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The adhesion of fungi to host cells is an important area of study. Knowledge of the molecular mechanisms involved in these
interactions can be used to devise methods to interfere with them. Similar to many pathogens, loss of fungal adhesion to epithelial
or endothelial cell surfaces results in a marked decrease in virulence when evaluated in both in vivo and in vitro disease
models. This review emphasizes literature from the past year and focuses on the molecular mechanisms by which fungi in the
genera Candida, Cryptococcus, Sporothrix, Pneumocystis, and Aspergillus adhere to epithelial and/or endothelial host surfaces. The methodologies used to conduct these studies are also discussed. 相似文献
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By recording the incubation time needed for initial appearance of the red and blue formazans the reliability of the histochemical method for 3beta-HSD was investigated: 1. Prefixation of small tissue blocks with 1% W/V methanol-free formaldehyde (pH=7.2) for up to 30 min preserved morphological integrity as well as maximal enzyme activity. Moreover, the substantivity of formazans and lipids was enhanced. 2. Commercial available glutaraldehyde (pH=7.2) induced SH groups in the tissue (even at 0.1% W/V for 5 min) thereby enhancing the Nothing dehydrogenase reaction. 3. Preextraction of lipids with acetone for 20 min at -30 degree C caused no loss of activity and was an inevitable step if a reliable activity pattern had to be achieved (e.g. in interstitial cells). 4. No diffusion of enzyme was noticed within 30 min of preincubation in phosphate buffer (0.2 M, pH=7.2) at 20 degree C. 5. By using the double-section incubation method no diffusion of 3beta-HSD or rediffusion of NADH or PMSH could be noticed withn 45 min of incubation, provided that low concentrations of NAD (0.1 mg/ml) and PMS (0.003 mg/ml) were balanced against the concentration of Nitro BT (0.5 mg/ml) or Tetranitro BT (1.0mg/ml). 6. The utlity of different inhibitors of alkaline phosphomonoesterase was tested and discussed. 7. By inhibiting alkaline phosphomonoesterase with 0.1 mM of L-p-bromotetramisole or 16 mM of beta-glycerophosphate, 3beta-HSD was shown to be exclusively NAD-linked. 8. Levamisole was a potent inhibitor of NADH-tetrazolium reductase as well as 3 beta-HSD, but not of NADPH-tetrazolium reductase. 9. 3beta-HSD possess SH groups requisite for the activity as this enzyme was totally inhibited by N-ethyl maleimide. 10. Whether alcohol dehydrogenases may use steroids as substrate is discussed; It is concluded that preextraction (by acetone) and/or the use of an inhibitor of alcohol dehydrogenase (1,10-phenanthroline) has to be performed. 11. Propylene glycol was a poor solvent for all substrates and was itself an excellent substrate for alcohol dehydrogenase. 12. Specifications for the ideal solvent of steroid substrates in the histochemical practice are proposed. DMSO showed to be promising as a steroid solvent (e.g. extraction of formazans was considerably lower as compared to DMF). 13. The utilization of substrates was descending in the following order (using 1 mM and 0.1 ml/ml of either DMF or DMSO): epiandrosterone, methandriol, dehydroepiandrosterone and pregnenolone. 14. If DMSO was used as solvent for pregnenolone (but not for the other substrates tested) an evident increase of activity was recorded as compared to DMF. 相似文献
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Roberto Federico Villa Antonella Gorini Siegfried Hoyer 《Neurochemical research》2009,34(12):2102-2116
The effect of ageing and the relationships between the catalytic properties of enzymes linked to Krebs’ cycle, electron transfer
chain, glutamate and aminoacid metabolism of cerebral cortex, a functional area very sensitive to both age and ischemia, were
studied on mitochondria of adult and aged rats, after complete ischemia of 15 minutes duration. The maximum rate (V
max) of the following enzyme activities: citrate synthase, malate dehydrogenase, succinate dehydrogenase for Krebs’ cycle; NADH-cytochrome
c reductase as total (integrated activity of Complex I–III), rotenone sensitive (Complex I) and cytochrome oxidase (Complex
IV) for electron transfer chain; glutamate dehydrogenase, glutamate–oxaloacetate- and glutamate–pyruvate transaminases for
glutamate metabolism were assayed in non-synaptic, perikaryal mitochondria and in two populations of intra-synaptic mitochondria,
i.e., the light and heavy mitochondrial fraction. The results indicate that in normal, steady-state cerebral cortex, the value
of the same enzyme activity markedly differs according (a) to the different populations of mitochondria, i.e., non-synaptic
or intra-synaptic light and heavy, (b) and respect to ageing. After 15 min of complete ischemia, the enzyme activities of
mitochondria located near the nucleus (perikaryal mitochondria) and in synaptic structures (intra-synaptic mitochondria) of the cerebral tissue were substantially
modified by ischemia. Non-synaptic mitochondria seem to be more affected by ischemia in adult and particularly in aged animals
than the intra-synaptic light and heavy mitochondria. The observed modifications in enzyme activities reflect the metabolic
state of the tissue at each specific experimental condition, as shown by comparative evaluation with respect to the content
of energy-linked metabolites and substrates. The derangements in enzyme activities due to ischemia is greater in aged than
in adult animals and especially the non-synaptic and the intra-synaptic light mitochondria seems to be more affected in aged
animals. These data allow the hypothesis that the observed modifications of catalytic activities in non-synaptic and intra-synaptic
mitochondrial enzyme systems linked to energy metabolism, amino acids and glutamate metabolism are primary responsible for
the physiopathological responses of cerebral tissue to complete cerebral ischemia for 15 min duration during ageing. 相似文献