OBJECTIVE

The objectives were to give an overview of studies on the validity of the Toronto Alexithymia Scale (TAS-20) and to present data regarding the validity of the TAS-20.

METHODS

The literature on the psychometric properties of the TAS-20 was reviewed and a study was conducted of its psychometric properties in a sample of students and a sample of psychiatric outpatients using a statistical method allowing identification of a stable factor structure.

RESULTS

The review revealed that the majority of studies on the TAS-20 were conducted with nonpatient samples. The factorial validity and reliability of the dimensions 'identifying feelings' (DIF) and 'describing feelings' (DDF) could be replicated in many of these studies. However, in practically all studies the dimension 'externally oriented thinking' (EOT) appears to be unreliable. In addition, the presupposed fantasy aspect of the alexithymia construct is not included in the TAS-20. Although many studies were conducted on the construct validity of the TAS-20, no studies have been published on its criterion validity. Some studies show a different factor structure to exist in patient samples. This was confirmed in our own study in which the dimensions 'identifying feelings' and 'describing feelings' collapsed into one single subscale. As in other studies, the reliability of the dimension 'EOT' was low.

CONCLUSIONS

The TAS-20 has some important shortcomings with respect to validity and reliability. For the assessment of alexithymia in empirical research, it is recommended to use the TAS-20 in combination with other instruments. We do not recommend the TAS-20 to be used in clinical practice.

In cross-national comparisons based on questionnaires, accurate translations are necessary to obtain valid results. Differential item functioning (DIF) analysis can be used to test whether translations of items in multi-item scales are equivalent to the original. In data from 10,815 respondents representing 10 European languages we tested for DIF in the nine translations of the EORTC QLQ-C30 emotional function scale when compared to the original English version. We tested for DIF using two different methods in parallel, a contingency table method and logistic regression. The DIF results obtained with the two methods were similar. We found indications of DIF in seven of the nine translations. At least two of the DIF findings seem to reflect linguistic problems in the translation. 'Imperfect' translations can affect conclusions drawn from cross-national comparisons. Given that translations can never be identical to the original we discuss how findings of DIF can be interpreted and discuss the difference between linguistic DIF and DIF caused by confounding, cross-cultural differences, or DIF in other items in the scale. We conclude that testing for DIF is a useful way to validate questionnaire translations.

In this paper we describe a photon migration approach for modeling fluorescence in an optically thick, turbid medium such as human tissue. In such a medium the intrinsic fluorescence spectrum of the fluorophores Φ can be distorted by the interplay of many factors, including scattering and absorption, excitation and collection geometries, and boundary conditions. The model provides an analytical relationship between the bulk fluorescence spectrum F and the diffuse reflectance spectrum R for arbitrary geometries and boundary conditions. We demonstrate that the distortion can be simply and accurately removed by measuring R from the optically thick medium over the same wavelength range and in the same manner as F. Over a wide range of tissue parameters this relationship may be written as Φα F/R(eff), with R(eff) a corrected form of the measured diffuse reflectance. The validity of this approach is demonstrated in both laboratory experiments on human aortic media and by comparison with Monte Carlo simulations and dif usion theory. Connection with a previous algorithm for extracting intrinsic fluorescence is also discussed.

In bacteria with circular chromosomes, homologous recombination can generate chromosome dimers that cannot be segregated to daughter cells at cell division. Xer site-specific recombination at dif, a 28-bp site located in the replication terminus region of the chromosome, converts dimers to monomers through the sequential action of the XerC and XerD recombinases. Chromosome dimer resolution requires that dif is positioned correctly in the chromosome, and the activity of FtsK, a septum-located protein that coordinates cell division with chromosome segregation. Here, we show that cycles of XerC-mediated strand exchanges form and resolve Holliday junction intermediates back to substrate irrespective of whether conditions support a complete recombination reaction. The C-terminal domain of FtsK is sufficient to activate the exchange of the second pair of strands by XerD, allowing both intra- and intermolecular recombination reactions to go to completion. Proper positioning of dif in the chromosome and of FtsK at the septum is required to sense the multimeric state of newly replicated chromosomes and restrict complete Xer reactions to dimeric chromosomes.

The dimerisation of Raf kinases involves a central cluster within the kinase domain, the dimer interface (DIF). Yet, the importance of the DIF for the signalling potential of wild-type B-Raf (B-Raf(wt)) and its oncogenic counterparts remains unknown. Here, we show that the DIF plays a pivotal role for the activity of B-Raf(wt) and several of its gain-of-function (g-o-f) mutants. In contrast, the B-Raf(V600E), B-Raf(insT) and B-Raf(G469A) oncoproteins are remarkably resistant to mutations in the DIF. However, compared with B-Raf(wt), B-Raf(V600E) displays extended protomer contacts, increased homodimerisation and incorporation into larger protein complexes. In contrast, B-Raf(wt) and Raf-1(wt) mediated signalling triggered by oncogenic Ras as well as the paradoxical activation of Raf-1 by kinase-inactivated B-Raf require an intact DIF. Surprisingly, the B-Raf DIF is not required for dimerisation between Raf-1 and B-Raf, which was inactivated by the D594A mutation, sorafenib or PLX4720. This suggests that paradoxical MEK/ERK activation represents a two-step mechanism consisting of dimerisation and DIF-dependent transactivation. Our data further implicate the Raf DIF as a potential target against Ras-driven Raf-mediated (paradoxical) ERK activation.

BACKGROUND

Demographic differences have been reported in summary measures of physical and mental health based on the SF-12 instrument.

OBJECTIVE

This study examines the extent to which differential item functioning (DIF) contributes to observed subgroup differences in health status. DIF refers to situations in which the psychometric properties of items are not invariant across different groups. The presence of DIF confounds interpretation of subgroup differences.

METHODS

A national sample of 11,626 adult respondents in the 2000 Medical Expenditure Panel Survey who completed a self-administered questionnaire.

METHODS

In addition to the SF-12, we collected data on demographic characteristics (age, gender, education, and race/ethnicity) and whether the person had ever been diagnosed with six chronic medical conditions.

RESULTS

Multiple-indicator multiple-cause latent variable models showed significant differences in physical health by gender, age, and education. Adjusting for DIF reduced but did not eliminate age and education differences. However, for mental health, adjusting for DIF resulted in Black-White differences becoming nonsignificant, and the effect for the oldest age group switched from positive to negative. Race/ethnicity was not associated with physical health status.

CONCLUSIONS

Age group comparisons of mental health may be particularly affected by DIF. Differences in education, as well as age and gender, need to be controlled when making group comparisons. Additional work is needed to understand factors that give rise to demographic differences in reported health status.

OBJECTIVE

To assess the consistency of inclusion and grading of major drug interactions for 50 drugs in four leading international drug interaction compendia. METHODS Four international drug interaction compendia were compared: the drug interactions appendix of the British National Formulary, the interaction supplement in the French drug compendium Vidal, and two US drug interaction compendia, Drug Interaction Facts and the Micromedex (Drug-Reax) program. Major interactions were defined as potentially hazardous in BNF or with the warning 'contraindication' or 'avoid' in Vidal or with the significance grading 1 or 2 in DIF. Major interactions for a list of 50 drugs were searched in all four compendia.

RESULTS

A total of 1264 interactions meeting the inclusion criteria were identified for these 50 drugs. After deletion of 169 duplicates, 1095 interactions were included in the analysis. Of the drug interactions classified as major in any one compendium between 14% and 44% were not listed in the other compendia. The grading systems used for the severity and the quality of the supporting evidence in Micromedex and DIF were inconsistent.

CONCLUSIONS

There is a lack of consistency in the inclusion and grading of drug interactions of major significance for 50 drugs across the four drug compendia examined. This may reflect the lack of standardization of the terminology used to classify drug interactions and the lack of good epidemiological evidence on which to base the assessment of the clinical relevance of drug interactions.

OBJECTIVE

Guidelines have been established for cross-cultural adaptation of outcome measures. However, invariance across cultures must also be demonstrated through analysis of Differential Item Functioning (DIF). This is tested in the context of a Turkish adaptation of the Health Assessment Questionnaire (HAQ).

METHODS

Internal construct validity of the adapted HAQ is assessed by Rasch analysis; reliability, by internal consistency and the intraclass correlation coefficient; external construct validity, by association with impairments and American College of Rheumatology functional stages. Cross-cultural validity is tested through DIF by comparison with data from the UK version of the HAQ.

RESULTS

The adapted version of the HAQ demonstrated good internal construct validity through fit of the data to the Rasch model (mean item fit 0.205; SD 0.998). Reliability was excellent (alpha = 0.97) and external construct validity was confirmed by expected associations. DIF for culture was found in only 1 item.

CONCLUSIONS

Cross-cultural validity was found to be sufficient for use in international studies between the UK and Turkey. Future adaptation of instruments should include analysis of DIF at the field testing stage in the adaptation process.

On starvation, Dictyostelium cells form a terminally differentiated structure, known as the fruiting body, which comprises stalk and spore cells. Their precursors--prestalk and prespore cells--are spatially separated and accessible in a migratory structure known as the slug. This simplicity and manipulability has made Dictyostelium attractive to both experimental and theoretical developmental biologists. However, this outward simplicity conceals a surprising degree of developmental sophistication. Multiple prestalk subtypes are formed and undertake a co-ordinated series of morphogenetic cell movements to generate the fruiting body. This review describes recent advances in understanding the signalling pathways that generate prestalk-cell heterogeneity, focusing on the roles of the prestalk-cell inducer differentiation-inducing factor-1 (DIF-1), the tip inducer cAMP and the transcription factors that mediate their actions; these include signal transducer and activator of transcription (STAT) proteins, basic leucine zipper (bZIP) proteins and a Myb protein of a class previously described only in plants.

In 1986, Chlamydia trachomatis elementary bodies were found by direct immunofluorescence (DIF) in synovial-fluid cell deposits and synovial-membrane biopsy samples from five of eight patients with sexually acquired reactive arthritis (SARA) but in none of eight controls with other types of arthritis. Cells from the original slides (stored at 4 degrees C) have now been examined by a polymerase chain reaction (PCR) that amplifies DNA for the major outer membrane protein of C trachomatis. Chlamydial DNA was found in samples from four DIF-positive patients, one DIF-negative patient, and one DIF-negative control. Overall, there was 80% concordance for DIF and PCR results. This study supports our previous finding of chlamydiae in joints in reactive arthritis.

Sister chromatid exchange (SCE) in Escherichia coli results in the formation of circular dimer chromosomes, which are converted back to monomers by a compensating exchange at the dif resolvase site. Recombination at dif is site specific and can be monitored by utilizing a density label assay that we recently described. To characterize factors affecting SCE frequency, we analyzed dimer resolution at the dif site in a variety of genetic backgrounds and conditions. Recombination at dif was increased by known hyperrecombinogenic mutations such as polA, dut, and uvrD. It was also increased by a fur mutation, which increased oxidative DNA damage. Recombination at dif was eliminated by a recA mutation, reflecting the role of RecA in SCE and virtually all homologous recombination in E. coli. Interestingly, recombination at dif was reduced to approximately half of the wild-type levels by single mutations in either recB or recF, and it was virtually eliminated when both mutations were present. This result demonstrates the importance of both RecBCD and RecF to chromosomal recombination events in wild-type cells.

OBJECTIVE

To develop an instrument to measure subjective quality of vision: the Quality of Vision (QoV) questionnaire.

METHODS

A 30-item instrument was designed with 10 symptoms rated in each of three scales (frequency, severity, and bothersome). The QoV was completed by 900 subjects in groups of spectacle wearers, contact lens wearers, and those having had laser refractive surgery, intraocular refractive surgery, or eye disease and investigated with Rasch analysis and traditional statistics. Validity and reliability were assessed by Rasch fit statistics, principal components analysis (PCA), person separation, differential item functioning (DIF), item targeting, construct validity (correlation with visual acuity, contrast sensitivity, total root mean square [RMS] higher order aberrations [HOA]), and test-retest reliability (two-way random intraclass correlation coefficients [ICC] and 95% repeatability coefficients [R(c)]).

RESULTS

Rasch analysis demonstrated good precision, reliability, and internal consistency for all three scales (mean square infit and outfit within 0.81-1.27; PCA >60% variance explained by the principal component; person separation 2.08, 2.10, and 2.01 respectively; and minimal DIF). Construct validity was indicated by strong correlations with visual acuity, contrast sensitivity and RMS HOA. Test-retest reliability was evidenced by a minimum ICC of 0.867 and a minimum 95% R(c) of 1.55 units.

CONCLUSIONS

The QoV Questionnaire consists of a Rasch-tested, linear-scaled, 30-item instrument on three scales providing a QoV score in terms of symptom frequency, severity, and bothersome. It is suitable for measuring QoV in patients with all types of refractive correction, eye surgery, and eye disease that cause QoV problems.

In Escherichia coli, chromosome dimers are generated by recombination between circular sister chromosomes. Dimers are lethal unless resolved by a system that involves the XerC, XerD and FtsK proteins acting at a site (dif) in the terminus region. Resolution fails if dif is moved from its normal position. To analyse this positional requirement, dif was transplaced to a variety of positions, and deletions and inversions of portions of the dif region were constructed. Resolution occurs only when dif is located at the convergence of multiple, oppositely polarized DNA sequence elements, inferred to lie in the terminus region. These polar elements may position dif at the cell septum and be general features of chromosome organization with a role in nucleoid dynamics.

We have isolated a gene that is very rapidly induced at the transcriptional level by DIF--a low molecular weight, diffusible factor necessary for stalk cell differentiation in Dictyostelium cells developing in vitro. The gene encodes a protein containing an N-terminal signal peptide preceding approximately 70 tandem repeats of a highly conserved 24 amino acid sequence with a high cysteine content. These features suggest it is an extracellular structural protein. During normal development, the gene is maximally expressed in the slug, in which the mRNA is very highly enriched in prestalk over prespore cells. The gene is not detectably expressed until the tipped aggregate stage, several hours later than prespore genes, suggesting that prespore cell differentiation precedes prestalk cell differentiation. The demonstration that DIF induces a gene normally only expressed in the prestalk zone of the slug provides strong evidence that DIF is a Dictyostelium morphogen.

BACKGROUND

Knowledge of the extent to which measurement of adult cognitive functioning differs between Spanish and English language administrations of the Mini-Mental State Examination (MMSE) is critical for inclusive, representative, and valid research of older adults in the United States.

OBJECTIVE

We sought to demonstrate the use of an item response theory (IRT) based structural equation model, that is, the MIMIC model (multiple indicators, multiple causes), to evaluate MMSE responses for evidence of differential item functioning (DIF) attributable to language of administration.

METHODS

We studied participants in a dementia case registry study (n = 1546), 42% of whom were examined with the Spanish language MMSE.

RESULTS

Twelve of 21 items were identified as having significant uniform DIF. The 4 most discrepant included orientation to season, orientation to state, repeat phrase, and follow command. DIF accounted for two-thirds of the observed difference in underlying level of cognitive functioning between Spanish- and English-language administration groups.

CONCLUSIONS

Failing to account for measurement differences may lead to spurious inferences regarding language group differences in level of underlying level of cognitive functioning. The MIMIC model can be used to detect and adjust for such measurement differences in substantive research.

In this article, the authors developed a common strategy for identifying differential item functioning (DIF) items that can be implemented in both the mean and covariance structures method (MACS) and item response theory (IRT). They proposed examining the loadings (discrimination) and the intercept (location) parameters simultaneously using the likelihood ratio test with a free-baseline model and Bonferroni corrected critical p values. They compared the relative efficacy of this approach with alternative implementations for various types and amounts of DIF, sample sizes, numbers of response categories, and amounts of impact (latent mean differences). Results indicated that the proposed strategy was considerably more effective than an alternative approach involving a constrained-baseline model. Both MACS and IRT performed similarly well in the majority of experimental conditions. As expected, MACS performed slightly worse in dichotomous conditions but better than IRT in polytomous cases where sample sizes were small. Also, contrary to popular belief, MACS performed well in conditions where DIF was simulated on item thresholds (item means), and its accuracy was not affected by impact.

In Escherichia coli, the ATP-dependent DNA translocase FtsK transports DNA across the site of cell division and activates recombination by the XerCD recombinases at a specific site on the chromosome, dif, to ensure the last stages of chromosome segregation. DNA transport by FtsK is oriented by 8-base-pair asymmetric sequences ('KOPS'). Here we provide evidence that KOPS promote FtsK loading on DNA and that translocation is oriented at this step.

The aims of this paper are to present findings related to differential item functioning (DIF) in the Patient Reported Outcome Measurement Information System (PROMIS) depression item bank, and to discuss potential threats to the validity of results from studies of DIF. The 32 depression items studied were modified from several widely used instruments. DIF analyses of gender, age and education were performed using a sample of 735 individuals recruited by a survey polling firm. DIF hypotheses were generated by asking content experts to indicate whether or not they expected DIF to be present, and the direction of the DIF with respect to the studied comparison groups. Primary analyses were conducted using the graded item response model (for polytomous, ordered response category data) with likelihood ratio tests of DIF, accompanied by magnitude measures. Sensitivity analyses were performed using other item response models and approaches to DIF detection. Despite some caveats, the items that are recommended for exclusion or for separate calibration were "I felt like crying" and "I had trouble enjoying things that I used to enjoy." The item, "I felt I had no energy," was also flagged as evidencing DIF, and recommended for additional review. On the one hand, false DIF detection (Type 1 error) was controlled to the extent possible by ensuring model fit and purification. On the other hand, power for DIF detection might have been compromised by several factors, including sparse data and small sample sizes. Nonetheless, practical and not just statistical significance should be considered. In this case the overall magnitude and impact of DIF was small for the groups studied, although impact was relatively large for some individuals.

The characteristic structure of the mature Dictyostelium culminant is created by the regionalized cellular differentiation and directed movement of prestalk cells. The front prestalk zone of the migratory slug has previously been considered to be a homogeneous tissue. Here we demonstrate, however, the existence of multiple classes of prestalk cells located in different parts or the slug anterior. The pDd56 and pDd63 genes encoding closely related extracellular matrix proteins are dependent for their expression upon DIF-1, the specific stalk-cell inducer. We have fused the promoters of the two genes to a modified chloramphenicol acetyltransferase (cat) gene to produce immunologically detectable proteins which localize to the cell nucleus. These two markers define three distinct kinds of 'prestalk' cells. One class, which we term 'prestalk A' cells, expressed the pDd63 gene. 'Prestalk B' cells express pDd56 and may also express the pDd63 gene. A third class, which we term 'prestalk 0' cells, expresses neither marker.

The signalling molecule DIF-1 is required for normal cell fate choice and patterning in Dictyostelium. To understand how these developmental processes are regulated will require knowledge of how cells receive and respond to the DIF-1 signal. Previously, we have described a bZIP transcription factor, DimA, which is required for cells to respond to DIF-1. However, it was unknown whether DimA activity is required to activate the DIF response pathway in certain cells or is a component of the response pathway itself. In this study, we describe the identification of a DimA-related bZIP transcription factor, DimB. Rapid changes in the subcellular localisation of both DimA and DimB in response to DIF-1 suggest that they are directly downstream of the DIF-1 signal. Genetic and biochemical interactions between DimA and DimB provides evidence that their ability to regulate diverse targets in response to DIF-1 is partly due to their ability to form homo- and heterodimeric complexes. DimA and DimB are therefore direct regulators of cellular responses to DIF-1.

Drosophila protects itself from infection by microbial organisms by means of its pivotal defense, the so-called innate immunity system. This is its sole defense as it lacks an adaptive immunity system such as is found in mammals. The strong conservation of innate immunity systems in organisms from Drosophila to mammals, and the ease with which Drosophila can be manipulated genetically, makes this fly a good model system for investigating the mechanisms of virulence of a number of medically important pathogens. Potentially damaging endogenous and/or exogenous challenges sensed by specific receptors initiate signals via the Toll and/or Imd signaling pathways. These in turn activate the transcription factors Dorsal, Dorsal-related immune factor (Dif) and Relish, culminating in transcription of genes involved in the production of antimicrobial peptides, melanization, phagocytosis, and the cytoskeletal rearrangement required for appropriate responses. Clarifying the regulatory interactions between the various pathways involved is very important for understanding the specificity and termination mechanism of the immune response.

Unlike most bacteria, Vibrio cholerae harbors two distinct, nonhomologous circular chromosomes (chromosome I and II). Many features of chromosome II are plasmid-like, which raised questions concerning its chromosomal nature. Plasmid replication and segregation are generally not coordinated with the bacterial cell cycle, further calling into question the mechanisms ensuring the synchronous management of chromosome I and II. Maintenance of circular replicons requires the resolution of dimers created by homologous recombination events. In Escherichia coli, chromosome dimers are resolved by the addition of a crossover at a specific site, <em>dif</em>, by two tyrosine recombinases, XerC and XerD. The process is coordinated with cell division through the activity of a DNA translocase, FtsK. Many E. coli plasmids also use XerCD for dimer resolution. However, the process is FtsK-independent. The two chromosomes of the V. cholerae N16961 strain carry divergent dimer resolution sites, <em>dif</em>1 and <em>dif</em>2. Here, we show that V. cholerae FtsK controls the addition of a crossover at <em>dif</em>1 and <em>dif</em>2 by a common pair of Xer recombinases. In addition, we show that specific DNA motifs dictate its orientation of translocation, the distribution of these motifs on chromosome I and chromosome II supporting the idea that FtsK translocation serves to bring together the resolution sites carried by a dimer at the time of cell division. Taken together, these results suggest that the same FtsK-dependent mechanism coordinates dimer resolution with cell division for each of the two V. cholerae chromosomes. Chromosome II dimer resolution thus stands as a bona fide chromosomal process.

Cyclic di-(3′:5′)-guanosine monophosphate (c-di-GMP) is a major prokaryote signalling intermediate that is synthesized by diguanylate cyclases and triggers sessility and biofilm formation. We detected the first eukaryote diguanylate cyclases in all major groups of Dictyostelia. On food depletion, Dictyostelium discoideum amoebas collect into aggregates, which first transform into migrating slugs and then into sessile fruiting structures. These structures consist of a spherical spore mass that is supported by a column of stalk cells and a basal disk. A polyketide, DIF-1, which induces stalk-like cells in vitro, was isolated earlier. However, its role in vivo proved recently to be restricted to basal disk formation. Here we show that the Dictyostelium diguanylate cyclase, DgcA, produces c-di-GMP as the morphogen responsible for stalk cell differentiation. Dictyostelium discoideum DgcA synthesized c-di-GMP in a GTP-dependent manner and was expressed at the slug tip, which is the site of stalk cell differentiation. Disruption of the DgcA gene blocked the transition from slug migration to fructification and the expression of stalk genes. Fructification and stalk formation were restored by exposing DgcA-null slugs to wild-type secretion products or to c-di-GMP. Moreover, c-di-GMP, but not cyclic di-(3′:5′)-adenosine monophosphate, induced stalk gene expression in dilute cell monolayers. Apart from identifying the long-elusive stalk-inducing morphogen, our work also identifies a role for c-di-GMP in eukaryotes.

The developmental bacterium Myxococcus xanthus utilizes gliding motility to aggregate during the formation of multicellular fruiting bodies. The social (S) component of M. xanthus gliding motility requires at least two extracellular surface structures, type IV pili (Tfp) and the fibril polysaccharide or exopolysaccharide (EPS). Retraction of Tfp is proposed to power S motility and EPS from neighbouring cells is suggested to provide an anchor and trigger for Tfp retraction. The production of EPS in M. xanthus is regulated in part by the Dif chemosensory pathway; however, the input signal for the Dif pathway in EPS regulation remains to be uncovered. Using a genetic approach combined with quantitative and qualitative analysis, we demonstrate here that Tfp function upstream of the Dif proteins in regulating EPS production. The requirement of Tfp for the production of EPS was verified using various classes of Tfp mutants. Construction and examination of double and triple mutants indicated that mutations in dif are epistatic to those in pil. Furthermore, extracellular complementation between various Tfp and dif mutants suggests that Tfp, instead of being signals, may constitute the sensor or part of the sensor responsible for mediating signal input into the Dif pathway. We propose that S motility involves a regulatory loop in which EPS triggers Tfp retraction and Tfp provide proximity signals to the Dif pathway to modulate EPS production.