Stratified epithelium contains a mitotically active basal layer of cells that cease proliferating, then migrate outwards and undergo terminal differentiation. The control of this process, which is abnormal in cutaneous neoplasia and inf lammation, is not well understood. In normal epidermis, NF-B proteins were found to exist in the cytoplasm of basal cells and then to localize in the nuclei of suprabasal cells, suggesting a role for NF-B in the switch from proliferation to growth arrest and differentiation. Functional blockade of NF-B by expressing dominant-negative NF-B inhibitory proteins in transgenic murine and human epidermis produced hyperplastic epithelium in vivo. Consistent with this, application of a pharmacologic inhibitor of NF-B to intact skin induced epidermal hyperplasia. In contrast, overexpression of active p50 and p65 NF-B subunits in transgenic epithelium produced hypoplasia and growth inhibition. These data suggest that spatially restricted NF-B activation occurs in stratified epithelium and indicate that NF-B activation in this tissue, in contrast to its role in other settings, is important for cellular growth inhibition.

Tricho-rhino-phalangeal syndrome (TRPS) is characterized by craniofacial and skeletal abnormalities. Three subtypes have been described: TRPS I, caused by mutations in the TRPS1 gene on chromosome 8; TRPS II, a microdeletion syndrome affecting the TRPS1 and EXT1 genes; and TRPS III, a form with severe brachydactyly, due to short metacarpals, and severe short stature, but without exostoses. To investigate whether TRPS III is caused by TRPS1 mutations and to establish a genotype-phenotype correlation in TRPS, we performed extensive mutation analysis and evaluated the height and degree of brachydactyly in patients with TRPS I or TRPS III. We found 35 different mutations in 44 of 51 unrelated patients. The detection rate (86%) indicates that TRPS1 is the major locus for TRPS I and TRPS III. We did not find any mutation in the parents of sporadic patients or in apparently healthy relatives of familial patients, indicating complete penetrance of TRPS1 mutations. Evaluation of skeletal abnormalities of patients with TRPS1 mutations revealed a wide clinical spectrum. The phenotype was variable in unrelated, age- and sex-matched patients with identical mutations, as well as in families. Four of the five missense mutations alter the GATA DNA-binding zinc finger, and six of the seven unrelated patients with these mutations may be classified as having TRPS III. Our data indicate that TRPS III is at the severe end of the TRPS spectrum and that it is most often caused by a specific class of mutations in the TRPS1 gene.

IntroductionIn the present study, we analysed in detail nuclear autoantibodies and their associations in systemic sclerosis (SSc) patients included in the German Network for Systemic Scleroderma Registry.MethodsSera of 863 patients were analysed according to a standardised protocol including immunofluorescence, immunoprecipitation, line immunoassay and immunodiffusion.ResultsAntinuclear antibodies (ANA) were detected in 94.2% of patients. In 81.6%, at least one of the autoantibodies highly associated with SSc or with overlap syndromes with scleroderma features was detected, that is, anti-centromere (35.9%) or anti-topoisomerase I (30.1%), followed in markedly lower frequency by antibodies to PM-Scl (4.9%), U1-ribonucleoprotein (U1-RNP) (4.8%), RNA polymerases (RNAPs) (3.8%), fibrillarin (1.4%), Ku (1.2%), aminoacyl-transfer RNA synthetases (0.5%), To (0.2%) and U11-RNP (0.1%). We found that the simultaneous presence of SSc-associated autoantibodies was rare (1.6%). Furthermore, additional autoantibodies were detected in 55.4% of the patients with SSc, of which anti-Ro/anti-La, anti-mitochondrial and anti-p25/p23 antibodies were most frequent. The coexistence of SSc-associated and other autoantibodies was common (43% of patients). SSc-associated autoantibodies disclosed characteristic associations with clinical features of patients, some of which were previously not acknowledged.ConclusionsThis study shows that five autoantigens (that is, centromere, topoisomerase I, PM-Scl, U1-RNP and RNAP) detected more than 95% of the known SSc-associated antibody responses in ANA-positive SSc patients and characterise around 79% of all SSc patients in a central European cohort. These data confirm and extend previous data underlining the central role of the determination of ANAs in defining the diagnosis, subset allocation and prognosis of SSc patients.

Objective. Systemic sclerosis (SSc) is a rare, heterogeneous disease, which affects different organs and therefore requires interdisciplinary diagnostic and therapeutic management. To improve the detection and follow-up of patients presenting with different disease manifestations, an interdisciplinary registry was founded with contributions from different subspecialties involved in the care of patients with SSc.Methods. A questionnaire was developed to collect a core set of clinical data to determine the current disease status. Patients were grouped into five descriptive disease subsets, i.e. lcSSc, dcSSc, SSc sine scleroderma, overlap-syndrome and UCTD with scleroderma features.Results. Of the 1483 patients, 45.5% of patients had lcSSc and 32.7% dcSSc. Overlap syndrome was diagnosed in 10.9% of patients, while 8.8% had an undifferentiated form. SSc sine scleroderma was present in 1.5% of patients. Organ involvement was markedly different between subsets; pulmonary fibrosis for instance was significantly more frequent in dcSSc (56.1%) than in overlap syndrome (30.6%) or lcSSc (20.8%). Pulmonary hypertension was more common in dcSSc (18.5%) compared with lcSSc (14.9%), overlap syndrome (8.2%) and undifferentiated disease (4.1%). Musculoskeletal involvement was typical for overlap syndromes (67.6%). A family history of rheumatic disease was reported in 17.2% of patients and was associated with early disease onset (P < 0.005).Conclusion. In this nationwide register, a descriptive classification of patients with disease manifestations characteristic of SSc in five groups allows to include a broader spectrum of patients with features of SSc.

Summary Background  Intolerably high doses of systemic corticosteroids and additional immunosuppressants may be required to control disease activity in autoimmune bullous skin diseases. New therapeutic options are needed for such patients. Objectives  To determine the efficacy and adverse effects of adjuvant rituximab. Methods  Seven patients with refractory autoimmune blistering diseases (pemphigus vulgaris, PV, n = 4; bullous pemphigoid, BP, n = 2; mucous membrane pemphigoid, MMP, n = 1) were treated four times with rituximab at an individual dose of 375 mg m−2 at weekly intervals. Results  All lesions cleared in three patients (two PV, one BP), while they were reduced by more than 50% in three others (two PV, one BP). The concomitant immunosuppressive medication was reduced in five patients (four PV, one BP). The patient with MMP developed bilateral blindness while nasopharyngeal lesions resolved. Three patients (two BP, one PV) experienced severe adverse events including fatal pneumonia. Conclusions  Adjuvant B‐cell depletion by rituximab is effective in otherwise therapy‐resistant bullous autoimmune disorders but may be associated with substantial adverse effects including fatal outcomes.

Heat-shock protein (hsp) expression can be induced by high temperature, exposure to cytokines or oxygen radicals, ischemia, hemodynamic overload, or viral infections. To determine whether surface expression of hsp60 occurs in aortic endothelial cells stressed by high temperature or cytokines, cells from rat aortas were cultivated and stained with several types of monoclonal antibodies against hsp60. Other antibodies, eg, those against intercellular adhesion molecule-1 (ICAM-1), or immune response-associated antigens were also used as controls. Positive staining of endothelial cells on the surface and in the cytoplasm was observed after pretreatment of the cells with cytokine-containing medium, tumor necrosis factor-alpha (TNF-alpha), or interleukin-1 alpha and labeling with a specific monoclonal antibody against hsp60 (II-13). Fluorescence-activated cell sorter analyses showed that over 80% of living endothelial cells stressed by cytokine-containing medium, by TNF-alpha, or at 42 degrees C, but not by interleukin-1 alpha, were positively surface stained with this antibody. Increased intensity of immunostaining with antibodies to ICAM-1 and immune response-associated antigen was also seen on the cytokine-stressed endothelial cells. Furthermore, when TNF-alpha stimulated endothelial cells labeled with 51Cr were incubated with antibody II-13 in the presence of complement, significant lysis occurred. In summary, endothelial cells stressed by high temperature or certain cytokines, eg, TNF-alpha, express hsp60 in the cytoplasm and on their surfaces, and these cells were susceptible to complement-dependent lysis by hsp60-specific antibody. These observations may be significant for elucidating the mechanisms of the involvement of immune reactions to hsp65/60 in initiating atherosclerosis.

The evaluation of patients with contrast material-induced anaphylaxis (at least those with anaphylaxis > or = grade 2) should always include appropriate skin tests ensuring that patients with an IgE-mediated allergy are not missed. Moreover, allergologic testing may identify a contrast material of the group of nonionic monomers that will be tolerated in future radiologic interventions.

Figure 1Generation of epithelial cells with altered NF-κB function. Normal keratinocytes were transduced with retroviral expression vector for (a) lacZ normal control, (b) constitutively active p50, and (c) the trans-dominant IκBαM super-repressor, then subjected to immunofluorescence staining with antibody to p50 (bars = 5 µm). Note marked nuclear expression in p50-transduced cells and blockade of nuclear-localized p50 in IκBαM.NF-κB subunit transcription activation domains in that these changes are not observed with transcriptionally inactive mutants; the basis for this effect is unclear. This approach, however, has been shown previously to produce the predicted respective induction or blockade of NF-κB-driven gene expression in epithelial cells, both in the case of p50 and p65 alone as well as both subunits together (21) and provides a basis for determining the effects altering NF-κB function on apoptosis in this setting.Fas is expressed in epithelial cells and is implicated as a potentially important mediator of epithelial cell death in settings of inflammation (33-35) and ultraviolet injury (5,36). Cotransduction with vectors altering NF-κB function and a retroviral vector for human Fas/CD95 was used to study NF-κB effects on Fas-triggered apoptosis in epithelial cells. Fas activation in normal control and in IκBαM[+] cells leads to rapid cellular rounding, shrinkage, and detachment in the majority of cells; however, these changes are not seen in NF-κB subunit-expressing cells (Figure 2, a-c and f). These morphologic changes are consistent with apoptosis, a possibility supported by detection of DNA strand breakage using TUNEL assay and characteristic nuclear morphologic condensation and collapse in these cells (Figure 2, d and e).TNFα is another known trigger of apoptosis in many tissues that also impacts NF-κB function. TNFα activation of NF-κB through the TNF receptor TNFR1 opposes TNFα-induced apoptosis in a number of cell types in a process dependent on TRAF2 (8, 37). Whereas TNFα failed to alter normal keratinocytes or those expressing active NF-κB subunits, NF-κB blockade renders these cells very susceptible to TNFα-induced apoptosis (Figure 3, a-d), suggesting an analogous role for NF-κB in preventing apoptosis in epithelial cells.Blockade of NF-κB function leads to premature epidermal cell apoptosis in vivo. Normal stratified epithelium maintains a balance between cellular proliferation and a specialized form of programmed cell death confined to the outer differentiated cell layer at the stratum granulosum-stratum corneum interface (4). This terminal differentiation-associated cell death is not accompanied by classic cell morphologic features of apoptosis such as cell shrinkage and collapse, membrane blebbing, and nuclear condensation seen in other cell death settings in epithelium such as infection and ultraviolet injury (38). NF-κB translocates into nuclei of cells within outer layers of stratified epithelium (21), and this translocation is accompanied by NF-κB target gene activation (K. Hinata et...