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Regulation of gene expression in intervertebral disc cells by low and high hydrostatic pressure.

Intervertebral disc structures are exposed to wide ranges of intradiscal hydrostatic pressure during different loading exercises and are at their minimum during lying or relaxed sitting and at maximum during lifting weights with a round back. We hypothesize that these different loading magnitudes influence the intervertebral disc (IVD) by alteration of disc matrix turnover depending on their magnitudes. Therefore the aim of this study was to assess changes in gene expression of human nucleus cells after the application of low hydrostatic pressure (0.25 MPa) and high hydrostatic pressure (2.5 MPa). IVD cells isolated from the nucleus of human (n = 18) and bovine (n = 24 from four animals) disc biopsies were seeded into three-dimensional collagen type-I matrices and exposed to the different loading magnitudes by specially developed pressure chambers. The lower pressure range (0.25 MPa, 30 min, 0.1 Hz) was applied with a recently published device by using an external compression cylinder. For the application of higher loads (2.5 MPa, 30 min, 0.1 Hz) the cell-loaded collagen gels were sealed into sterile bags with culture medium and stimulated in a newly developed water-filled compression cylinder by using a loading frame. These methods allowed the comparison of loading regimes in a wide physiological range under an equal three-dimensional culture conditions. Cells were harvested 24 h after the end of stimulation and changes in the expression of genes known to influence IVD matrix turnover (collagen-I, collagen-II, aggrecan, MMP1, MMP2, MMP3, MMP13) were analyzed by real-time RT-PCR. A Wilcoxon signed-rank test(1) and a Wilcoxon 2-sample test(2) were performed to detect differences between the stimulated and control samples(1) and differences between low and high hydrostatic pressure(2). Multiple testing was considered by adjusting the p value appropriately. Both regimes of hydrostatic pressure influenced gene expression in nucleus cells with opposite tendencies for the matrix forming proteins aggrecan and collagen type-I in response to the two different pressure magnitudes: Low hydrostatic-pressure (0.25 MPa) tended to increase collagen-I and aggrecan expression of human nucleus cells (P < 0.05) but only to a small degree. High hydrostatic pressure (2.5 MPa) tended to decrease gene expression of all anabolic proteins with significant effects on aggrecan expression of nucleus cells (P = 0.004). Low hydrostatic pressure had no influence on the expression of matrix metalloproteinases (MMP1, MMP2, MMP3 and MMP13). In contrast, high hydrostatic pressure tended to increase the expression of MMP1, MMP3 and MMP13 of human nucleus cells with high individual-individual variations. The decreased expression of aggrecan (P = 0.008) and collagen type II (P = 0.023) and the increased MMP3 expression (P = 0.008) in response to high hydrostatic pressure could be confirmed in additional experiments with bovine nucleus cells. These results suggest that hydrostatic pressure as one of the physiological stimuli of the IVD may influence matrix turnover in a magnitude dependent way. Low hydrostatic pressure (0.25 MPa) has quite small influences with a tendency to anabolic effects, whereas high hydrostatic pressure (2.5 MPa) tends to decrease the matrix protein expression with a tendency to increase some matrix-turnover enzymes. Therefore, hydrostatic pressure may regulate disc matrix turnover in a dose-dependent way.

Adolescent↗

The significance of HPA axis disturbance in panic disorder.

Agoraphobic and panic disorder patients underwent 1-mg Dexamethasone Suppression Tests (DST) before, during, and after an 8-week trial of diazepam, alprazolam, or placebo. Previously described, never-ill controls underwent similar testing. At baseline, 21 of 82 (25.6%) panic disorder and 5 of 38 (13.2%) controls were nonsuppressors. This difference grew more marked with multiple testing over a 2-month period; 18 of 44 (40.9%) panic disorder patients were nonsuppressors on at least 1 of 3 tests compared with only 5 of 35 (14.3%) controls (p = 0.006). DST results were related to severity, but not to the presence or absence, of depressive syndromes. Control for plasma dexamethasone levels left highly significant differences in postdexamethasone cortisol across diagnostic groups. Neither DST results nor plasma dexamethasone levels changed in concert with clinical change, and type of treatment had little differential effect on these measures. Nor did DST results predict subsequent course when active treatment was extended by 6 months. However, DST results during the initial 8 weeks of treatment were strongly related to relapse when medications were tapered, even though this occurred 6 months after the last DST.

Adult↗

Clinical tests of the sacroiliac joint. A systematic methodological review. Part 1: Reliability.

In the literature concerning the sacroiliac joint (SIJ) there are numerous specific tests used to detect joint mobility or pain provocation. In this article the authors have reviewed 11 studies which investigated the reliability of these tests. The methodological quality of the studies was tested by a list of criteria developed by the authors. This list consisted of three categories: (1) study population, (2) test procedures and (3) test results. To each criterion a weighting was attached. The methodological score for nine out of the 11 studies was found to be acceptable. The results of this review, however, could not demonstrate reliable outcomes and therefore no evidence on which to base acceptance of mobility tests of the SIJ into daily clinical practice. There are no indications that 'upgrading' of methodological quality would have improved the final conclusions. With respect to pain provocation tests, the findings did not show the same trend. Two studies demonstrated reliable results using the Gaenslen test and the Thigh thrust test. One study showed acceptable reliability for five other pain provocation tests; however, since other authors have described contradictory results, there is a necessity for further research in this area with an emphasis on multiple test scores and pain provocation tests of the SIJ.

Humans↗

A comparative study of tests of cardiac parasympathetic nervous activity in healthy human subjects.

Conventional tests of cardiac parasympathetic nervous system (PNS) activity were compared with the high frequency component of the heart rate power spectrum in a heterogenous group of normal subjects encompassing a wide age range. The data suggest that the linear associations between the various conventional tests of parasympathetic nervous activity even when statistically significant, were relatively modest, with r values ranging from 0.23 to 0.53. Three of the five conventional tests of parasympathetic nervous activity were significantly correlated (r = 0.33 to 0.46) with the absolute high frequency power of RR variability. However, these relationships were poorer and non-significant when the high frequency power spectrum was normalized for total power (r = 0.06 to 0.19, NS). An evaluation of the heart rate responses to cough and a single maximal hand grip indicated that the responses were repeatable but that the extent to which these manoeuvres induced vagally mediated cardioacceleration was significantly lesser than the other conventional tests of PNS activity. Taken together, the data suggest that despite the advent of heart rate variability measures, it is advisable to use multiple tests of parasympathetic nervous activity while evaluating autonomic dysfunction, since, despite the specificity of the tests, there is a variable correlation between them.

Adolescent↗

An empirical Bayes approach to inferring large-scale gene association networks.

MOTIVATION: Genetic networks are often described statistically using graphical models (e.g. Bayesian networks). However, inferring the network structure offers a serious challenge in microarray analysis where the sample size is small compared to the number of considered genes. This renders many standard algorithms for graphical models inapplicable, and inferring genetic networks an 'ill-posed' inverse problem. METHODS: We introduce a novel framework for small-sample inference of graphical models from gene expression data. Specifically, we focus on the so-called graphical Gaussian models (GGMs) that are now frequently used to describe gene association networks and to detect conditionally dependent genes. Our new approach is based on (1) improved (regularized) small-sample point estimates of partial correlation, (2) an exact test of edge inclusion with adaptive estimation of the degree of freedom and (3) a heuristic network search based on false discovery rate multiple testing. Steps (2) and (3) correspond to an empirical Bayes estimate of the network topology. RESULTS: Using computer simulations, we investigate the sensitivity (power) and specificity (true negative rate) of the proposed framework to estimate GGMs from microarray data. This shows that it is possible to recover the true network topology with high accuracy even for small-sample datasets. Subsequently, we analyze gene expression data from a breast cancer tumor study and illustrate our approach by inferring a corresponding large-scale gene association network for 3883 genes.

Algorithms↗

Study of the role of functional variants of SLC22A4, RUNX1 and SUMO4 in systemic lupus erythematosus.

BACKGROUND: Functional polymorphisms of the solute carrier family 22, member 4 (SLC22A4), runt related transcription factor 1 (RUNX1) and small ubiquitin-like modifier 4 (SUMO4) genes have been shown to be associated with several autoimmune diseases. OBJECTIVE: To test the possible role of these variants in susceptibility to or severity of systemic lupus erythematosus (SLE), on the basis that common genetic bases are shared by autoimmune disorders. METHODS: 597 SLE patients and 987 healthy controls of white Spanish origin were studied. Two additional cohorts of 228 SLE patients from Sweden and 122 SLE patients from Colombia were included. A case-control association study was carried out with six single nucleotide polymorphisms (SNP) spanning the SLC22A4 gene, one SNP in RUNX1 gene, and one additional SNP in SUM04 gene. RESULTS: No significant differences were observed between SLE patients and healthy controls when comparing the distribution of the genotypes or alleles of any of the SLC22A4, RUNX1, or SUMO4 polymorphisms tested. Significant differences were found in the distribution of the SUMO4 genotypes and alleles among SLE patients with and without nephritis, but after multiple testing correction, the significance of the association was lost. The association of SUMO4 with nephritis could not be verified in two independent SLE cohorts from Sweden and Colombia. CONCLUSIONS: These results suggest that the SLC22A4, RUNX1, and SUMO4 polymorphisms analysed do not play a role in the susceptibility to or severity of SLE.

Adult↗

HIV infection in the Women's Jail, Orange County, California, 1985 through 1991.

The incidence and prevalence of human immunodeficiency virus (HIV) infection among injection drug users, prostitutes, and other women seeking confidential testing in the Orange County Women's Jail were assessed from 1985 to 1991. A total of 4616 voluntary tests were completed on 3051 women, and 865 women were tested repeatedly. Eighty-two women tested positively, a ratio of 1.8 positives per 100 tests or 2.7% of all persons tested. Cumulative HIV prevalence increased from 2.5% to 2.7% between 1985 and 1991, increased by age, and showed racial differences. Of women with multiple tests, 29 seroconverted. Incidence declined from 5.7 to 1.4 cases per 100 person-years of observation between 1985 and 1991. The overall rate of seroconversion was 1.6 per 100 person-years of observation.

Adolescent↗

Minimum muscular fitness in school children.

Two hundred and fifty school boys from Lonavla were randomly selected and tested with Kraus-Weber tests for their minimum muscular fitness. The results revealed that 20.8 per cent boys failed in the tests. Multiple failures were 4.8 percent while flexibility failures alone were 11.6 percent. Boys at the age of 15 years failed more in flexibility test. Boys at the age of 10 years had the maximum percentage of failures to the extent of 38.5. Inclusion of Yogasanas in the programme of physical activities of the school is suggested for the improvement of the status of the failures.

Adolescent↗

The Textarin/Ecarin ratio: a confirmatory test for lupus anticoagulants.

Lupus anticoagulants (LA) are immunoglobulins (IgG, IgM, IgA or a mixture) which interfere with in vitro phospholipid (PL) dependent tests of coagulation (e.g. APTT, KCT, dilute Russell Viper Venom Time). LA are heterogeneous; consequently, the laboratory diagnosis is difficult and relies on multiple tests. We have developed a sensitive and relatively specific confirmatory test system based on fractions of two snake venoms. Textarin, a protein fraction of Pseudonaja textilis venom (Australian Eastern brown snake), activates prothrombin in the presence of PL, factor V and calcium ions. Ecarin, a protein fraction of Echis carinatus venom, will activate prothrombin in the absence of any cofactors. The activation of prothrombin by Textarin yields thrombin while Ecarin yields meizothrombin. In the presence of LA, the Textarin time is prolonged and the Ecarin time is unaffected. The test results are reported as a ratio of Textarin/Ecarin times (abnormal greater than 1.3). We have evaluated this test system in the following patient populations: LA positive, therapeutically heparinized, stable oral anticoagulated, liver disease, routine preoperative, anticardiolipin antibody positive LA negative, hemophilia A, various other hereditary factor deficiencies or dysfunctional proteins, and specific inhibitors of factor V and factor VIII. The LA positive patients represented a mixed population of autoimmune disease, drug-induced and post-infectious states. Our findings indicate the sensitivity of the Textarin/Ecarin system in the confirmation of LA. In order to use the test system most effectively, it is recommended to incorporate polybrene with Textarin when evaluating heparinized samples. Factor V deficiency and specific inhibitors of factor V yielded, in some instances, false positive results.

Administration, Oral↗

Test-ordering by multiple physicians increases unnecessary laboratory examinations.

In modern teaching hospitals, patients typically receive direct care from a succession of different physicians, each of whom may order diagnostic tests on the same patient. We examined the association of test-ordering by multiple physicians with unnecessary duplication of 20-test chemistry profiles in 198 consecutively admitted patients. In a multivariate regression model, the number of duplicate chemistry profiles ordered for a patient was significantly correlated with the number of physicians ordering profiles after controlling for the overall intensity of profile testing. In a case-control analysis comparing duplicate with nonduplicate profiles, redundant tests were significantly more likely to have been ordered by a new physician who had not ordered a patient's previous profile than by the same physician who had ordered the previous chemistry panel. We conclude that test ordering by multiple physicians, the prevalent pattern in almost all teaching hospitals, predisposes to unnecessary laboratory examinations.

Clinical Laboratory Techniques↗

Minimizing errors during in vitro testing of multisegmental spine specimens: considerations for component selection and kinematic measurement.

Apparatus-induced artifacts may invalidate standard spine testing protocols. Kinematic measurements may be compromised by the configuration of motion capture equipment. This study has determined: (1) the influence of machine design (component friction) on in vitro spinal kinetics; (2) the sensitivity of kinematic measurements to variations in the placement of motion capture markers. A spinal loading simulator has been developed to dynamically apply pure bending moments (three axes) with or without a simultaneous compressive preload. Two linear slider types with different friction coefficients, one with caged ball bearings and one with high-precision roller bearings on rails, were mounted and specimen response compared in sequential tests. Three different optoelectronic marker cluster configurations were mounted on the specimen and motion data was captured simultaneously from all clusters during testing. A polymer tube with a uniform bending stiffness approximately equivalent to a polysegmental lumbar spine specimen was selected to allow reproducible behavior over multiple tests. The selection of sliders for linear degrees of freedom had a marked influence on parasitic shear forces. Higher shear forces were recorded with the caged-bearing design than with the high-precision rollers and consequently a higher moment was required to achieve a given rotation. Kinematic accuracy varied with each marker configuration, but in general higher accuracy was achieved with larger marker spacings and situations where markers moved predominantly parallel to the camera's imaging plane. Relatively common alternatives in the mechanical components used in an apparatus for in vitro spine testing can have a significant influence on the measured kinematic and kinetics. Low-magnitude parasitic shear forces due to friction in sliders induces a linearly increasing moment along the length of the specimen, precluding the ideal of pure moment application. This effect is compounded in polysegmental specimens. Kinematic measurements are highly sensitive to marker design and placement, despite equivalent absolute precision of individual marker measurements, however marker configurations can be designed to minimize errors related to spatial distribution and system bias.

Artifacts↗

Measurement invariance in longitudinal clinical research assuming change from development and intervention.

Valid and reliable measures of psychological and behavioral constructs are critical to clinical research, particularly longitudinal treatment research, in which multiple groups are compared over time for possible changes within and between constructs as a result of intervention or development. Structural equation modeling (SEM) analysis is a class of statistical procedures that can be used to test multiple hypotheses about these relationships simultaneously while controlling for measurement error. The procedures have been applied primarily to testing between-construct relationships in nonexperimental studies, with relatively little emphasis on establishing whether measures are sufficiently invariant across groups and across time to permit these tests. This article uses an empirical example of a longitudinal experimental prevention study with two groups to illustrate the use of SEM, first, to systematically test measurement invariance across groups at each wave of measurement, and second, after establishing measurement invariance, to test structural invariance longitudinally.

Adolescent↗

A meta-analytic test of intergroup contact theory.

The present article presents a meta-analytic test of intergroup contact theory. With 713 independent samples from 515 studies, the meta-analysis finds that intergroup contact typically reduces intergroup prejudice. Multiple tests indicate that this finding appears not to result from either participant selection or publication biases, and the more rigorous studies yield larger mean effects. These contact effects typically generalize to the entire outgroup, and they emerge across a broad range of outgroup targets and contact settings. Similar patterns also emerge for samples with racial or ethnic targets and samples with other targets. This result suggests that contact theory, devised originally for racial and ethnic encounters, can be extended to other groups. A global indicator of Allport's optimal contact conditions demonstrates that contact under these conditions typically leads to even greater reduction in prejudice. Closer examination demonstrates that these conditions are best conceptualized as an interrelated bundle rather than as independent factors. Further, the meta-analytic findings indicate that these conditions are not essential for prejudice reduction. Hence, future work should focus on negative factors that prevent intergroup contact from diminishing prejudice as well as the development of a more comprehensive theory of intergroup contact.

Ethnicity↗

Test review: Delis-Kaplan executive function system.

The Delis-Kaplan Executive Function System (D-KEFS; Delis, Kaplan, & Kramer, 2001) represents the first set of executive tests co-normed on a large and representative national sample designed exclusively for the assessment of executive functions including flexibility of thinking, inhibition, problem solving, planning, impulse control, concept formation, abstract thinking, and creativity. The D-KEFS, composed of nine stand alone tests that can be individually or group administered, provides a standardized assessment of executive functions in children and adults between the ages of 8 and 89. Utilizing a "cognitive-process approach," the D-KEFS tests allow examiners to systematically generate and evaluate relevant clinical hypotheses on executive functioning of a given examinee by comparing and contrasting performance on multiple testing conditions and using contrast measure scores and error analyses. A review of the administration, scoring and interpretation, test construction, standardization, and technical adequacy indicate that the D-KEFS holds much promise not only as a clinical instrument, but also as a research tool for increasing knowledge of the frontal-lobe functions.

Humans↗

Cost savings associated with improving appropriate and reducing inappropriate preventive care: cost-consequences analysis.

BACKGROUND: Outreach facilitation has been proven successful in improving the adoption of clinical preventive care guidelines in primary care practice. The net costs and savings of delivering such an intensive intervention need to be understood. We wanted to estimate the proportion of a facilitation intervention cost that is offset and the potential for savings by reducing inappropriate screening tests and increasing appropriate screening tests in 22 intervention primary care practices affecting a population of 90,283 patients. METHODS: A cost-consequences analysis of one successful outreach facilitation intervention was done, taking into account the estimated cost savings to the health system of reducing five inappropriate tests and increasing seven appropriate tests. Multiple data sources were used to calculate costs and cost savings to the government. The cost of the intervention and costs of performing appropriate testing were calculated. Costs averted were calculated by multiplying the number of tests not performed as a result of the intervention. Further downstream cost savings were determined by calculating the direct costs associated with the number of false positive test follow-ups avoided. Treatment costs averted as a result of increasing appropriate testing were similarly calculated. RESULTS: The total cost of the intervention over 12 months was $238,388 and the cost of increasing the delivery of appropriate care was $192,912 for a total cost of $431,300. The savings from reduction in inappropriate testing were $148,568 and from avoiding treatment costs as a result of appropriate testing were $455,464 for a total savings of $604,032. On a yearly basis the net cost saving to the government is $191,733 per year (2003 Can dollars) equating to $3,687 per physician or $63,911 per facilitator, an estimated return on intervention investment and delivery of appropriate preventive care of 40%. CONCLUSION: Outreach facilitation is more expensive but more effective than other attempts to modify primary care practice and all of its costs can be offset through the reduction of inappropriate testing and increasing appropriate testing. Our calculations are based on conservative assumptions. The potential for savings is likely considerably higher.

Adult↗

[The multiple selection vocabulary test (MSVT-B)--an accelerated intelligence test].

A new accelerated intelligence test is presented which is known as the multiple selection vocabulary test B. This is a very objective and reliable test measuring the general level of intelligence. The time of testing will be about three to five minutes in the case of persons that are free of any psychiatric disorders. Test results will be only negligibly influenced by psychic and mental disorders.

Humans↗

Cervicovaginal foetal fibronectin in the prediction of preterm labour in a low-risk population.

The aim of this study was to evaluate the role of cervicovaginal fibronectin as a screening test in a low-risk population. Swabs were taken from the posterior fornix at 28 weeks gestation and foetal fibronectin levels were assayed using enzyme immunoassay. Eighteen patients (7.7%) delivered at less than 37 weeks of gestation, and 6 (2.4%) at less than 34 weeks of gestation. Five patients (2.1%) had a positive fibronectin test result. The sensitivity, specificity, positive predictive value and negative predictive value in the prediction of delivery less than 37 weeks of gestation were 16.7%, 99.1%, 60.0% and 93.4%, respectively. The sensitivity of foetal fibronectin in the prediction of delivery less than 34 weeks of gestation was higher (50%) compared with the prediction of delivery less than 37 weeks of gestation (16.7%). The low sensitivity of this test suggests a limited role for cervicovaginal fibronectin in the low-risk population. Further studies need to be carried out to assess whether multiple testing of cervicovaginal fibronectin at different gestational ages will further increase the sensitivity.

Cervix Uteri↗

Dual HDL/total cholesterol test: a single-tube, homogeneous assay for sequential measurement of HDL cholesterol and total cholesterol.

The quantitation of cholesterol in lipoprotein subfractions is valuable in estimating the risk for coronary artery disease, but requires multiple tests. We describe a relatively simple procedure, referred to as the dual HDL/total cholesterol (DHT) assay, which allows the sequential measurement of HDL cholesterol (HDL-C) and total cholesterol (total-C) in a single tube. HDL-C is first measured using a homogeneous assay that utilizes an anti-apolipoprotein B [apo(B)] antibody, which sterically blocks the enzymatic measurement of cholesterol on the non-HDL subfractions. Next, deoxycholate is added, which disrupts the antibody-apo(B) complex and allows the subsequent enzymatic measurement of the remaining cholesterol in the non-HDL subfractions. The DHT assay has an acceptable analytical performance and yields results similar to standard methods: for HDL-C, y(DHT) = 0.98x + 0.19, r=0.90; for total-C, y(DHT) = 1.11x - 0.09, r=0.99. In summary, the DHT assay is a homogeneous assay for both HDL-C and total-C, and provides a simple and cost-effective method for screening for hyperlipidaemia.

Antibodies↗