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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    British journal of dermatology 145 (2001), S. 0 
    ISSN: 1365-2133
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background The frequency and amount of tissue eosinophilia in spontaneous lesions of acute and chronic atopic dermatitis (AD) are still a matter of controversy, and little is known about the distribution of eosinophilia in skin. Objectives To give a quantitative description of tissue eosinophilia in spontaneous lesions of acute and chronic AD based on morphometric data. Methods Thirty-one lesional skin biopsies of AD were evaluated using our recently described method for the quantitative assessment of eosinophilic granule protein (EGP) deposition by image analysis of immunostaining using the antibodies EG1, EG2, MBP, EPO and neutrophil elastase (NE). The frequency, amount and distribution of protein deposition including extracellular EGP deposition as an indicator of complete activation and degranulation of eosinophils were determined. Eosinophil count was performed in addition. Histopathological parameters of acute dermatitis (spongiosis) and chronic dermatitis (epidermal hyperplasia) were scored to look for a correlation with tissue eosinophilia. Results Tissue eosinophilia was found in nearly all biopsies (30 of 31). The most protein was detected by EG2, followed by EG1, MBP and EPO, with very small amounts of NE. A superficial tissue distribution of eosinophilia was found, with 〈 10% of total EGP deposition below a depth of 1·39 mm from the epidermis. Eosinophils were involved in acute, spongiotic dermatitis, but more tissue eosinophilia including EGP deposition was detected in lesions with pronounced epidermal hyperplasia than in biopsies without. Conclusions These data provide further evidence for the involvement of activated eosinophils in acute and chronic AD by a new quantitative in situ approach. Pronounced tissue eosinophilia, especially EGP deposition as the result of complete activation of eosinophils, is found in chronic AD and may be involved in the development or maintenance of chronicity.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    British journal of dermatology 142 (2000), S. 0 
    ISSN: 1365-2133
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The histopathological differentiation between well-differentiated carcinomas and atypical adenomas of sweat gland origin may be difficult, even if immunohistochemical methods are used. Therefore, additional techniques may be helpful. We previously demonstrated that DNA image cytometry (ICM-DNA) can be useful in distinguishing between malignant and benign clear cell hidradenoma. In the present study, a larger series of sweat gland tumours, with a clear-cut diagnosis as malignant or benign on histopathological criteria, was examined by ICM-DNA. Enzymatic cell separation specimens were prepared from paraffin-embedded tissues of 18 sweat gland carcinomas (14 porocarcinomas, one classic eccrine adenocarcinoma, two microcystic adnexal carcinomas and one mostly ductal apocrine carcinoma) and 47 benign sweat gland tumours (three syringocystadenomas, five spiradenomas, 14 cylindromas, three syringomas, seven nodular hidradenomas, 10 cutaneous mixed tumours, four poromas and one apocrine hidrocystoma). Specimens were examined by ICM-DNA according to the current recommendations of the European Society for Analytical Cellular Pathology with the AutoCyte QUIC-DNA workstation using mesenchymal cells as an internal reference. DNA aneuploidy was detected by the stemline interpretation according to Böcking and/or at least three 5[c]-exceeding events. DNA aneuploidy was detected in 16 of 18 (89%) of the sweat gland carcinomas, but in none of the 47 adenomas. These results suggest that the detection of DNA aneuploidy in sweat gland tumours using ICM-DNA is a clear and specific indicator of prospective malignancy.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    British journal of dermatology 142 (2000), S. 0 
    ISSN: 1365-2133
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: In patients with atopic dermatitis two different types of blood eosinophils with distinct density can be isolated. The normodense cells represent the huge majority in count, whereas the hypodense eosinophils are characterized by higher effector activity. To understand the altered functional responsiveness of these two cell subtypes, the expression of C5a receptors as well as C5a-induced signal pathways and the production of reactive oxygen metabolites have been analyzed. Chemiluminescence measurements revealed significant higher production of reactive oxygen metabolites in hypodense eosinophils in comparison to normodense cells. However, no difference in the expression level of C5a receptors as well as in the C5a-induced Ca2+-transients between normodense and hypodense eosinophils were found. In contrast, hypodense eosinophils showed a significantly higher actin polymerization response and phosphatidylinositol 4,5 bisphosphate 3-kinase activation after stimulation with C5a than normodense eosinophils. Therefore, normodense and hypodense eosinophils from the blood of patients with atopic dermatitis are characterized by differential amplification of C5a-receptor signal pathways, which might explain the differences in their proinflammatory activity.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Pediatric allergy and immunology 4 (1993), S. 0 
    ISSN: 1399-3038
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: In order to study cell activation in peripheral blood on bronchial allergen provocation up to 24 h, we investigated 32 asthmatic children, sensitive to house-dust mites. Six healthy young adult volunteers served as controls. Lymphocyte subsets (CD3, CD19, CD4, CDS) and activation markers (CD25-T, HLADR-T, CD23) in peripheral blood as well as soluble IL2-R and soluble ICAM-1 in scrum were evaluated. In terms of clinical reaction, 23 children exhibited a DAR, 6 an EAR, 6 a LAR and 3 children did not show a bronchoconstrictor response to allergen challenge with house-dust mite extract (NAR). In comparison to controls, asthmatic children showed a significantly higher expression of CD23 on B-lymphocytcs (p 〈 0.05). Other subsets were in the same range in both groups. After provocation there was a significant increase of CD4/CD8-ratio only in asthmatic children. Serum levels of sIL2-R were significantly higher in asthmatic children compared to controls at baseline as well as at 12 and 24 h after provocation, without variation during observation period, No differences were noted for SICAM-1. Our results confirm the hypothesis that lymphocytes, as important cells in regulation of allergic immune response, are recruited into peripheral blood under allergen challenge conditions in sensitized asthmatic children.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Clinical & experimental allergy 34 (2004), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Different chemokine receptors have been suggested to play a pivotal role in allergic diseases and therefore to be relevant for the activation of effector cells and propagation of the inflammatory response. The CXC chemokine receptor CXCR4 has recently been found on the surface of eosinophils implicating a role in allergic diseases.Objective The aim of this study was to investigate the functional expression of CXCR4 on senescent eosinophils. Moreover, we questioned whether the cytokine profile – T-helper (Th)1 and Th2 cytokines – affect the activation of eosinophils via the CXCR4 that could be important for the different phases of the allergic reaction.Methods CXCR4 expression on human eosinophils was analysed by flow cytometry and RT-PCR. Functional analyses of intracellular calcium fluxes, actin polymerization, release of reactive oxygen species and, chemotaxis were carried out using spectrofluorometry, flow cytometry, chemiluminescence and modified Boyden chamber technique.Results Whole blood and freshly isolated eosinophils weakly express CXCR4 surface protein. Incubation in culture medium without addition of cytokines for 24 h always lead to strong CXCR4 surface expression that paralleled with stromal-derived factor-1α (CXCL12)-induced eosinophil activation. Stimulation of eosinophils with CXCL12 leads to an internalization of CXCR4, which could be prevented by phenylarsine oxide. Co-incubation of eosinophils with Th2 cytokines such as IL-3, IL-4, IL-5, IL-13, and granulocyte macrophage-colony stimulating factor prevented the expression of CXCR4 and affected eosinophil activation after stimulation with the CXCR4 ligand CXCL12. From these cytokines, IL-3 was the only cytokine completely inhibited intracellular calcium fluxes and chemotaxis of eosinophils in response to CXCL12.Conclusion Senescent eosinophils express functional CXCR4 receptors, which are prevented by Th2 cytokines that are found in the early phase of allergic reaction. Therefore, CXCR4 activation of eosinophils seems to be important in the chronic phase of allergic reaction, which is dominated by a Th1 cytokine profile.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Staphylococcus aureus is a well known trigger factor of atopic dermatitis (AD). Besides the superantigens, further exotoxins are produced by S. aureus and may have an influence on the eczema.Objective To explore the impact of staphylococcal α-toxin on human T cells, as those represent the majority of skin infiltrating cells in AD.Methods Adult patients with AD were screened for cutaneous colonization with α-toxin producing S. aureus. As α-toxin may induce necrosis, CD4+ T cells were incubated with sublytic α-toxin concentrations. Proliferation and up-regulation of IFN-γ on the mRNA and the protein level were assessed. The induction of t-bet translocation in CD4+ T cells was detected with the Electrophoretic Mobility Shift Assay.Results Thirty-four percent of the patients were colonized with α-toxin producing S. aureus and α-toxin was detected in lesional skin of these patients by immunohistochemistry. Sublytic α-toxin concentrations induced a marked proliferation of isolated CD4+ T cells. Microarray analysis indicated that α-toxin induced particularly high amounts of IFN-γ transcripts. Up-regulation of IFN-γ was confirmed both on the mRNA and the protein level. Stimulation of CD4+ T cells with α-toxin resulted in DNA binding of t-bet, known as a key transcription factor involved into primary T helper type 1 (Th1) commitment.Conclusion α-toxin is produced by S. aureus isolated from patients with AD. We show here for the first time that sublytic α-toxin concentrations activate T cells in the absence of antigen-presenting cells. Our results indicate that α-toxin is relevant for the induction of a Th1 like cytokine response. In AD, this facilitates the development of Th1 cell dominated chronic eczema.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Clinical & experimental allergy 34 (2004), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Food allergy is a common problem in patients with atopic dermatitis (AD), particularly in children. While immediate reactions to food are well characterized, the importance of food as a provocation factor for late eczematous reactions has been a subject of debate for several decades.Objective To investigate the importance of food for the induction of late eczematous reactions in children with AD and to correlate the clinical outcome to the results of specific IgE determinations and atopy patch tests (APTs).Methods One hundred and six double-blind placebo-controlled food challenges (DBPCFCs) to cow's milk, hen's egg, wheat and soy in 64 children with AD (median age 2 years) were analysed retrospectively. Total and food-specific IgE were determined by CAP RAST FEIA and APTs with native foodstuff were performed. The diagnostic values of specific IgE and APT results were calculated.Results Forty-nine (46%) of the challenges were related to a clinical reaction. An exacerbation of AD (late eczematous reaction) commonly occurred 24 h after the ingestion of food. Isolated late eczematous reactions were seen in 12% of all positive challenges. Forty-five percent of the positive challenges were associated with late eczematous responses, which followed immediate-type reactions. The sensitivity of food-specific IgE and the APT was 76% and 70%, respectively. Specific IgE and APT were often false positive, which resulted in low positive predictive values (64% and 45%, respectively).Conclusions Late eczematous reactions may often be observed upon food challenge in children with AD. Due to the poor reliability of food-specific IgE and APT results DBPCFCs have still to be regarded as the gold standard for the appropriate diagnosis of food responsive eczema in children with AD.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 26 (1996), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Bovine casein leads to an expansion of lymphocytes expressing the cutaneous lymphocyte antigen and to specific lymphocyte proliferation in a subgroup of patients with milk-responsive atopic dermatitis (AD). The casein fraction is composed of different proteins with defined and completely different sequences.Objective To define the stimulatory capacity of the major casein protein (α and k) in lymphocyte proliferation assays with cells from milk-allergic and non-allergic individuals.Methods Proliferative responses of peripheral blood mononuclear cells to lipopolysaccharide-depleted casein subfractions were measured by thymidine incorporation. Lymphocytes from milk-responsive patients with AD were compared with cells from non-responsive patients with AD and to non-atopic individuals, Atopic individuals with immediate symptoms following consumption of cow's milk were included as positive controls. Casein-specific T-cell clones (TCC) from four patients with milk-responsive AD were restimulated with unfractioned casein and K-casein.Results Higher proliferative responses to unfractionated casein and α-,β and casein were observed in milk-responsive patients compared with non-responders. Unfractionated casein and K-casein discriminated best between the milk-responsive patients with AD and non-responders. Twenty-five of 31 TCC from patients with milkresponsive AD reacted to the mixed casein preparation and K-casein.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 25 (1995), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Type of Medium: Electronic Resource
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