Immunity genes are activated in the Drosophila fat body by Rel and GATA transcription factors. Here, we present evidence that an additional regulatory factor, deformed epidermal autoregulatory factor-1 (DEAF-1), also contributes to the immune response and is specifically important for the induction of two genes encoding antimicrobial peptides, Metchnikowin (Mtk) and Drosomycin (Drs). The systematic mutagenesis of a minimal Mtk 5' enhancer identified a sequence motif essential for both a response to LPS preparations in S2 cells and activation in the larval fat body in response to bacterial infection. Using affinity chromatography coupled to multidimensional protein identification technology (MudPIT), we identified DEAF-1 as a candidate regulator. DEAF-1 activates the expression of Mtk and Drs promoter-luciferase fusion genes in S2 cells. SELEX assays and footprinting data indicate that DEAF-1 binds to and activates Mtk and Drs regulatory DNAs via a TTCGGBT motif. The insertion of this motif into the Diptericin (Dpt) regulatory region confers DEAF-1 responsiveness to this normally DEAF-1-independent enhancer. The coexpression of DEAF-1 with Dorsal, Dif, and Relish results in the synergistic activation of transcription. We propose that DEAF-1 is a regulator of Drosophila immunity.

Aim: There is a critical need for effective therapies that are immediately available to control the spread of COVID-19 disease. Material & methods: Gamunex^®-C and Flebogamma^® DIF (Grifols) intravenous immunoglobulin (IVIG) products were tested using ELISA techniques for antibodies against several antigens of human common betacoronaviruses that may crossreact with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. Results: Both IVIGs showed consistent reactivity to components of the tested viruses. Positive crossreactivity was seen in SARS-CoV, middle east respiratory syndrome-CoV and SARS-CoV-2. For SARS-CoV-2, positive reactivity was observed at IVIG concentrations ranging from 100 μg/ml with Gamunex-C to 1 mg/ml with Flebogamma 5% DIF. Conclusion: Gamunex-C and Flebogamma DIF contain antibodies reacting against SARS-CoV-2 antigens. Studies to confirm the utility of IVIG preparations for COVID-19 management may be warranted.

OBJECTIVE

To examine the psychometrics of the Affiliate Stigma Scale using rigorous psychometric analysis: classical test theory (CTT) (traditional) and Rasch analysis (modern). Differential item functioning (DIF) items were also tested using Rasch analysis.

METHODS

Caregivers of relatives with mental illness (n = 453; mean age: 53.29 ± 13.50 years) were recruited from southern Taiwan. Each participant filled out four questionnaires: Affiliate Stigma Scale, Rosenberg Self-Esteem Scale, Beck Anxiety Inventory, and one background information sheet.

RESULTS

CTT analyses showed that the Affiliate Stigma Scale had satisfactory internal consistency (α = 0.85-0.94) and concurrent validity (Rosenberg Self-Esteem Scale: r = -0.52 to -0.46; Beck Anxiety Inventory: r = 0.27-0.34). Rasch analyses supported the unidimensionality of three domains in the Affiliate Stigma Scale and indicated four DIF items (affect domain: 1; cognitive domain: 3) across gender.

CONCLUSIONS

Our findings, based on rigorous statistical analysis, verified the psychometrics of the Affiliate Stigma Scale and reported its DIF items. We conclude that the three domains of the Affiliate Stigma Scale can be separately used and are suitable for measuring the affiliate stigma of caregivers of relatives with mental illness.

BACKGROUND

Anti-neutrophil p-ANCA antibodies are directed against antigens in the peripheral cytoplasm of both neutrophilic granulocytes and monocytes. They are detected in several autoimmune disorders and are particularly associated with systemic vasculitis.

METHODS

We report a case of a 54-year-old female presenting with a pruritic rash, including purpura and diffuse erythema. A biopsy with hematoxylin and eosin (H & E) analysis, direct immunofluorescence (DIF), immunohistochemistry (IHC) and enzyme-linked immunosorbent assays for ANCAs were performed. The H & E staining demonstrated leukocytoclastic vasculitis, with focal vascular fibrinoid necrosis. The DIF revealed evidence of vasculitis, the presence of p-ANCAs and neutrophil extracellular traps (NETs). The IHC displayed autoreactivity to myeloperoxidase within the vessels. The IHC aided in ruling out any intrinsic autofluorescence of the vessels.

CONCLUSIONS

By observing the deposition of neutrophil extracellular traps and myeloperoxidase in inflamed skin vessels, biopsy analysis may alert physicians for rapid therapeutic intervention in patients presenting with possible vasculitides.

Dermatitis herpetiformis (DH) is an inflammatory cutaneous disease with a chronic relapsing course, pruritic polymorphic lesions, and typical histopathological and immunopathological findings. According to several evidences, DH is considered the specific cutaneous manifestation of celiac disease, and the most recent guidelines of celiac disease have stated that, in celiac patients with a proven DH, a duodenal biopsy is unnecessary for the diagnosis. In this review, the most recent data about the diagnosis and the management of DH have been reported and discussed. In particular, in patients with clinical and/or histopathological findings suggestive for DH, the finding of granular IgA deposits along the dermal-epidermal junction or at the papillary tips by direct immunofluorescence (DIF) assay, together with positive results for anti-tissue transglutaminase antibody testing, allows the diagnosis. Thereafter, a gluten-free diet should be started in association with drugs, such as dapsone, that are able to control the skin manifestations during the first phases of the diet. In conclusion, although DH is a rare autoimmune disease with specific immunopathological alterations at the skin level, its importance goes beyond the skin itself and may have a big impact on the general health status and the quality of life of the patients.

The Skindex is a well-studied dermatology-specific health-related quality of life (HRQOL) instrument. The objective of this study was to test Skindex-29 using Rasch analysis and, if necessary, to refine it so that it would fit this item response theory based model. The Skindex-29 of 454 Italian dermatological patients was subjected to Rasch analysis to investigate threshold order, differential item functioning (DIF), and item and overall fit to the model. The Skindex-29 did not fit the Rasch model (P<0.001). The 5-point scoring system was re-grouped into three categories and demonstrated logical response order for all but one item. Rasch analyses of a combined emotion and social functioning subscale of Skindex-29 resulted in a 12-item psychosocial subscale. Five of seven items were retained in a symptoms subscale. Both subscales fitted the model (P=0.32 and 0.13, respectively) without significant individual item misfit or DIF (P>0.05). Classical psychometric properties such as response distribution, item-rest correlation, item complexity, and internal consistency of the two subscales of Skindex-17 were at least adequate. The Skindex-17 is a Rasch reduced version of Skindex-29, with two independent scores that can be used in the measurement of HRQOL in dermatological patients.

The objectives of this study were (a) to determine the concurrent validity of the flight time (FT) and double integration of vertical reaction force (DIF) methods in the estimation of vertical jump height with the video method (VID) as reference; (b) to verify the degree of agreement among the 3 methods; (c) to propose regression equations to predict the jump height using the FT and DIF. Twenty healthy male and female nonathlete college students participated in this study. The experiment involved positioning a contact mat (CTM) on the force platform (FP), with a video camera 3 m from the FP and perpendicular to the sagittal plane of the subject being assessed. Each participant performed 15 countermovement jumps with 60-second intervals between the trials. Significant differences were found between the jump height obtained by VID and the results with FT (p ≤ 0.01) and DIF (p ≤ 0.01), showing that the methods are not valid. Additionally, the DIF showed a greater degree of agreement with the reference method than the FT did, and both presented a systematic error. From the linear regression test was determined the prediction equations with a high degree of linearity between the methods VID vs. DIF (R = 0.988) and VID vs. FT (R = 0.979). Therefore, the prediction equations suggested may allow coaches to measure the vertical jump performance of athletes by the FT and DIF, using a CTM or an FP, which represents more practical and viable approaches in the sports field; comparisons can then be made with the results of other athletes evaluated by VID.

Two distinct roles are described for Dorsal, Dif and Relish, the three NF-kappaB/Rel proteins of Drosophila, in the development of the peripheral nervous system. First, these factors regulate transcription of scute during the singling out of sensory organ precursors from clusters of cells expressing the proneural genes achaete and scute. This effect is possibly mediated through binding sites for NF-kappaB/Rel proteins in a regulatory module of the scute gene required for maintenance of scute expression in precursors as well as repression in cells surrounding precursors. Second, genetic evidence suggests that the receptor Toll-8, Relish, Dif and Dorsal, and the caspase Dredd pathway are active over the entire imaginal disc epithelium, but Toll-8 expression is excluded from sensory organ precursors. Relish promotes rapid turnover of transcripts of the target genes scute and asense through an indirect, post-transcriptional mechanism. We propose that this buffering of gene expression levels serves to keep the neuro-epithelium constantly poised for neurogenesis.

We have identified a novel component, Helicase89B, that is required for the inducible antimicrobial response in Drosophila larvae by means of a P-element insertional genetic screen. Helicase89B belongs to the Mot1p/BTAF1 subfamily of SNF2-like ATPases. This subfamily can interact with TATA-binding proteins, but whether the interaction leads to gene activation or repression is being debated. We found that Helicase89B is required for the inducible expression of antimicrobial peptide genes but not for the inducible expression of heat-shock genes. The antimicrobial peptide genes are activated by the Toll and immune deficiency (IMD) signalling pathways. Genetic experiments show that Helicase89B acts downstream of DIF and Relish, the two nuclear factor-kappaB (NF-kappaB)-related transcription factors that mediate Toll- and IMD-stimulated antimicrobial response. Thus, Helicase89B positively regulates gene expression during innate immune response and may act as a link between NF-kappaB-related transcription factors and the basal transcription machinery.

BACKGROUND

Differential item functioning (DIF) exists when test item responses by members of different demographic groups are statistically different when controlling for ability. DIF may indicate item bias. Our objective was to determine whether items from the Italian Mini-mental State Examination (MMSE) had DIF related to educational attainment, age, gender and occupation. We were also interested in exploring the significance of DIF in screening tests.

METHODS

In a two-stage study from Granarolo, Italy, residents over age 61 (n = 495) were evaluated with the Italian MMSE. Those with MMSE scores of 28 or lower were further evaluated for dementia. MMSE results were coded in 10 item bundles. We used ordinal logistic regression to determine whether item bundles had DIF.

RESULTS

Six of the 10 MMSE item bundles had DIF in educational attainment subgroups. Four of these six bundles also had DIF related to age. Items that required literacy were much harder for those with lower educational attainment.

CONCLUSIONS

DIF related to education appeared at as few as 3 years of formal schooling. These findings suggest cautious interpretation of data from studies using the Italian MMSE in populations with heterogeneous educational backgrounds. DIF is especially troublesome for two-stage studies that use screening tests.

Bacteria harbouring circular chromosomes have a Xer site-specific recombination system that resolves chromosome dimers at division. In Escherichia coli, the activity of the XerCD/dif system is controlled and coupled with cell division by the FtsK DNA translocase. Most Xer systems, as XerCD/dif, include two different recombinases. However, some, as the Lactococcus lactis XerS/dif(SL) system, include only one recombinase. We investigated the functional effects of this difference by studying the XerS/dif(SL) system. XerS bound and recombined dif(SL) sites in vitro, both activities displaying asymmetric characteristics. Resolution of chromosome dimers by XerS/dif(SL) required translocation by division septum-borne FtsK. The translocase domain of L. lactis FtsK supported recombination by XerCD/dif, just as E. coli FtsK supports recombination by XerS/dif(SL). Thus, the FtsK-dependent coupling of chromosome segregation with cell division extends to non-rod-shaped bacteria and outside the phylum Proteobacteria. Both the XerCD/dif and XerS/dif(SL) recombination systems require the control activities of the FtsKγ subdomain. However, FtsKγ activates recombination through different mechanisms in these two Xer systems. We show that FtsKγ alone activates XerCD/dif recombination. In contrast, both FtsKγ and the translocation motor are required to activate XerS/dif(SL) recombination. These findings have implications for the mechanisms by which FtsK activates recombination.

OBJECTIVE

Rasch item response theory analysis is essential in evaluating measurement tools in specific disease cohorts. We compared the performance of 4 functional indexes in patients with psoriatic arthritis (PsA) in axial or peripheral disease subgroups.

METHODS

A cross-sectional study was performed in a single center. Functional outcomes assessed by the Health Assessment Questionnaire (HAQ), Bath Ankylosing Spondylitis Functional Index (BASFI), Dougados Functional Index (FI), and the physical functioning scale of the Medical Outcome Study Short-form 36 (SF-36-PF) were analyzed by the Rasch model for item fit, item separation, measurement span, and distribution properties. Patient subgroups with axial or peripheral disease were analyzed for differential item functioning (DIF).

RESULTS

One hundred eight patients with PsA were assessed. The 4 functional indexes were highly correlated with each other and moderately correlated with patients' perception of health and pain scores. Floor effects were less marked in SF-36-PF. The 4 indexes satisfied the unidimensionality assumption of the Rasch model. HAQ and SF-36-PF had better information-weighted fit statistics (INFIT) and outlier-sensitive (OUTFIT) statistics. HAQ had the poorest item separation. SF-36-PF had the highest item separation (6.99), reliability (0.85), and the longest span of item threshold (9.03 logits). Only 1 and 2 items in BASFI and Dougados-FI had DIF in patients with sacroiliitis.

CONCLUSIONS

HAQ, BASFI, Dougados-FI, and SF-36-PF provide unidimensional measures of functional disability in PsA. SF-36-PF was the best in terms of less floor effect, highest item separation, longest span of item threshold, and better distributional properties. BASFI and Dougados-FI behaved similarly in patients with and without sacroiliitis and conferred no superiority in patients with axial disease.

Comparisons were made between the Direct Antiglobulin Rosetting Reaction (DARR) and Direct Immunofluorescence (DIF) in the detection of surface membrane immunoglobulin of human peripheral lymphocytes. The DARR was more sensitive and the results with this testing procedure (as opposed to those with the DIF) were not influenced by various treatments of the lymphocytes before testing, such as incubation in AB serum at +/- 4 degrees, incubation in serum-free medium at 37 degrees or short exposure to acetate buffer at pH 4.0. Again the DARR (as opposed to the DIF) gave essentially the same results whether the red cell-linked antiglobulin was IgG or the F(ab')2 preparation. With mixed rosetting for both T and SmIg+/- lymphocytes, there was only 1% or less null cells and only 5% or less lymphocytes rosetted with both marker red cells.

Genomic clones of the genes coding for the three major spore coat proteins, SP60, SP70, and SP96, were used to measure the accumulation of their respective mRNAs in mutant and wild-type cells allowed to develop under a variety of conditions. These prespore-specific mRNAs were found to be both temporally and quantitatively coordinate under all conditions indicating that they may be subject to identical regulatory processes. Accumulation of the spore coat mRNAs is dependent upon the function of both cAMP receptors and G alpha 2 proteins during the aggregation stage as well as upon concomitant protein synthesis. When cells are dissociated from aggregates at 10 hr of development and rapidly shaken in 0.1 mM EDTA they form clumps but do not accumulate any of the prespore-specific RNAs assayed. However, if either 0.1 mM Ca++ or 20 microM cAMP is added to these cells, the spore coat mRNAs accumulate. Lower concentrations of either Ca++ or cAMP had no effect. These results suggest that expression of the spore coat genes normally involves a Ca+(+)-dependent process, but the Ca++ requirement can be overcome by adding high concentrations of exogenous cAMP. Addition of 50 nM DIF to dissociated cell blocks the accumulation of the spore coat mRNAs even when cAMP or Ca++ is present. The upstream regions of the spore coat genes were compared to those of another gene, D19, that codes for the prespore-specific protein SP29. Short sequences related to CACCCAC were found at about the same position relative to the transcriptional start sites of these coordinately regulated genes.

Dihydropyrimidine dehydrogenase (DPD) is the rate-limiting enzyme in the degradation of pyrimidine bases. DPD is also responsible for the degradation of 5-fluorouracil (5-FU), which is the most frequently prescribed anticancer drug for the treatment of malignancies of the gastrointestinal tract. DPD could influence the antitumor effect and the adverse effects of 5-FU. High intratumoral DPD activity markedly decreases the cytotoxic effect of 5-FU. More than 80% of administered 5-FU is detoxified and excreted as F-beta-alanine in urine. In 5-FU-based chemotherapy, escape from the degradation catalyzed by DPD is important. Recently, the dihydropyrimidine dehydrogenase gene (DPYD) was isolated, and its physical map and exon-intron organization were determined. To date, many DPYD variant alleles associated with a lack of DPD activity have been identified. In 5-FU-based cancer chemotherapy, severe toxicities were observed at higher rates in patients who were heterozygous for a mutant DPYD allele, compared with toxicities in patients who were homozygous for the wild DPYD allele. Furthermore, the adverse effects of 5-FU are often lethal for patients homozygous for the mutant DPYD allele. The apparently high prevalence of the DPYD mutation associated with lack of DPD activity in the normal population warrants genetic screening for the presence of these mutations in cancer patients before the administration of 5-FU. DPD inhibitory fluoropyrimidines (DIFs), including uracil plus tegafur (UFT) and tegafur plus 5-chloro-2,4-dihydroxypyridine plus potassium oxonate, in a molar ratio of 1:0.4:1 (TS-1), have recently been used in clinical settings. DIFs should provide chemotherapy that improves both quality of life and duration of survival.

BACKGROUND

The Drosophila Toll pathway takes part in both establishment of the embryonic dorsoventral axis and induction of the innate immune response in adults. Upon activation by the cytokine Spätzle, Toll interacts with the adaptor proteins DmMyD88 and Tube and the kinase Pelle and triggers degradation of the inhibitor Cactus, thus allowing the nuclear translocation of the transcription factor Dorsal/Dif. weckle (wek) was previously identified as a new dorsal group gene that encodes a putative zinc finger transcription factor. However, its role in the Toll pathway was unknown.

RESULTS

Here, we isolated new wek alleles and demonstrated that cactus is epistatic to wek, which in turn is epistatic to Toll. Consistent with this, Wek localizes to the plasma membrane of embryos, independently of Toll signaling. Wek homodimerizes and associates with Toll. Moreover, Wek binds to and localizes DmMyD88 to the plasma membrane. Thus, Wek acts as an adaptor to assemble/stabilize a Toll/Wek/DmMyD88/Tube complex. Remarkably, unlike the DmMyD88/tube/pelle/cactus gene cassette of the Toll pathway, wek plays a minimal role, if any, in the immune defense against Gram-positive bacteria and fungi.

CONCLUSIONS

We conclude that Wek is an adaptor to link Toll and DmMyD88 and is required for efficient recruitment of DmMyD88 to Toll. Unexpectedly, wek is dispensable for innate immune response, thus revealing differences in the Toll-mediated activation of Dorsal in the embryo and Dif in the fat body of adult flies.

A recessive semi-lethal mutation resulting from the insertion of a P-lacW transposon at the cytological position 23A on the polytene chromosomes of Drosophila melanogaster was found to affect the unfolding and expansion of the wings resulting in a loss of venation and a marked decrease in their size. Lethality was polyphasic with numerous animals dying during early larval development and displaying apparently collapsed tracheal trees. The gene was therefore designated as congested-like tracheae, or colt. The colt mutation resulted from the insertion of a P-lacW transposon within the coding region of a 1.4-kb transcript. Wild-type function was restored by inducing a precise excision of the P-lacW transposon, while a deletion of the colt locus, produced by imprecise excision of the P element, showed a phenotype similar to that of the original P insert. The colt gene consists of a single exon and encodes a protein of 306 amino acids made of three tandem repeats, each characterized by two predicted transmembrane segments and a loop domain. The COLT protein shares extensive homology with proteins in the mitochondrial carrier family and particularly with the DIF-1 protein of Caenorhabditis elegans, which has been shown to be maternally required for embryonic tissue differentiation. Our analysis revealed that zygotic colt function is dispensable for normal embryonic morphogenesis but is required for gas-filling of the tracheal system at hatching time of the embryo and for normal epithelial morphogenesis of the wings.

Recent work has highlighted two main levels of global organization of the Escherichia coli chromosome. Macrodomains are large domains inferred from structural data consisting of loci showing the same intracellular positioning. Replichores, defined by base composition skews, coincide with the replication arms in normal cells. We used chromosome inversions to show that the dif site, which resolves chromosome dimers, only functions when located at the junction of the replichores, whatever their size. This is the first evidence that replichore polarization has a role in chromosome segregation. We also show that disruption of the Ter macrodomain provokes a cell-cycle defect independent from dimer resolution. This confirms the existence of the Ter macrodomain and suggests a role in chromosome dynamics.

OBJECTIVE

To assess if the Rasch-scaled KIDSCREEN-52 generic health-related quality of life measure was valid in children with cerebral palsy (CP).

METHODS

The Rasch measurement properties and differential item functioning (DIF) of the KIDSCREEN-52 were examined in children with CP. Data were available from the KIDSCREEN project from 3219 children aged 8 to 12 years and 2126 parents in the general population; and from the SPARCLE project from 501 children aged 8 to 12 years with CP and 823 parents. Analysis used Zumbo's logistic regression DIF approach. Partial credit model analyses were conducted.

RESULTS

All items of the KIDSCREEN self-report version fitted the partial credit model (smallest P-value: 0.256). Only one item of the parent version did not fit the data well (smallest P-value 0.001). Statistically significant DIF was observed in some items, but was of substantial magnitude (ΔR2 = 0.046, 0.049) for only two items in two dimensions of the parent version. The practical impact of DIF was small. DIF-adjusted standardized mean differences between children with and without CP being 1.07 and 0.34 for the physical and school dimensions, respectively (unadjusted: 1.09 and 0.16).

CONCLUSIONS

The KIDSCREEN-52 functions in a similar way in children with CP and in the general population. Comparisons of quality of life between such children are therefore likely to be valid.

We have identified a cellular efflux pump, RhT, with the properties of an MDR transporter-a type of ATP-binding cassette transporter whose substrates include small hydrophobic molecules. RhT transports rhodamine 123 (Rh123) and is inhibited by low temperature, energy poisons, and several MDR transport inhibitors, such as verapamil. All vegetative cells have RhT activity, but during development prestalk cells lose RhT activity while prespore cells retain it. We also identified several RhT inhibitors. The most effective inhibitor is the stalk cell-inducing chlorinated alkyl phenone, DIF-1. The RhT inhibitors disrupted development, to varying degrees, and induced stalk cell formation in submerged culture. The inhibitors displayed the same rank order of pharmacological efficacy for stalk cell induction as they did for Rh123 transport inhibition. We also found that cerulenin, a specific inhibitor of DIF-1 biosynthesis (R. R. Kay, 1998, J. Biol. Chem. 273, 2669-2675), abolished the induction of stalk cells by each of the RhT inhibitors, and this effect could be reversed by DIF-1. Thus, DIF-1 synthesis appears to be required for the induction of stalk cells by the RhT inhibitors. Since DIF-1 is the most potent inhibitor of RhT activity, and thus a likely transport substrate itself, we propose that RhT inhibitors induce stalk cell differentiation by blocking DIF-1 export, causing DIF-1 to build up within cells. Our results provide evidence for a prespore-specific efflux pump that regulates cell fate determination, perhaps by regulating the cellular concentration of DIF-1.

We prospectively evaluated the efficacy of two commercial rapid methods for antigenic detection, a dot-blot enzyme immunoassay (EIA-DB) (Directigen FluA, Becton-Dickinson, USA) and a direct immunofluorescence assay (DIF) (Monofluokit Influenza A, Diagnostics Pasteur, France), compared with the shell-vial culture in the MDCK line, incubated 2 to 3 days and stained with the monoclonal antibody clone IA-52, the diagnosis of lower respiratory tract caused by Influenza A virus (IA). In the study period the presence of IA virus was detected in 59 of the 377 samples analyzed (15.7%). Only the SVC method detected all positive samples (100% sensitivity), being used as a reference method for comparison with the other techniques). The EIA-DB technique detected 50 cases (84.7%) and the DIF only 35 (59.3%). In nine (15.2%) cases the diagnosis was obtained only with the SVC method. The results of the comparison of the EIA-DB technique with SVC were: sensitivity 84.7%, specificity 100%, positive predictive value 100%, and negative predictive value 97.2%. The DIF technique gave values of 59.3%, 100%, 100%, and 92.9%, respectively. A statistically significant difference was observed between the sensitivity of the EIA-DB and the DIF method (p = 0.0001). In view of the results we recommended the use of the EIA-DB as a screening method when infection by the IA is suspected. But to obtain the maximum diagnostic yield all samples would be inoculated in a shell vial culture with the MDCK cell line.

BACKGROUND

Linear IgA bullous dermatosis (LABD) is an autoimmune subepidermal blistering disease that may be associated with drug exposure.

OBJECTIVE

Our purpose was to compare the clinical, pathologic, and immunofluorescence findings in drug-induced LABD with those in the idiopathic type.

METHODS

Six patients with an acute drug eruption were identified who had linear IgA deposition at the basement membrane zone (BMZ). Lesional tissue was examined by brightfield microscopy, and perilesional tissue was examined by direct immunofluorescence (DIF). The presence of circulating BMZ antibody was assayed by indirect immunofluorescence (IIF) on monkey esophagus (ME) and salt-split human skin (SS).

RESULTS

Histopathologic examination showed subepidermal bullae with varying numbers of inflammatory cells. DIF showed linear IgA at the BMZ; three of the patients also had weak deposition of C3 at the BMZ. Serum from five patients was studied by IIF. One patient had circulating IgA BMZ antibodies in a titer of 1:80 on ME, localized to the dermal side on SS. All patients were free of lesions within 5 weeks after discontinuation of the drug.

CONCLUSIONS

Drug-induced LABD is a self-limited eruption characterized by linear deposition of IgA without IgG at the BMZ. Most patients lack circulating antibodies. The distribution of lesions and the course of the disease differ from those of idiopathic LABD.

A novel differentiation-dependent cDNA (DIF-2) has been isolated from human mononuclear phagocytes by differential display. The full-length cDNA was cloned and sequenced. DIF-2 consists of 156 amino acids and has a predicted isoelectric point of 8.84. The mRNA is expressed in freshly isolated monocytes and is downregulated significantly when monocytes are subjected to differentiation. A similar differentiation-dependent downregulation is observed in normal hepatocytes compared to undifferentiated HepG2 cells. The mRNA expression in monocytes is sensitive to lipopolysaccharide and ceramide which both strongly increase DIF-2 transcription, while lysophosphatidylcholine results in a weaker upregulation of DIF-2 expression. A DIF-2 homologous gene has been previously isolated from mouse fibroblasts and was shown to be a serum growth factor-inducible immediate early gene. Our results indicate that DIF-2 represents a gene which is regulated in differentiation processes and strongly responsive to lipopolysaccharide, ceramide and lysophosphatidylcholine.

Dd-STATc becomes tyrosine phosphorylated, dimerises and accumulates in the nuclei of Dictyostelium cells exposed to DIF, the chlorinated hexaphenone that directs prestalk cell differentiation. By performing cytoplasmic photobleaching of living cells, we show that DIF inhibits the nuclear export of Dd-STATc. Within Dd-STATc there is a 50 amino acid region containing several consensus CRM1 (exportin 1)-dependent nuclear export signals (NESs). Deletion of this region causes Dd-STATc to accumulate in the nucleus constitutively and, when coupled to GFP, the same region directs nuclear export. We show that the N-terminal-proximal 46 amino acids are necessary for nuclear accumulation of Dd-STATc and sufficient to direct constitutive nuclear accumulation when fused to GFP. Combining the photobleaching and molecular analyses, we suggest that DIF-induced dimerisation of Dd-STATc functionally masks the NES-containing region and that this leads to nett nuclear accumulation, directed by the N-terminal-proximal import signals. These results show that the regulated nuclear accumulation of a STAT protein can be controlled at the level of nuclear export and they also provide a better understanding of the mechanism whereby DIF directs cell type divergence.