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51.
Allan Sirsj Kerstin rstrand Bertil K gedal G ran Nylander Anders Gidl f 《Free radical research》1996,25(5):385-391
Microdialysis probes were inserted into the tibialis anterior muscle and into the femoral vein of anaesthetised Sprague-Dawley rats for monitoring of reduced (GSH) and oxidized (GSSG) extracellular glutathione. The dialysates were analysed using HPLC. The levels of GSH and GSSG were high immediately after implantation in the skeletal muscle and declined to steady state levels after 90 minutes into the same range as that found in the venous dialysate. Total ischemia was induced two hours after implantation of the dialysis probe after steady state levels had been reached. The extracellular levels of GSH increased during total ischemia and had doubled at the end of the ischemic period compared to preischemic values. During the following initial 30 minutes of reperfusion the levels increased further to four-fold the preischemic levels. The levels of GSSG also increased (100%) during the initial 30 minutes of reperfusion. The extracellular GSH levels remained elevated for 1 hour of reperfusion, but the GSSG levels returned to preischemic levels. The results indicate that intermittent hypoxia or anoxia in muscle tissue through hypoperfusion or ischemia decreases intracellular GSH stores by leakage, reducing the intracellular antioxidative capacity and increasing the risk for oxidative reperfusion injury upon final normalization of tissue blood supply. 相似文献
52.
Katarina Jansson Gunnar Kratz Anders Haegerstrand 《In vitro cellular & developmental biology. Animal》1996,32(9):534-540
Summary Reepithelialization of artificial partial thickness wounds made in biopsies of human skin was determined after 3, 5, or 7
d of incubation, submerged or elevated to the air-liquid interface. The biopsies were reepithelialized within 5–7 d, with
a more complete epidermal healing in wounds exposed to air. Both types of wounds showed similar time-course in deposition
of basement membrane components, as detected by immunofluorescence labeling. Laminin and collagen type VII were deposited
underneath the migrating tips, whereas collagen type IV was detected after reepithelialization. Markers of terminal differentiation
showed a pattern close to normal in the air-liquid incubated wounds after reepithelialization. Involucrin was detected in
the suprabasal regions of the migrating epidermis and thereafter in the upper half of neo-epidermis in the air-liquid incubated
wound. Filaggrin could not be detected in the submerged wounds at any time during healing, whereas wounds exposed to air showed
a well-differentiated epidermis by Day 7. Tritiated thymidine-incorporation indicated proliferation of epidermal and dermal
cells during reepithelialization and a maintained viability, as shown by cultivation of endothelial- and fibroblast-like cells
obtained from the dermis 7 d after wounding.
Reepithelialization in this humanin vitro model is supported by a matrix close to normal with the possibility of extracellular influences and cell-cell interactions
and, in addition, the technique is simple and reproducible. Therefore, we suggest this model for studies of regeneration in
culture and as a complement toin vivo studies on epidermal healing. 相似文献
53.
Characterization of rapeseed myrosinase-binding protein 总被引:12,自引:0,他引:12
Myrosinase-binding proteins (MBPs) were purified from seeds of Brassica napus L. (oilseed rape). The proteins were characterized with respect to amino-acid composition, peptide sequence and isoelectric points. Gel electrophoresis and Western blotting of protein extracts from mature seeds showed the existence of at least ten proteins reacting with a monoclonal anti-MBP antibody and ranging in molecular size from 110 to 30 kDa. Proteins other than MBP reacting with the anti-MBP antibody were assigned as myrosinase-binding protein-related proteins (MBPRPs). Two MBPRPs were purified by immunoaffinity chromatography and characterized with respect to partial amino-acid sequence. Sequence identities were found between MBP and MBPRP. Western blot analysis of protein extracts from different tissues of B. napus showed that MBPRP is present in the whole plant, whereas MBP mostly occurs in the mature seed. A double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) was used to investigate the occurrence of MBP and MBPRP in developing seeds of some species in the Brassicaceae family.Abbreviations FPLC
fast protein liquid chromatography
- MBP
myrosinase-binding protein
- MBPRP
myrosinase-bindingprotein-telated protein
- PBS
phosphate-buffered saline 相似文献
54.
Abstract: Prostaglandin E2 (PGE2) delivered to the spinal cord produces an increased sensitivity to noxious (hyperalgesia) and innocuous (allodynia) stimuli. The mechanisms that underlie this effect remain unknown, but a PGE2-evoked enhancement of spinal neurotransmitter release may be involved. To address this hypothesis, we examined the effect of PGE2 on CSF concentrations of amino acids and also the modulatory effect of PGE2 on capsaicin-evoked changes of spinal amino acid concentrations using a microdialysis probe placed in the lumbar subarachnoid space. Amino acids were quantified using HPLC with fluorescence detection. Addition of 1 mM, but not 10 or 100 µM, PGE2 to the perfusate for a 10-min period (flow rate, 5 µl/min) evoked an immediate increase (80–100%) in glutamate (Glu), aspartate (Asp), taurine (Tau), glycine (Gly), and γ-aminobutyric acid (GABA) concentrations. Similarly, capsaicin infusion (0.1–10 µM) induced a dose-dependent increase in Glu, Asp, Tau, Gly, GABA, and ethanolamine levels. Significant increases in amino acid levels evoked by PGE2 or capsaicin were associated with a touch-evoked allodynia. The combination of PGE2 (10 µM) and capsaicin (0.1 or 1.0 µM) at concentrations that individually had no effect together evoked a significant increase (60–100%) in Glu, Asp, Tau, Gly, and GABA concentrations and produced tactile allodynia. These data demonstrate that spinally delivered PGE2 or capsaicin substantially elevates CSF concentrations of both excitatory and inhibitory amino acids. The capacity of PGE2 to enhance and prolong capsaicin-evoked amino acid concentrations may be one of the mechanisms by which spinal PGE2 produces hyperalgesia and allodynia. 相似文献
55.
56.
Anders Johansson Gran Widmalm Per-Erik Jansson Stephen G. Wilkinson 《Carbohydrate research》1995,270(2):191-199
The structure of the acidic polysaccharide from Serratia marcescens serogroup O1 has been investigated. NMR spectroscopy together with sugar and methylation analysis have been used as well as a uronic acid degradation. The polysaccharide consists of pentasaccharide repeating units having the following structure.
The polysaccharide also contains one equivalent of O-acetyl groups per repeating unit present on, inter alia, a hydroxymethyl group. 相似文献
57.
Amanda C. J. Vincent Anders Berglund Ingrid Ahnesjö 《Environmental Biology of Fishes》1995,44(4):347-361
Synopsis Synopsis Pipefishes have rarely been watched in the wild and have never before been followed in their common seagrass habitats. This study explores the reproductive ecology of five species of pipefishes living in a Swedish eelgrass meadow during parts of four breeding seasons, tagging four of the species. Pipefish are remarkable for their specialised paternal care: only males aerate, osmoregulate and nourish the developing embryos. Two of the species (Entelurus aequoreus andNerophis ophidion) have simple ventral gluing of eggs on the trunk while three species (Syngnathus acus, S. rostellatus andS. typhle) have fully enclosed brood pouches on their tails. Males of the former species receive eggs from one female while males of the genusSyngnathus receive partial clutches from several females. Sex ratios of adults on the site differed from equal to male-biased to female-biased, according to species.S. typhle were most numerous and were resighted most often. They were present throughout the breeding season whereas there were temporal shifts in the presence of the other species on the meadow and in some sex ratios. Most species occurred in the deeper, denser part of the meadow but there was some habitat separation by species and sex. All species tended to stay low in the eelgrass, primarily coming up above the eelgrass to display and mate. No species showed site fidelity either to a home range or to the meadow, withE. aequoreus adults spending least time on the meadow. Sexual size dimorphism differed: males were larger inS. rostellatus, the same size inS acus and smaller in the other species. Although the species overlap in habitat requirements and breeding season, the only observed interspecific interactions were abortive courtships betweenSyngnathus species. 相似文献
58.
A direct chiral chromatographic reversed phase method for the determination of the enantiomers of felodipine is described. The influence of charged and uncharged modifiers as well as the effect of the mobile phase pH on the enantiomeric resolution is discussed. A high mobile phase pH and the addition of 2-propanol as organic modifier gave the highest separation factor (α = 1.3). The high mobile phase pH (pH = 7.6) is outside the recommended pH limit of silica based columns but was necessary to achieve baseline resolution of (R)- and (S)-felodipine. Improvement of column efficiency by increasing column temperature was utilized for optimization of the enantiomeric resolution (Rs = 1.7). The enantiomers of felodipine and three related compounds were separated within 15 min. The enantiomeric purity of (R)- and (S)-felodipine in injections and (R)-felodipine in bulk substance was higher than 99.5% and no racemization was observed after storage at accelerated conditions. A poor Chiral-AGP® column used for a long period was restored using a simple wash step together with repacking the top of the chromatographic column. © 1995 Wiley-Liss, Inc. 相似文献
59.
Peter Allard Jüri Jarvet Anders Ehrenberg Astrid Gräslund 《Journal of biomolecular NMR》1995,5(2):133-146
Summary The peptide hormone motilin was synthesised with a 13C-enriched -carbon in the leucine at position 10. In aqueous solution, six different relaxation rates were measured for the 13C–H fragment as a function of temperature and with and without the addition of 30% (v/v) of the cosolvent d
2-1,1,1,3,3,3-hexafluoro-2-propanol (HFP). The relaxation rates were analysed employing the spectral density mapping technique introduced by Peng and Wagner [(1992) J. Magn. Reson., 98, 308–322] and using the model-free approach by Lipari and Szabo [(1982) J. Am. Chem. Soc., 104, 4546–4570]. The fit to various models of dynamics was also considered. Different procedures to evaluate the overall rotational correlation time were compared. A single exponential time correlation function was found to give a good fit to the measured spectral densities only for motilin in 30% (v/v) HFP at low temperatures, whereas at high temperatures in this solvent, and in D2O at all temperatures, none of the considered models gave an acceptable fit. A new empirical spectral density function was tested and found to accurately fit the experimental spectral density mapping points. The application of spectral density mapping based on NMR relaxation data for specific 13C–1H vector is shown to be a highly useful method to study biomolecular dynamics. Advantages are high sensitivity, high precision and uniform sampling of the spectral density function over the frequency range.Abbreviations CD
circular dichroism
- NOE
nuclear Overhauser enhancement
- NOESY
two-dimensional NOE spectroscopy
- INEPT
insensitive nuclei enhanced by polarisation transfer
- DANTE
delays alternating with nutations for tailored excitation
- WALTZ-16
wideband, alternating phase, low-power technique for zero residual splitting
- FID
Free induction decay
- ppm
parts per million
- TSPA
3-trimethylsilyl-(3,3,2,2-d)-propionic acid
- HFP
d
2-1,1,1,3,3,3-hexafluoro-2-propanol
- CPMG
Carr-Purcell-Meiboom-Gill
- TFD
time-resolved fluorescence depolarisation
- CSA
chemical shift anisotropy
Partly presented at the symposium Dynamics and Function of Biomolecules, Szeged, Hungary, July 31–August 2, 1993. 相似文献
60.
G V Brown R F Anders R L Coppel R B Saint A F Cowman H D Stahl K R Lingelbach G F Mitchell M P Alpers D J Kemp 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》1984,307(1131):179-187
A library of cDNA clones expressing proteins of the asexual blood stages of a Papua New Guinean isolate of Plasmodium falciparum (isolate FCQ27/PNG (FC27] was constructed in the bacteriophage vector lambda gt11-Amp3. In an in situ colony immunoassay, human serum was used to identify colonies producing natural immunogens. Sera from donors of defined clinical status, or reactive to a defined subset of natural immunogens were used to identify clones of particular interest (for example, clones reacting with convalescent but not with acute serum or clones expressing the isolate specific S-antigen of FC27). Antisera raised by immunizing mice and rabbits with cloned antigens were used to characterize the P. falciparum proteins corresponding to the antigen-positive clones. Nucleotide sequence analysis of an antigen found on the surface of cells infected with ring stage parasites revealed an unusual sequence coding for eight, four and three amino acid repeats rich in acidic amino acids. The discussion centres on the use of cloned antigens as tools for the analysis of the host-protective immune response and selection of candidate vaccine molecules. 相似文献