of these polymorphisms on longitudinal serum triglyceride profiles in 360 African American and 823 white subjects from the Bogalusa Heart Study. Subjects were examined from 2 to 8 times (mean +/- SD, 5.4 +/- 1.3) between 1973 and 1996, at ages ranging from 4 to 38 years, with 1978 observations in African Americans and 4465 in whites. Serum triglycerides were significantly higher among whites across all ages. Allele frequencies differed significantly between African Americans and whites at all but the APOA5 +56 C/G locus. Linkage disequilibrium among the loci was higher in whites and haplotype diversity lower: 6 haplotypes had estimated frequencies of more than 1% in African Americans, 5 in whites. Individually, all polymorphisms except APOC3 -2854 G/T showed significant associations with triglyceride levels in the full sample. However, genotype models including all 5 loci showed significant triglyceride associations for only 3 (APOC3 SstI, APOA5 -1131 T/C, and APOA5 +56 C/G); significant interactions among them indicated their effects were not independent. Neither APOC3 -2854 G/T nor APOA5 -3 A/G had significant effects when the other 3 loci were in the models. The EM algorithm was used to estimate haplotype frequencies and assign haplotype probabilities to individuals, which is conditional on their genotypes; individuals' haplotype probability vectors were then used as predictors in multilevel mixed models of longitudinal triglyceride profiles. Of haplotypes comprising, in order, APOC3 SstI and -2854 G/T and APOA5 -1131 T/C, -3 A/G, and +56 C/G, 3 were significantly associated with higher triglycerides, even after adjusting for multiple tests: GGTAG (P = .002), GTTAG (P < .0001), and CGCGC (P = .0002). Each GGTAG haplotype carried would be expected to raise triglyceride levels (relative to those of GTTAC homozygotes) by approximately 19 mg/dL, each GTTAG haplotype by approximately 15 mg/dL, and each CGCGC haplotype by approximately 7 mg/dL. Haplotypes comprising the 3 loci implicated by genotype analyses (SstI, -1131 T/C, and +56 C/G) were also tested: haplotypes C_C_C and G_T_G significantly raised triglycerides, even after adjustment for multiple comparisons (P < .002 for both), with each copy of C_C_C expected to raise triglycerides by approximately 7 mg/dL and each copy of G_T_G by approximately 15 mg/dL. Overall, our findings support those of others in associating specific polymorphisms and haplotypes in the APOA1/C3/A4/A5 gene cluster with higher serum triglyceride levels. However, the degree to which polymorphisms in the APOC3 and APOA5 genes may be independently associated with triglyceride levels remains to be determined.

Adolescent↗

In vitro remineralisation of eroded enamel lesions by saliva.

OBJECTIVES: It is speculated that saliva, with its mineral content, may possess a reparative effect on an early erosion which is characterised by softened surface and slight subsurface demineralisation in addition to a crater. This study aimed to determine the possible remineralisation of early enamel erosion by saliva. METHODS: Eroded lesions were produced in bovine incisors by 1-h immersion in orange juice. Control sections and three experimental slabs were produced from each tooth. The three slabs were assigned randomly to one of three remineralising agents: clarified natural saliva (NS), artificial saliva (AS) and remineralising solution (RS). All solutions had a pH of 7.2, a fluoride concentration of 0.022 ppm, and were changed daily. NS was collected daily from the same individual at the same time of day. The specimens were exposed to their respective remineralising agents for 28 days. Using microradiography and image analysis, the mineral loss (Delta z) and lesion depth (ld) were quantified in sections cut from the control and experimental slabs. RESULTS: A significant (p<0.001) amount of mineral was gained following exposure to each remineralising agent. Significantly less Delta z and ld were observed for the experimental groups compared with the control group (p<0.001; paired t-test). This effect was greatest with RS and least with AS. Inter-group comparison (Duncan multiple tests) showed no significant difference in Delta z among the experimental groups, however ld was significantly higher for AS (p<0.001) compared with RS and NS, and no difference was observed between RS and NS. CONCLUSION: Saliva as well as remineralising solutions can remineralise early enamel erosion.

Animals↗

A feasibility study of quantitative respirator fit testing by controlled negative pressure.

The feasibility of using a direct measure of respirator leakage flow rate as a quantitative index of respirator face seal fit has been explored through the use of a new controlled negative pressure method. The method is based on exhausting air from a temporarily sealed respirator facepiece at a rate sufficient to generate and then sustain a constant negative pressure inside the facepiece while the wearer holds his breath. The magnitude of the negative pressure is preselected to replicate the mean inspiratory pressure inside the mask during normal wear. With the air-purifying paths into the respirator temporarily blocked, measurement of the exhaust flow rate yields a synonymous measure of the leakage flow rate into the mask during inspiration under normal use conditions. The feasibility of using the new method to quantify respirator fit was assessed in a preliminary study that compared its performance with a quantitative fit test method based on the use of dichlorodifluoromethane as a challenge agent. Study data exhibit a high degree of correlation (r greater than 0.99) and no significant difference between the two methods over a range of controlled mask leakage rates. A major advantage of the new method is that a worker can be fit tested with his assigned respirator because the method does not require a destructive sampling probe. Other significant benefits compared to current methods used to quantify respirator fit appear to include (1) ease of test administration, (2) simplicity of test components, (3) lack of a potentially toxic challenge agent, (4) a straightforward calibration procedure, (5) multiple test capability, (6) immediacy of test results, and (7) field portability of the test system.

Air Pollutants↗

Inflammatory cell types and clinical features of interstitial cystitis.

PURPOSE: We tested whether the types of inflammatory cells seen on bladder biopsies were associated with other clinical features and urinary markers of interstitial cystitis. MATERIALS AND METHODS: Bladder biopsies from 30 interstitial cystitis patients were evaluated by immunohistochemical staining for T cells, B cells, macrophages and human leukocyte antigen-DR positive cells. These findings were tested for associations with clinical features and urinary markers of interstitial cystitis using alpha = 0.01 because multiple tests were performed. RESULTS: Overall severity of inflammation was significantly associated with age at symptom onset, symptom relief after bladder distention and urinary interleukin-6 levels. Patients with severe inflammation had trends toward smaller bladder capacity under anesthesia, increased bladder vascularity and mucosal cracks, lower urinary MUC-1 glycoprotein levels and absence of bloating as a symptom. B cell staining was significantly associated with severe inflammation, symptom relief after distention and absence of bloating as a symptom. T cell staining was significantly associated with severe inflammation and age at symptom onset. Human leukocyte antigen-DR staining had trends with symptoms, including presence of bloating, constant urge to void and absence of burning. Macrophage staining did not associate with any features tested at the alpha = 0.05 level. CONCLUSIONS: Interstitial cystitis patients with severe inflammation have different age, treatment response and urinary marker levels than those with mild inflammation. These findings suggest that the 2 patient groups have different underlying pathophysiologies. The significant associations for T and B cell staining were similar to those for overall inflammation.

B-Lymphocytes↗

Effects of selection in a prospective study of forced expiratory volume in Vermont granite workers.

Granite workers lost to follow-up during a 5-yr study of pulmonary function were examined. We found that in workers who had left the industry for other employment, forced expiratory volume in one second (FEV1) values declined at a faster rate than in those who remained employed or retired during the study period. We investigated the application of a currently recommended repeatability criterion for pulmonary function tests by looking at different patterns of failing to satisfy the criterion (PFT-failure) over multiple testing sessions. Workers with persistent PFT-failure were found to have faster rates of FEV1 decline than did subjects with none or only intermittent failures. In a regression analysis, job termination was found to be more strongly associated with a steep FEV1 slope when it had been preceded by a pattern of persistent PFT-failure.

Adult↗