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Biomedical subjects

U Neumann

Publications and source records attributed to U Neumann.

At least 109 records · Page 6Linked to original sources

Interleukin-4 promotes the expansion of skin-infiltrating lymphocytes from atopic dermatitis in vitro.

Functional studies of lymphocytes in atopic dermatitis (AD) have so far focused on peripheral blood mononuclear cells (PBMC), whereas cells at the involved site, the skin, have not been examined. Accordingly, we have developed methods to generate lymphocyte cultures from biopsies of inflammatory skin areas. Skin-infiltrating lymphocytes (SIL) were isolated from skin biopsies of 6 patients with severe AD and expanded in vitro in the presence of interleukin-2 (IL-2) without additional antigens. After 6-10 d in culture, outgrowth of mononuclear cells from biopsy tissue was observed in all cases. Phenotypic analysis of skin-derived cells revealed the predominance of CD4+ T-helper/inducer phenotype in SIL populations. Parallel cultures of SIL and PBMC showed an increase and expansion of CD8+ T cells in cultured PBMC, whereas the CD4+ phenotype was predominant in SIL cultures. As indicated by their expression of HLA-DR and CD25 antigens, most of the SIL were activated and the cells mainly expressed T-cell receptors (TCR) composed of alpha and beta chains. Different strategies for expansion of SIL in vitro were examined. High levels of IL-4 (1,000 U/ml) in combination with IL-2 (50 U/ml or 1,000 U/ml) preferentially promoted growth of SIL derived from AD and were much more effective than IL-2 alone. No cells expanded in cultures with IL-4 alone. SIL grown with high concentrations of IL-4 contained a significant proportion of double-positive CD4+8+ cells. No other marked differences were observed in the distribution of T cell subsets in cultures propagated under different conditions for 21 d. Our results demonstrate the feasibility of growing infiltrating T lymphocytes from inflammatory skin of AD patients. The use of high concentrations of IL-2 in combination with high levels of IL-4 allows a large expansion of these cells and thus represents a useful strategy to expand cells for further functional and molecular biologic studies.

Adult↗

Functional characterization of skin-infiltrating lymphocytes in atopic dermatitis.

Skin-infiltrating lymphocytes (SIL) were isolated from skin biopsies of patients with hyperimmunoglobulin E (IgE) atopic dermatitis (AD) and expanded in vitro in the presence of IL-2 in combination with IL-4. Phenotypic analysis of skin-derived cells revealed the predominance of CD4+ T helper/inducer phenotype in SIL populations. In 3H-thymidine incorporation assays, SIL showed proliferation in response to IL-2, IL-3, IL-4, ionomycin (Io) + 12-o-tetradecanoyl-phorbol-13-acetate (TPA) and OKT3 + TPA. OKT4 with and without TPA did not induce proliferation. Tumour necrosis factor alpha (TNF-alpha) did not block proliferative responses of SIL to IL-2 and IL-4. Cultured SIL showed no cytotoxic activity against K562 and Jurkat target cells. Expanded skin-derived T cells were tested for their capacity to secrete several cytokines in vitro. SIL secreted significant amounts of IL-4, GM-CSF and TNF-alpha upon stimulation with mitogens but failed to secrete IFN-gamma. Io in combination with phorbol-ester induced the secretion of larger amounts of IL-4, GM-CSF, TNF-alpha and low amounts of IFN-gamma. The data indicate that SIL derived from AD lesions were defective in their capacity to secrete IFN-gamma but were enriched in T cells capable of producing IL-4 upon stimulation. The results support the possibility of a predominant 'TH2-like' cell-mediated immune response in lesional skin of AD patients.

Antibodies, Monoclonal↗

Reaction of dipeptidylpeptidase IV with substrate-analogous azapeptides.

The reaction of dipeptidyl peptidase IV (EC 3.4.14.5.) with azapeptide substrates containing azaalanine or azaproline in the P1-position was investigated. Accumulation of a fairly stable acyl-enzyme could be shown for ester substrates. Ala-AzaPro-pNA is a very poor substrate of DP IV and does not accumulate an acyl-enzyme. DP IV does not react with active-site titrants for trypsin-like serine proteases.

Amino Acid Sequence↗

Inhibition of chymotrypsin and pancreatic elastase by 4H-3,1-benzoxazin-4-ones.

Eight 3,1-Benzoxazin-4-ones have been used to inactivate chymotrypsin and pancreatic elastase. Whereas 6,7-dimethoxy substitution only slightly decreased the acylation rate constant, the deacylation reaction was nearly unaffected. Bulky alkoxy groups in position 2 of the heterocyclic moiety were shown to increase enormously the acylation rate of chymotrypsin, but not that of elastase.

Acylation↗

["Ledderhose" disease. Plantar fibromatosis--clinical aspects].

The Ledderhose's diseases is a relatively rare and not well known clinical picture. Even there are tight pathomorphological and clinical relations to the Dupuytren's contracture, the genesis is also here quite unknown. Because of inefficiency of conventional therapy the surgical treatment is the only alternative. On the sample of 12 operations in 7 patients from 1979 to 1989 surgical procedure and long-term results are discussed.

Adult↗

[The occurrence of Campylobacter spp. and Salmonella spp. in gulls in northern Germany].

Out of a total of 207 gulls--20 Black headed gulls, 185 Herring gulls and 2 Common gulls--128 (62%) and 23 (11%) birds, respectively, were infected with Campylobacter spp. (C.) and Salmonella spp. C. jejuni was predominant in gulls less than two years old (89%) and C. coli in older birds (75%). Furthermore, the infection rate with Campylobacters was depending on the habitats of the birds. The rate for C. jejuni and C. coli, respectively, was in gulls from regional garbage dumps 78% and 4%, from the coast 58% and 21%, and from islands 47% and 47% of the isolations in the corresponding area. Salmonellae were mainly isolated in the period from September to February from gulls less than one year old and from birds from the coast.

Animals↗

Effect of antipyrine and phenobarbital on renal gamma-glutamyltransferase excretion in human urine.

Serum gamma-glutamyltransferase is used as a marker of hepatic enzyme induction. The kidney contains high activities of gamma-glutamyltransferase in the brush border membrane of the proximal tubule, from which it is released into urine. This study investigated the effect of phenobarbital and antipyrine, two inducers of hepatic monoxygenases and gamma-glutamyltransferase, on the urinary excretion of renal gamma-glutamyltransferase. Three groups (n = 6) of healthy male volunteers received 100 mg phenobarbital for 7 and 14 days and 1200 mg antipyrine for 7 days, respectively. Antipyrine and phenobarbital increased antipyrine elimination, serum gamma-glutamyltransferase, and the urinary excretion of renal gamma-glutamyltransferase, whereas urinary beta-N-acetylglucosaminidase, beta-glucuronidase, and total protein and glucose excretion were unchanged. No correlation was found between serum and urinary gamma-glutamyltransferase or both enzymes and antipyrine elimination. Increases in antipyrine elimination were positively correlated to increases in serum, but not urinary gamma-glutamyltransferase. The findings suggest that antipyrine and phenobarbital increase urinary gamma-glutamyltransferase excretion. However, the increase in urinary gamma-glutamyltransferase does not reflect the magnitude of hepatic enzyme induction.

Adolescent↗

Modified photometric method for the determination of phospholipase A activities.

A modified photometric method for the determination of phospholipase A activities which is based on a previously published reaction principle is described. The modified assay uses a lyophilized substrate emulsion rather than a freshly prepared phospholipid emulsion and a new chromogen (tribromohydroxybenzoic acid and 4-amino-antipyrine) with a high molar absorption coefficient (1,930 m2/mol at 546 nm, 2,900 m2/mol at 512 nm wavelength). The new test is more practicable with respect to pipetting volumes and incubation times. Preliminary results of a method evaluation indicate that the modified assay fulfills the usual criteria for clinical chemical enzyme measurements.

Humans↗

Reactions between dipeptidyl peptidase IV and diacyl hydroxylamines: mechanistic investigations.

Kinetics of inactivation of dipeptidyl peptidase IV (DP IV, EC 3.4.14.5) by N-peptidyl-O-(4-nitrobenzoyl) hydroxylamines and their enzyme-catalyzed hydrolysis were followed using independent monitoring methods, all giving similar efficiency ratios of Kcat/Kinact. Different temperature dependences of the DP IV-inactivation and enzyme-catalyzed hydrolysis provide evidence of independent rate determining steps for both reactions. Activation parameters of inactivation are similar to those of spontaneous decomposition of the compounds, suggesting a mechanistic relationship. Investigation of DP IV-inactivation, DP IV-catalyzed hydrolysis of N-Ala-Pro-O-Bz(4-NO2) and the decomposition of the suicide substrate in H2O and D2O gave solvent isotope effects of 4.65, 2.54 and 1.02, respectively. A proton inventory of the inactivation reaction indicates involvement of more than one proton in the formation or breakdown of its transition state. The linear proton inventory found for the hydrolytic reaction is consistent with one proton transition in the rate determining step and resembles the rate limiting deacylation of Ala-Pro-DP IV. The hypothetical reaction model now locates splitting in both reactions prior to formation of a covalent intermediate during the catalytic cycle.

Dipeptidyl Peptidase 4↗

[The occurrence of paramyxoviruses in game birds: epizootiological aspects, characteristics in vivo and in vitro. A review of the literature].

The recent review is based on recent literature on the epizootiology and on properties of avian paramyxoviruses in vivo and in vitro. Paramyxoviruses are classified according to their pathogenicity (especially serotype 1), in addition to serotyping by means of hemagglutination-inhibition tests and by monoclonal antibody binding assays. Also, neuraminidase-inhibition tests, double-immunodiffusion tests can be performed. Growth properties of the different paramyxovirus-serotypes are accessory techniques to classify paramyxoviruses parallel to the determination of the pathogenicity. Structural polypeptides can be demonstrated in polyacrylamide-electrophoresis, thus leading to a distinction of the different serotypes.

Animals↗

[Comparative studies of paramyxovirus isolates from wild birds].

As a result of a surveillance programme in North-Germany, paramyxovirus-isolates of serogroup 1 with different pathogenicity were isolated from different species of feral birds (Black-headed gull, mallard, tawny owl, tree sparrow, mute swan). In chick embryo fibroblast monolayers these isolates produced plaques exhibiting different plaque morphology. After cloning the isolates, further physicochemical, serological and pathogenicity testings were performed. On the basis of mean death time and intracerebral pathogenicity index, two of the isolates were found to be avirulent (lentogenic), whereas the others proved to be virulent (velogenic). SDS-Polyacrylamide-gel-electrophoresis showed polypeptide patterns similar to that of the PMV-1 reference strains. Summarizing all relevant findings of these investigations under consideration of the suggestions for the virological description of avian PMV (Alexander, 1987; Alexander et al., 1987; Who-Expert-Committee, 1980), including the recently established serotyping by means of monoclonal antibodies, the following designations of the PMV isolates tested herein are proposed: 1. PMV-1/black-headed gull/Germany/SSP-233/83, lentogenic NDV, monoclonal antibody group G 2. PMV-1/mallard/Germany/SSP-284/83, lentogenic NDV, monoclonal antibody group H 3. PMV-1/tawny owl/Germany/SSP-262/84, velogenic NDV, monoclonal antibody group B 4. PMV-1/tree sparrow nestling/Germany/SSP-352/84, velogenic NDV, monoclonal antibody group B 5. PMV-1/mute swan/Germany/SSP-204/85, velogenic NDV, monoclonal antibody group B.

Animals↗

The rate of Wallerian degeneration in the absence of immunoglobulins. A study in chick and mouse peripheral nerve.

Sciatic nerves undergoing Wallerian degeneration are subject to massive invasion by monocytes bearing Fc receptors. The present experiments were done to explore the potential role of the Fc receptors. The effect of suppressed immunoglobulin levels on the rate of Wallerian degeneration was studied in the n. medioulnaris of the chick and in murine sciatic nerve. In the avian model, total immunoglobulin production was eliminated by bursectomy. The mice were injected after birth with antibodies against IgM, causing a selective reduction of immunoglobulin M levels. Myelin degeneration was measured in both groups by computer-assisted line sampling and corrected for intrafascicular edema. In both models, there were only minor differences with the controls. Immunoglobulins, accordingly, do not seem to play a significant role in the efficiency of myelin phagocytosis by monocytes during Wallerian degeneration. The study also includes data on the degree and the distribution of edema in nerves undergoing Wallerian degeneration.

Animals↗

Enzymic properties of intestinal aminopeptidase P: a new continuous assay.

A continuous photometric assay of aminopeptidase P activity was developed which is based on a coupled enzymic assay with the substrate Gly-Pro-Pro-pNA and DPP IV as auxiliary enzyme. This assay was used to evaluate the kinetic parameters and inhibitory profile of intestinal brush border aminopeptidase P.

Amino Acid Sequence↗

P300 and anticipated task difficulty.

Recently, it has been shown that P300 measures depend on stimulus evaluation processes. This paper explores the effect of messages informing about the degree of difficulty of subsequent tasks on the P300 component. Ten healthy volunteers were asked to solve mental arithmetic tasks of graduated difficulty. The degree of difficulty was announced by messages which preceded the tasks and which were used as P300 eliciting stimuli. Measurements of the amplitude and latency of P300 components from 3 brain regions (Fz, Cz, Pz) were collected selectively for each category of task difficulty announced. Subjective ratings of difficulty and the frequency of errors confirmed the monotonic increase of task difficulty from categories I to VI. The mean P300 amplitude in categories I-VI showed a U-shaped trend, i.e. the highest amplitude occurred in the extreme categories 'extremely easy' and 'very difficult'. No significant effect of stimulus categories on P300 peak latencies was found. It is concluded that the P300 amplitude reflects an evaluation in the sense of distance judgements on an internal scale which in this study concerned the task difficulty.

Adult↗