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  • 1
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The N-terminal fragment, comprising residues −5 to 55 of herpes simplex virus type 1 glycoprotein D was expressed as a β-galactosidase fusion protein inEscherichia coli. This gD-fusion protein reacts with monoclonal antibody LP14 directed against glycoprotein D of HSV. Antisera obtained after immunization of rabbits with purified gD-fusion protein react with HSV-1 gD in a Western blot and with N-terminal synthetic peptides of gD. In addition, these antisera are able to neutralize viral infectivity in vitro.
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  • 2
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary To locate T cell determinants of glycoprotein D (gD) of herpes simplex virus type 1 (HSV-1), proliferation assays of lymphocytes obtained from 10 healthy HSV-seropositive individuals were performed using 34 overlapping gD peptides as antigens. Despite large differences between individual responses to the peptides both in number of stimulating peptides and gD regions, three regions (1–54, 110–214, and 290–314) induced a response in 50% or more of the HSV-seropositives. T cells were less frequently stimulated by peptides of region 210–294. No correlation was found between serological data and proliferative responses to the peptides. The diversity in T cell response to the peptides suggests a lack of immunodominance, implying that a single peptide/region of gD, or a combination of peptides, will not be sufficient to serve as a basis for a future HSV-1 vaccine.
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  • 3
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Several analogues of the amino acid sequence of peptide 9–21 of glycoprotein D of herpes simplex virus type 1 (HSV-1) were synthesized and investigated for reactivity with different group VII monoclonal antibodies, Mabs LP14, ID3, 170, HD4, A16, EII-24 and EV-10, in a competition enzyme-linked immunosorbent assay (ELISA). Replacement of Arg at position 16 by His resulted in a loss of binding with the group VII Mabs. Substitution of Pro at residue 14 by Leu gave a reduced binding for a number of Mabs and loss of binding for Mab A16. Substitution of Lys at position 10 by Glu gave reduced binding for three out of the seven Mabs. In addition substitutions of Met at position 11 by norleucine and oxidized Met were studied. The boundaries of the epitope cluster were mapped by studying synthetic variants of peptide 9–21 which were shorter either at the C-terminus or at the N-terminus, or both. Peptide 10–18 and peptide 9–17 were the shortest peptides, which were still reactive with the group VII Mabs. Mab HD4 requires the N-terminus of peptide 9–21 for effective binding, while for binding of other Mabs contribution of the residues in the C-terminal part of this peptide is more important.
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  • 4
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Thirty eight human sera, seropositive for herpes simplex virus (HSV) and 56 human sera, seronegative for HSV by immunofluorescence and by ELISA, were investigated for reactivity with a series of overlapping synthetic peptides of HSV type 1 glycoprotein D (gD-1). Thirty four out of the 38 human sera positive for HSV reacted with peptides located between residues 300 and 369; the HSV-negative sera reacted with six of the gD-1 peptides, but with none of the peptides within residues 300 to 369.
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  • 5
    ISSN: 1573-739X
    Keywords: Ciprofloxacin ; Colonization resistance ; Faecal flora ; Norfloxacin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Oral and faecal flora were monitored in ten healthy volunteers who received three different oral dosages of either norfloxacin or ciprofloxacin during five consecutive days. Oral flora was not influenced by either quinolone. At the end of the five day-treatment period aerobic Gram-negative bacilli could no longer be cultured from the faecal samples of any of the volunteers. Enterococci were suppressed in some volunteers or remained present in concentrations as high as before treatment. All ciprofloxacin-treated volunteers became colonized with yeasts in their faecal samples, even during treatment with the lowest dose. As measured by total microscopic bacterial clump counts and the appearance of ß-aspartylglycine in faecal samples neither of the two quinolones appeared to decrease the colonization resistance of the digestive tract.
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  • 6
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background It has been suggested that intestinal microbiota of allergic and non-allergic children differs in composition, and that microbiota–immune system interactions may predispose children to develop sensitization. Previous studies have examined fecal microbiota of allergic children with atopic dermatitis, but little is known about that of atopic wheezy children.Objective To investigate the composition of the fecal microbiota of young sensitized wheezy and non-sensitized non-wheezy children, using molecular methods.Methods Within the context of a prospective birth cohort, we carried out a nested case–control study of sensitized wheezy children (cases) and non-sensitized non-wheezy controls. Cases and controls were matched for age, sex, parental atopy, allergen exposure, and pet ownership. We evaluated the composition of fecal microbiota by nucleic acid-based methods (PCR combined with denaturing gradient gel electrophoresis and quantification of bifidobacteria by fluorescent in situ hybridization).Results Thirty-three case–control pairs (mean age 4.4 years) provided stool samples. Comparison of total bacterial community profiles showed that each child had a unique fecal microbiota (mean Dice's similarity coefficient 22%, range 3.3–60.8%). There was no difference between the groups in prevalence of Lactic Acid bacteria (12/33 vs. 11/33, P=0.8) or bifidobacteria (30/33 vs. 31/33, P=1.00, cases vs. controls). The bifidobacterial species detected were similar in both groups. The percentage of bifidobacteria in total fecal microflora was no different between cases (median 1.7%, range 0–20.8%) and controls (1.9%, 0–18.2%, P=0.7). However, cases with eczema had significantly fewer bifidobacteria (median 1.6%, range 0–4.8%) than their controls (4.0%, 1.9–18.2%, P=0.05).Conclusion We found no differences in fecal microbiota composition between sensitized wheezy and non-sensitized, non-wheezy children aged 3–5 years using nucleic acid-based methods. Differences appear to be isolated to those allergic children with eczema.
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  • 7
    ISSN: 1573-4927
    Keywords: ribonuclease ; amino acid sequence ; polymorphism ; Camelidae ; pancreas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Pancreatic ribonucleases from several species (whitetail deer, roe deer, guinea pig, and arabian camel) exhibit more than one amino acid at particular positions in their amino acid sequences. Since these enzymes were isolated from pooled pancreas, the origin of this heterogeneity is not clear. The pancreatic ribonucleases from 11 individual arabian camels (Camelus dromedarius) have been investigated with respect to the lysine-glutamine heterogeneity at position 103 (Welling et al., 1975). Six ribonucleases showed only one basic band and five showed two bands after polyacrylamide gel electrophoresis, suggesting a gene frequency of about 0.75 for the Lys gene and about 0.25 for the Gln gene. The amino acid sequence of bactrian camel (Camelus bactrianus) ribonuclease isolated from individual pancreatic tissue was determined and compared with that of arabian camel ribonuclease. The only difference was observed at position 103. In the ribonucleases from two unrelated bactrian camels, only glutamine was observed at that position.
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  • 8
    ISSN: 1052-9306
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A peptide comprising 37 amino acids of the antigen binding site of a monoclonal antibody directed against glycoprotein D of herpes simplex virus was synthesized. The synthetic peptide and the impurities formed in the synthesis were characterized by capillary electrophoresis/ionspray mass spectrometry and by 252Cf plasma desorption-time of flight mass spectrometry. The measured average molecular mass of the synthetic peptide was 4627.16 Da, which was only 0.08 Da higher than the calculated value (4627.08 Da). The plasma desorption mass spectrum of the synthetic peptide showed a protonated molecule at m/z 4624.1, which was 4 Da lower than the calculated one (4628.09 Da). The amino acid sequence of the peptide was confirmed in part by electrospray (ionspray) mass spectrometry using a high nozzle skimmer voltage difference. Five impurities were separated and identified by capillary electrophoresis/mass spectrometry and two of them also appeared in the plasma desorption mass spectrum.
    Additional Material: 6 Ill.
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  • 9
    ISSN: 1439-0973
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Bei 25 gesunden Probanden wurde eine Untersuchung zur Inaktivierung von Aztreonam durch die Stuhlflora durchgeführt. Zehn Probanden erhielten Aztreonam oral verabreicht, 15 Probanden erhielten kein Antibiotikum. Mittels HPLC wurden simultan Bestimmungen des restlichen Aztreonam und seiner inaktivierten offenen Ringform vorgenommen. In der Stuhlflora von 20 Probanden war unabhängig von der Aztreonam-Gabe β-Laktamase-Aktivität nachzuweisen. Die Inaktivierung von Aztreonam durch den Überstand von Stuhlsuspensionen ließ sich durch Clavulansäure hemmen.
    Notes: Summary The enzymatic inactivation of aztreonam by the faecal flora was investigated in 25 healthy human volunteers. Ten volunteers received aztreonam orally and 15 volunteers did not receive antibiotic treatment. Residual aztreonam and its inactivated open ring form were simultaneously detected by an HPLC-assay. Independent of aztreonam treatment, β-lactamase activity was detected in the faecal flora of 20 volunteers. The inactivation of aztreonam by the faecal supernatants was inhibited by clavulanic acid.
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  • 10
    ISSN: 1439-0973
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Die normale Flora des Intestinaltraktes besteht vorweigend aus anaeroben Bakterien und schützt den Wirt gegen eine Kolonisation durch pathogene Mikroorganismen. Eine antimikrobielle Therapie mit Ceftriaxon kann die Mikroflora des Dickdarms beeinträchtigen und damit auch deren protektiven Effekt. Zehn gesunde Probanden erhielten fünf Tage lang 1 g Ceftriaxon intramuskulär appliziert. Dies führte zu einer signifikanten Abnahme der mittleren Koloniebildnerzahlen von 10,67 (SEM ± 0,11) vor Applikation auf 9,02 (± 0,45) nach drei und auf 8,97 (± 0,46) nach fünf Tagen (p〈0,05). Nach zehn und 15 bis 19 Tagen im Anschluß an die Antibiotikagabe kehrten die Anaerobier-Koloniebildnerzahlen auf 10,17 (± 0,16) bzw. 10,44 (± 0,18) zurück. Bakterienenzyme können als Indikator für die protektive Mikroflora dienen. In Überständen von Stuhlproben der Probanden wurden β-Aspartylpeptidase und Desoxycholat-Hydrolase bestimmt und mit den Anaerobier-Kulturen verglichen. Zwischen den Aktivitäten beider Enzyme und der am Tag 3 gemessenen Anaerobier-Gesamtzahl fand sich eine signifikante Korrelation. Am Tag 5 und Tag 8 zeigte nur die β-Aspartylpeptidase eine signifikante Korrelation mit den Gesamt-Kolonie-bildnerzahlen der Anaerobier sowie mit den Zahlen vonBacteroides spp. oder Bifidobakterien. An den Tagen 15 bis 19 (zehn bis 14 Tage nach Antibiotikagabe) bestand nur zwischen der Zahl vonBacteroides spp. und β-Aspartylpeptidase eine signifikante Korrelation. Nach Behandlung mit Ceftriaxon lassen sich folglich Veränderungen der bakteriellen Flora kurzfristig durch Bestimmung der β-Aspartylpeptidase erfassen, weniger gut aber die Erholung der Darmflora.
    Notes: Summary The normal flora of the intestinal tract, mainly consisting of anaerobic bacteria, protects the host against colonization by pathogenic microorganisms. Antimicrobial treatment with ceftriaxone may influence the colonic microflora and as a consequence, the protective effect. Ten healthy volunteers received 1 g of ceftriaxone intramuscularly for five days. This resulted in a significant decrease (p〈0.05) of the mean cultural counts (± SEM) of total anaerobes from 10.67 (0.11) (prior to treatment) to 9.02 (0.45) and 8.97 (0.46) at days 3 and 5, respectively (during treatment). After treatment (days 10 and 15–19), the cultural counts of anaerobes returned to 10.17 (0.16) and 10.44 (0.18), respectively. Bacterial enzymes may serve as an indicator of protective microflora. β- aspartylpeptidase and deoxycholate hydrolase activity was determined in faecal supernatants of the volunteers and compared with anaerobic culturing. Both enzymatic activities show a significant correlation with the total number of anaerobes present at day 3 of ceftriaxone treatment. At day 5 and 8 only β-aspartylpeptidase showed significant correlations with cultural counts of total anaerobes,Bacteroides spp. or bifidobacteria. At day 15 to 19 (ten to 14 days after treatment) β-aspartylpeptidase showed only a significant correlation with the number ofBacteroides spp. This indicates that changes in the indigenous bacterial flora during and shortly after treatment with ceftriaxone can be monitored by determination of β-aspartylpeptidase. Recovery of the intestinal flora is difficult to assess in this manner.
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