OBJECTIVE

The KIDSCREEN questionnaire for health-related quality of life (HRQOL) assessments in children and adolescents was simultaneously developed across 13 European countries, and it was subsequently translated and culturally adapted to over 30 different languages across the world. The aim of this study was to evaluate the measurement equivalence of the KIDSCREEN-27 across Serbian and Iranian children and adolescents.

METHODS

The items in the KIDSCREEN-27 were analyzed for differential item functioning (DIF) across Iranian and Serbian populations using ordinal logistic regression with three different criteria. The sample included 330 Iranian and 329 Serbian children and adolescents and 330 and 314 of their parents, respectively.

RESULTS

Across the two samples, DIF was detected in 16 (59 %) of 27 items in the child self-reports and in 20 (74 %) of 27 items in the parent/proxy report. However, using alternative criteria based on magnitude detected for DIF, only three items in the parent/proxy report showed significant DIF.

CONCLUSIONS

Our study provided more evidence that the KIDSCREEN-27 possesses DIF items across different cultures, but their impact is probably small, and the questionnaire could be used for cross-cultural HRQOL comparisons.

During Dictyostelium development, the differentiation inducing factor (DIF) triggers expression of the prestalk gene ecmB and induces stalk cell differentiation, a form of programmed cell death. The effects of DIF are mediated by a sustained increase in cytosolic Ca2+ levels. The Ca2+ ATPase inhibitor BHQ causes a similar rise in Ca2+ levels and also induces prestalk gene expression. We show here that Ca2+ is a specific intermediate for prestalk gene induction, since BHQ represses transcription of the cAMP-inducible aggregative gene PDE, the postaggregative gene CP2, and the prespore gene D19. The prestalk gene ecmA is also induced by DIF, but induction appears to occur in two steps, which occur within 1 h and after 2 h, respectively. The slow step shows the same kinetics as ecmB induction and similar to ecmB induction, this step is BHQ inducible and requires an initial round of protein synthesis. The fast step does not require protein synthesis and cannot be induced by BHQ. This indicates that in addition to the slow Ca2+-mediated pathway, there is probably a second fast Ca2+-independent signal transduction pathway for DIF.

OBJECTIVE

Differential item functioning (DIF) assesses the consistency of items on a metric across clinical samples in relation to the attribute being measured. We hypothesized that in older adults with persistent pain, items of the Geriatric Depression Scale (GDS) would evidence DIF based on presence or intensity of pain.

METHODS

Unidimensionality was determined by factor and item analyses. DIF was tested using Rasch Modeling. We then evaluated the psychometric properties of a revised GDS (GDS-PAIN), comprised of items that did not evidence DIF. PATIENT AND SETTINGS: A total of 677 community dwelling older adults (age 65-91) participating in observational or treatment studies of low back or knee pain who endorsed at least moderate pain for at least 3 months. A total of 201 pain-free controls were included in the analysis.

RESULTS

Ten of the 30 items displayed significant DIF. These items were: 1) dropping activities and interests; 2) bothered by persistent thoughts; 3) often get fidgety and restless; 4) prefer to stay home; 5) do not feel full of energy; 6) do not enjoy getting up in the morning; 7) mind is not as clear as it was, 8) feel life is empty; 9) feel more problems with memory; and 10) do not find life very exciting. The modified GDS-PAIN scale did not adversely affect the psychometric properties of the scale.

CONCLUSIONS

The performance of the GDS is affected by pain. When unstable items are removed, the revised GDS (GDS-PAIN) appears to be psychometrically stable and maintains both internal consistency and similar correlation values with a measure of pain as the original scale.

Myxococcus xanthus is a Gram-negative gliding bacterium that aggregates and develops into multicellular fruiting bodies in response to starvation. Two chemosensory systems (frz and dif), both of which are homologous to known chemotaxis proteins, were previously identified through characterization of various developmental mutants. This study aims to examine the interaction between these two systems since both of them are required for fruiting body formation of M. xanthus. Through detailed phenotypic analyses of frz and dif double mutants, we found that both frz and dif are involved in cellular reversal and social motility; however, the frz genes are epistatic in controlling cellular reversal, whereas the dif genes are epistatic in controlling social motility. The study suggests that the integration of these two chemotaxis systems may play a central role in controlling the complicated social behaviors of M. xanthus.

The stalk cell differentiation inducing factor (DIF) has the properties required of a morphogen responsible for pattern regulation during the pseudoplasmodial stage of Dictyostelium development. It induces prestalk cell formation and inhibits prespore cell formation, but there is as yet no strong evidence for a morphogenetic gradient of DIF. We have measured DIF accumulation by monolayers of isolated prestalk and prespore cells in an attempt to provide evidence for such a gradient. DIF is accumulated in the largest quantities by a subpopulation of prestalk cells that specifically express the DIF-inducible genes pDd56 and pDd26. Since it has been shown recently that cells that express pDd56 are localized in the central core of the prestalk cell region of the pseudoplasmodia, our current results suggest a morphogenetic gradient generated by this region.

OBJECTIVE

In two studies, we tested two hypotheses about the relationship of alexithymia to dreaming; that dreams of alexithymic people are barren and rarely recalled, and that the dreams are unregulated and nightmarish.

METHODS

Study 1 was a retrospective survey of dreaming among several hundred young adults, and Study 2 was a 1-week, prospective diary study of 153 young adults in which recall, content, and length of dreams were assessed.

RESULTS

Across both studies, the externally oriented thinking (EOT) facet of the Toronto Alexithymia Scale (TAS)-20 correlated with different dream characteristics than the difficulty identifying feelings (DIF) and difficulty describing feelings (DDF) facets, even after statistically controlling for the other facets. Greater EOT was related to an increased frequency of nights without dream recall, having shorter dreams, having dreams rated as boring and lacking vividness, and not believing in the importance of dreams. In contrast, greater DIF or DDF was related to an increased frequency of nights with disturbing dreams, and having dreams rated as bizarre and aggressive.

CONCLUSIONS

We find support for both hypotheses, but different facets of the multidimensional alexithymia construct account for the two types of dream reports.

We previously reported that differentiation-inducing factor-1 (DIF-1), a morphogen in Dictyostelium discoideum, inhibits the proliferation of human cancer cell lines by inducing β-catenin degradation and suppressing the Wnt/β-catenin signaling pathway. To determine whether β-catenin degradation is essential for the effect of DIF-1, we examined the effect of DIF-1 on human colon cancer cell lines (HCT-116, SW-620 and DLD-1), in which the Wnt/β-catenin signaling pathway is constitutively active. DIF-1 strongly inhibited cell proliferation and arrested the cell cycle in the G(0)/G(1) phase via the suppression of cyclin D1 expression at mRNA and protein levels without reducing β-catenin protein. TCF-dependent transcriptional activity and cyclin D1 promoter activity were revealed to be inhibited via suppression of transcription factor 7-like 2 (TCF7L2) expression. Luciferase reporter assays and EMSAs using the TCF7L2 promoter fragments indicated that the binding site for the transcription factor early growth response-1 (Egr-1), which is located in the -609 to -601 bp region relative to the start codon in the TCF7L2 promoter, was involved in DIF-1 activity. Moreover, RNAi-mediated depletion of endogenous TCF7L2 resulted in reduced cyclin D1 promoter activity and protein expression, and the overexpression of TCF7L2 overrode the inhibition of the TCF-dependent transcriptional activity and cyclin D1 promoter activity induced by DIF-1. Therefore, DIF-1 seemed to inhibit the Wnt/β-catenin signaling pathway by suppressing TCF7L2 expression via reduced Egr-1-dependent transcriptional activity in these colon cancer cell lines. Our results provide a novel insight into the mechanisms by which DIF-1 inhibits the Wnt/β-catenin signaling pathway.

The objective of this study was development and psychometric testing of an adaptive, International Classification of Functioning, Disability, and Health (ICF)-oriented questionnaire to be processed by the rehabilitation physician that aids in assessing mobility, self-care, and domestic life (Moses-Physician). The intent is to develop a physician version, analogous in content to the existing patient questionnaire 'Moses-Patient'. The 58 items of Moses-Patient were converted to an external assessment format without altering the content. The data were compiled for 549 patients with musculoskeletal diseases, 212 patients with cardiac diseases, and 259 neurology rehabilitation patients. Analyses were carried out on the basis of the one-parameter item response theory (Rasch model). Effect sizes and the reliable change index were calculated to test responsiveness. Differential item functioning (DIF) was tested using DIF contrasts, equivalent to Mantel-Haenszel DIF sizes. After the item response theory analysis, 47 of 58 items remained, distributed over 12 scales. The scales are more homogeneous in content than in the patient version because of the omission of 11 items and thus do not cover the ICF categories as broadly. Model fit indices (infit and outfit mean square statistics) were in an acceptable range for all items. Cronbach's alpha was between 0.73 and 0.95. Moreover, there is clear evidence of unidimensionality and sensitivity to change of the scales of Moses-Physician. The item parameters of Moses-Physician are invariant with respect to sex and age for all scales. However, there are clear differences regarding disease groups. The Moses-Physician questionnaire is an adaptive, Rasch-scaled assessment instrument that, to a great extent, covers the contents of the ICF chapters 'mobility', 'self-care', and 'domestic life'.

Bullous pemphigoid (BP) is an acquired bullous disorder that predominantly affects the elderly. It is rare in children but when it occurs, there is considerable clinical and histologic overlap with other acquired or congenital blistering disorders. A definitive diagnosis of childhood BP requires direct immunofluorescence and, in some cases, characterization of the target antigen. Three cases of childhood BP are presented, with their histologic and immunofluorescence findings. The first was a 5-month-old male infant who presented with erythema and bullae of the palms and soles and was found to have linear deposition of IgG and C3 along the dermoepidermal junction on direct immunofluorescence (DIF). Histopathologic examination revealed a subepidermal blister containing eosinophils. Type IV collagen was demonstrated along the floor of the blister cavity by a direct immunoperoxidase technique. The second case was an 8-month-old female infant who presented with a blistering eruption of her palms and soles that then became widespread. Direct immunofluorescence showed linear IgG and C3 at the dermoepidermal junction, with laminin deposition at the base of the blister. The third case was a 7-year-old female with bullae and erosions on the vulva and vaginal mucosa. A subepidermal blister was seen on microscopic examination whereas immunofluorescence demonstrated linear IgG and C3 deposition at the basement membrane zone (BMZ). A literature review uncovered 50 cases of childhood BP confirmed by direct or indirect immunofluorescence, or both, and often with evidence of autoantibodies against either the 180 kD or the 230 kD human bullous pemphigoid antigens (BP180 or BP230). This review was used to delineate characteristics of childhood BP, including the newly proposed subtypes: infantile BP and childhood localized vulval BP. Infantile BP presents within the first year of life and is characterized by BP-like lesions on erythematous or normal acral skin. Localized vulval BP is a self-limited, nonscarring BP-like process that involves only the vulva. Both subtypes are normally self-limited and respond well to either topical or systemic steroids, if treatment is initiated before the disease becomes widespread.

Rasch analysis is now used widely to assess the measurement properties of health status questionnaires. This study tested the stability of the AQ20 - a dichotomous-response measure of health status in asthma, using parameters estimated by a Rasch model. One hundred forty-four asthma patients completed the AQ20 on five occasions over 3 months. At visit 1, two items showed significant misfit and were deleted. At each visit, the overall mean item-person and item-trait interaction statistics from the remaining 18 items (AQ18) were very similar. The repeatability of the item calibrations was excellent (intraclass correlation coefficient 0.95), despite the patients' health having changed (repeated-measures ANOVA: FEV(1) and AQ18 score p<0.0001). Tests of differential item functioning (DIF) over time showed that one item increased in severity. This item refers to ability to garden, and changes in response patterns could be related to seasonal changes over the study period. We conclude that this study has highlighted the usefulness of multiple repeat assessments which allow items to be tested for DIF over time. This is important as inclusion of 'time-dependent' items in a questionnaire may reduce the reliability of the instrument.

In order to determine the infection rate of Chlamydia trachomatis in young males in Austria an epidemiological study was performed on 335 male Austrian soldiers attending the military hospital for a health check-up procedure. Three hundred twenty-nine (98.2%) of the screened males were clinically asymptomatic. Chlamydial diagnosis was established by testing first catch urine (FCU). Urine sediment was tested by an enzyme immunosorbent assay (EIA) and by a direct immunofluorescent test (DIF test). Positive results in both tests were defined "true positives." In 41 (12.2%) of all the 335 soldiers a genital chlamydial infection could be demonstrated by a positive result of the sediment of the FCU in both tests. 93% of the positive results in the DIF test could be confirmed by the EIA whereas only 77% of the positive EIA tests were also positive in the DIF test. Data on genital symptoms and the history of sexually transmitted diseases (STDs) as well as information about sexual relationships were available from all persons included in the study and did not differ between chlamydia positive and negative ones. The study demonstrates a high infection rate with C. trachomatis in mostly asymptomatic young males when using FCU for chlamydial diagnosis. Due to the discrepancy between the EIA and the DIF test, positive results of the sediment of FCU in the EIA test should be confirmed by the DIF test to eliminate false positive cases.

Evaluations of biochemical changes associated with spring-style sled dog racing indicate that differences in cortisol, lactate, and serum glucose levels suggest exercise of moderate duration (but high intensity) has metabolic demands that dif-fer from those for typical endurance sled dog racing. Additionally, hematocrit, albumin, sodium, chloride, and blood urea nitrogen levels decreased in one team of dogs, whereas there were mild increases in sodium, chloride, and blood urea nitrogen in the other team. These opposing biochemical findings suggest physiologic changes associated with differences in hydration status, likely attributed to different dietary and hydration strategies used by the respective kennels.

Background and aims. The aim of this study was to compare shear bond strength (SBS) of metal and ceramic brackets bonded to enamel using acid versus Er:YAG laser etching. Materials and methods. Eighty premolars were divided into 4 groups: AM (acid etching/ metal brackets), AC (acid etching/ ceramic brackets), LM (laser etching/ metal brackets) and LC (laser etching/ ceramic brackets). Enamel condition-ing was done using acid in AC and AM and Er:YAG laser in LC and LM. Brackets were debonded with a Dartec machine and the SBSs were determined. Adhesive remnant index was evaluated under a stereomicroscope. Two additional teeth were conditioned with acid and laser for scanning electron microscopy examination. Comparisons of SBS value were done by ANOVA test. Results. statistical analyses showed that SBSs of acid groups were significantly higher than that of laser groups, but dif-ferences between SBS values of AC/ AM and LC/LM were not significant. SEM examination revealed different etching pattern. Conclusion. Low power Er:YAG laser etching offers clinically acceptable SBS which besides its other superiorities to acid etching can be an appropriate alternative for bonding of ceramic brackets.

A mild cold stress can have positive effects on longevity, aging and resistance to severe stresses in flies (heat, cold, fungal infection), but the causes of these effects remain elusive. In order to know whether these effects could be explained by the DIF transcription factor (a NF-κB-like factor in the Toll innate immunity pathway), the Dif ( 1 ) mutant and its control cn bw strain were subjected to a pretreatment by cold. The DIF factor seems to be involved in the response to fungal infection after a mild cold stress and in the resistance to heat. However, DIF seems to have no role in the increased longevity of non-infected flies and resistance to a severe cold shock, because the cold pretreatment slightly increased longevity in females, mainly in Dif ( 1 ) ones, and resistance to a long cold shock in both sexes of these strains.

Articular cartilage degeneration in the middle carpal joint is a common problem in racing horses. This study evaluated the effect of exercise on the in-vitro synthesis of the large aggregating proteoglycans (aggrecan) and two small proteoglycans, biglycan and decorin, in articular cartilage taken from three weight bearing regions of the third carpal bone of horses which were subjected to moderate or strenuous exercise. Twelve Standardbred horses free from clinical and radiographic disease of the middle carpal joint were subjected to an 8 week moderate exercise program. The horses were then randomly assigned to two groups: group A--continued moderate exercise and group B--strenuous exercise for 17 weeks. Horses were then rested for 16 weeks. Full-depth articular cartilage explants from the dorsal radial facet (DRF), dorsal intermediate facet (DIF) and palmar condyle (PC) of the third carpal bone were collected and cultured. Cartilage proteoglycan content and release into culture media were measured. Newly synthesized proteoglycans were labeled with 35SO4(2-) for 48 h and analyzed by size exclusion and hydrophobic chromatography, sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and autoradiography. Histologic sections of adjacent osteochondral regions were evaluated for evidence of arthritic change. No histologic abnormalities or differences in proteoglycan content were detected in any of the articular cartilage regions examined. There was however, a significant reduction (P < 0.05) in aggrecan synthesis and a concomitant increase in decorin synthesis (P < 0.05) in articular cartilage from the DRF of group B animals. There was no change in biglycan synthesis, aggrecan hydrodynamic size or ability to aggregate in any articular cartilage region. This study has demonstrated that strenuous exercise in horses can lead to a disturbance in the biosynthesis of proteoglycans in articular cartilage regions subjected to high contact stresses (DRF). These metabolic abnormalities, which persisted for 16 weeks after cessation of exercise, could have deleterious effects on the biomechanical properties of the tissue. We suggest that the observed alteration in articular cartilage metabolism in CRF cartilage of strenuously exercised horses could represent a predisposing factor for cartilage degeneration and osteoarthritis at a later stage.

BACKGROUND

No precise studies have been performed on cutaneous leukocytoclastic vasculitis (LV) to establish whether it is better to obtain a skin biopsy from lesional or from perilesional skin for direct immunofluorescence (DIF). There is no agreement on the immunoglobulins most frequently detected and the value of DIF for the classification of cutaneous vasculitis.

METHODS

A prospective study of DIF in lesional and perilesional skin was performed in 50 leukocytoclastic vasculitis patients and 15 nonvasculitis patients.

RESULTS

We detected a higher level of positivity in involved skin than in uninvolved skin for IgG, IgA, IgM, C3 and fibrinogen but not for C1q. In vasculitic patients, IgA was the immunoglobulin most frequently detected in lesional (82%) and perilesional skin (68%), followed by IgM (56 and 34%, respectively) and IgG (20 and 8%, respectively). Only IgA deposits were associated with the diagnosis of vasculitis, with a sensitivity of 82% in lesional and 68% in perilesional skin, and with a specificity of 73 and 66.7%, respectively. The presence of IgA in lesional skin was associated with renal involvement but there was no association with severity. The presence of IgG or IgM, or the absence of IgA in perilesional skin was related to the presence of cryoglobulins. The absence of IgA in lesional and perilesional skin was also related to hepatitis C virus infection.

CONCLUSIONS

DIF findings in involved skin are more closely related to the diagnosis of vasculitis and can give more information about overall renal involvement than findings in uninvolved skin. However, findings in uninvolved skin are more closely related to the pathogenic factors that trigger the development of vasculitis.

Fluoroquinolones like difloxacin (DIF) and sarafloxacin (SARA) are adsorbed in soil and enter the aquatic environment wherein they are subjected to photolytic degradation. To evaluate the fate of DIF and SARA, their photolysis was performed in water under stimulated natural sunlight conditions. DIF primarily degrades to SARA. On prolonged photodegradation, seven photoproducts were elucidated by HR-LC-MS/MS, three of which were entirely novel. The residual anti-bacterial activities of DIF, SARA and their photoproducts were studied against a group of pathogenic strains. DIF and SARA revealed potency against both gram-positive and -negative bacteria. The photoproducts also exhibited varying degrees of efficacies against the tested bacteria. Even without isolating the individual photoproducts, their impact on the aquatic environment could be assessed. Therefore, the present results call for prudence in estimating the fate of these compounds in water and in avoiding emergence of resistance in bacteria caused by the photoproducts of DIF and SARA.

During formation of the Dictyostelium slug extracellular cAMP signals direct the differentiation of prespore cells and DIF, a chlorinated hexaphenone, induces the differentiation of prestalk cells. At culmination the slug transforms into a fruiting body, composed of a stalk supporting a ball of spores. A dominant inhibitor of cAMP-dependent protein kinase (PKA) expressed under the control of a prestalk-specific promoter blocks the differentiation of prestalk cells into stalk cells. Analysis of a gene specifically expressed in stalk cells suggests that PKA acts to remove a repressor that prevents the premature induction of stalk cell differentiation by DIF during slug migration. PKA is also necessary for the morphogenetic movement of prestalk cells at culmination. Expression of the PKA inhibitor under control of a prespore-specific promoter blocks the accumulation of prespore mRNA sequences and prevents terminal spore cell differentiation. Thus PKA is essential for progression along both pathways of terminal differentiation but with different mechanisms of action. On the stalk cell pathway it acts to regulate the action of DIF while on the spore cell pathway PKA itself seems to act as the inducer of spore cell maturation. Ammonia, the extracellular signal which regulates the entry into culmination, acts by controlling the intracellular concentration of cAMP and thus exerts its effects via PKA. The fact that PKA is necessary for both prespore and spore gene expression leads us to postulate the existence of a signalling mechanism which converts the progressive rise in cAMP concentration during development into discrete, PKA-regulated gene activation events.

The purpose of this article is to introduce the methods used and challenges confronted by the authors of this two-part series of articles describing the results of analyses of measurement equivalence of the short form scales from the Patient Reported Outcomes Measurement Information System® (PROMIS®). Qualitative and quantitative approaches used to examine differential item functioning (DIF) are reviewed briefly. Qualitative methods focused on generation of DIF hypotheses. The basic quantitative approaches used all rely on a latent variable model, and examine parameters either derived directly from item response theory (IRT) or from structural equation models (SEM). A key methods focus of these articles is to describe state-of-the art approaches to examination of measurement equivalence in eight domains: physical health, pain, fatigue, sleep, depression, anxiety, cognition, and social function. These articles represent the first time that DIF has been examined systematically in the PROMIS short form measures, particularly among ethnically diverse groups. This is also the first set of analyses to examine the performance of PROMIS short forms in patients with cancer. Latent variable model state-of-the-art methods for examining measurement equivalence are introduced briefly in this paper to orient readers to the approaches adopted in this set of papers. Several methodological challenges underlying (DIF-free) anchor item selection and model assumption violations are presented as a backdrop for the articles in this two-part series on measurement equivalence of PROMIS measures.

Right ventricular (RV) pacing has been linked with lead-induced tricuspid regurgitation (TR), left ventricular (LV) dysfunction, and dyssynchrony, but the effect of pacing on RV function is unclear. We sought to investigate the effect of pacing on RV synchrony, RV function, and TR, and their association with LV function. In this substudy of the PROTECT-PACE (Protection of left ventricular function during right ventricular pacing) study of the effects of RV pacing in patients with preserved ejection fraction, 145 patients (76 RV apex and 69 non-RV apex pacing) had measurable RV parameters. We assessed tricuspid annular plane systolic excursion (TAPSE), time difference between peak TAPSE and peak mitral annular plane systolic excursion (TM-APSE-dif), global LV longitudinal strain, E/e', TR vena contracta, and TR peak gradient. Echocardiography was performed just after implantation and at 2 years. TR parameters significantly worsened after 2 years, but pacing site was not associated with changes in RV and TR parameters. No temporal change in TAPSE and TM-APSE-dif was observed overall, but worsening of TM-APSE-dif was associated with worsening TAPSE. Global longitudinal strain and E/e' both deteriorated over 2 years; these changes were significantly associated. In a multivariate regression, worsening global longitudinal strain and worsening TM-APSE-dif were significantly associated with deterioration of TAPSE. Furthermore, increased E/e' and its deterioration were associated with worsening TR vena contracta and TR peak gradient, respectively. Decreased TAPSE was also associated with deterioration of TR vena contracta. In conclusion, RV pacing appears to worsen TR, an effect which might be caused by elevated LV filling pressure due to LV dysfunction. In this study, RV pacing did not affect RV function during 2-year follow-up, but such an effect might occur if RV dyssynchrony or LV dysfunction occurred after pacing. The effect of RV pacing site on RV and TR mechanics was minor.

BACKGROUND

Xerostomia is a symptom that can be triggered by chronic diseases such as Sjögren's syndrome (SS) and lupus erythematosus (LE). Many authors regard most cases of salivary hypofunction in LE to secondary SS. Others believe that salivary changes in patients with LE might reflect a multisystem presentation of the disease. The present study compared histopathological and direct immunofluorescence (DIF) alterations in salivary glands of patients with xerostomia and diagnosis of LE or SS.

METHODS

Twenty-eight salivary gland biopsies from patients with xerostomia and diagnosed with LE or SS were submitted to histopathological and DIF exams.

RESULTS

From the 28 patients, 16 had SS and 12 had LE. In SS, a moderate to intense sialadenitis was detected, with infiltration and destruction of excretory salivary ducts. In LE, mild/moderate sialadenitis with thickening and hyalinization of the ductal basement membrane was observed. DIF revealed that 50% of SS patients presented intercellular ductal IgA deposits, whereas 58% of LE patients showed deposits of IgG in the ductal basement membrane.

CONCLUSIONS

Alterations in salivary glands of LE patients may be a specific manifestation of the disease (lupus sialadenitis), reflecting its multisystemic presentation, instead of an association of secondary SS and LE.

There is suggestive evidence that a conserved signalling system involving differentiation inducing factor-1 (DIF-1) controls stalk cell differentiation in a variety of slime mould species. In the standard laboratory species, Dictyostelium discoideum, DIF-1 is first inactivated by dechlorination catalysed by DIF-1 dechlorinase, then by several hydroxylation events, so that eventually about 12 metabolites are produced. If DIF-1 is used as a signal molecule in other species, they too must be able to metabolize it. We report here that the essentials of DIF-1 metabolism are conserved in D. mucoroides, the closest relative of D. discoideum. Both the dechlorinase and hydroxylase enzymes were present in D. mucoroides, and living cells of both species produced a similar spectrum of metabolites from [3H]DIF-1. Furthermore, DIF-1 dechlorinase was induced by DIF-1, as in D. discoideum, and this induction was repressed by ammonia and cAMP. DIF-1 dechlorinase could not be detected in cell extracts from D. minutum or Polysphondylium violaceum. However, living cells of both species are able to metabolize DIF-1; P. violaceum seems to produce a small amount of the monodechlorinated compound, DIF-3, but all other metabolites from both species appear to be unique. Thus all investigated species can metabolize DIF-1, but the exact route of metabolism is not highly conserved.

Dictyostelium development is orchestrated by diffusible signals. Progress has been made in understanding how cAMP signaling triggers post-aggregative development and in defining the number of cell types that eventually differentiate. Ammonia is an unusual signal that may act by alkalinizing acidic vesicles. A chlorinated signal, differentiation-inducing factor (DIF), may be universal amongst the slime moulds. The first genes have been cloned using restriction enzyme mediated integration (REMI) insertional mutagenesis; one encodes a novel cytosolic protein essential for activation of adenylyl cyclase.

Fluoroquinolones are amongst the most important antibiotics used in veterinary medicine. On this account the behavior of difloxacin (DIF) and its metabolites was investigated by administering the (14)C-labeled and non-labeled veterinary drug to fattening pigs. The excretion kinetics were determined after daily collection of manure. Sarafloxacin (SAR) was found to be the major metabolite, three further trace metabolites were also recovered, applying high-resolution (HR) mass spectrometric technique. The identification of DIF and SAR was confirmed by comparison with the spectroscopic and chromatographic data of the authentic references. The identification of the three trace metabolites was performed by HR-MS/MS. Only 8.1% of the administered radioactivity remained in the pig after 10 days and DIF accounted for 95.9% of the radioactivity excreted. More than 99% of the labeled compounds were detected and identified in the manure. The mean recoveries for all single electrolytes were 94%. Linearity was established over concentration range 10-10,000 microg/kg manure with a correlation coefficient 0.99. By using in vitro antimicrobial activity tests against a group of standard pathogenic control strains, the results showed that the residual antibiotic concentrations in the manure of pigs are high enough to exhibit antibacterial activity.