The phorbol myristate acetate (PMA) stimulated nutrophil respiratory burst has been considered to simply involve the activation of protein kinase C (PKC). However, the PLD activity was also increased by 10‐fold in human neutrophils stimulated with 100 nM PMA. Unexpectedly, U73122, an inhibitor of phospholipase C, was found to significantly inhibit PMA‐stimulated respiratory burst in human neutrophils. U73122 at the concentrations, which were sufficient to inhibit the respiratory burst completely, caused partial inhibition of the PLD activity but no inhibition on PKC translocation and activation, suggesting that PLD activity is also required in PMA‐stimulated respiratory burst. Using 1‐butanol, a PLD substrate, to block phosphatidic acid (PA) generation, the PMA‐stimulated neutrophil respiratory burst was also partially inhibited, further indicating that PLD activation, possibly its hydrolytic product PA and diacylglycerol (DAG), is involved in PMA‐stimulated respiratory burst. Since GF109203X, an inhibitor of PKC that could completely inhibit the respiratory burst in PMA‐stimulated neutrophils, also caused certain suppression of PLD activation, it may suggest that PLD activation in PMA‐stimulated neutrophils might be, to some extent, PKC dependent. To further study whether PLD contributes to the PMA stimulated respiratory burst through itself or its hydrolytic product, 1,2‐dioctanoyl‐sn‐glycerol, an analogue of DAG , was used to prime cells at low concentration, and it reversed the inhibition of PMA‐stimulated respiratory burst by U73122. The results indicate that U73122 may act as an inhibitor of PLD, and PLD activation is required in PMA‐stimulated respiratory burst. 相似文献
Many lung disease processes are characterized by structural and functional heterogeneity that is not directly appreciable with traditional physiological measurements. Experimental methods and lung function modeling to study regional lung function are crucial for better understanding of disease mechanisms and for targeting treatment. Synchrotron radiation offers useful properties to this end: coherence, utilized in phase-contrast imaging, and high flux and a wide energy spectrum which allow the selection of very narrow energy bands of radiation, thus allowing imaging at very specific energies. K-edge subtraction imaging (KES) has thus been developed at synchrotrons for both human and small animal imaging. The unique properties of synchrotron radiation extend X-ray computed tomography (CT) capabilities to quantitatively assess lung morphology, and also to map regional lung ventilation, perfusion, inflammation and biomechanical properties, with microscopic spatial resolution. Four-dimensional imaging, allows the investigation of the dynamics of regional lung functional parameters simultaneously with structural deformation of the lung as a function of time. This review summarizes synchrotron radiation imaging methods and overviews examples of its application in the study of disease mechanisms in preclinical animal models, as well as the potential for clinical translation both through the knowledge gained using these techniques and transfer of imaging technology to laboratory X-ray sources. 相似文献
1. 1.In young pigs living at 35 or 10°C on a high or low energy intake, respiratory enzyme activities in longissimus dorsi muscle were greater both in the cold and on low intake. The elevated activities in the cold were unlikely to be related entirely to shivering since they were also found in muscle from the diaphragm.
2. 2.In a second study, pigs were kept close to thermal neutrality (26°C) on different levels of food intake and for different periods of time. For all animals, as body weight increased there was a decline in respiratory enzyme activity and the number of dark fibres in skeletal muscle. For those of the same weight, but different age and food intake, there was no difference in enzyme activity or number of dark fibres per unit area.
3. 3.At least part of the difference in respiratory enzyme activities related to energy intake must therefore be due to differences in body size. However, size is not the sole determinant of enzyme activity in skeletal muscle, since in animals of similar size those living at 10°C have greater enzyme activities than those at 35°C.
The respiratory system of chemolithoautotrophically-grown Alcaligenes latus contains a, b, and c type cytochromes. Two cytochrome oxidases were identified by their carbon monoxide difference spectra and their differing sensitivities to cyanide and carbon monoxide. The oxidases were cytochrome o and an a-type cytochrome. Ubiquinone was present in A. latus membranes and could be reduced by H2. The quinone analogue, 2-heptyl-4-hydroxy-quinoline-N-oxide (HQNO), was a strong inhibitor of the H2 oxidase reaction, but did not prevent the reduction of either ubiquinone or the cytochromes.Abbreviations HQNO
2-heptyl-4-hydroxy-quinoline-N-oxide
- TMPD
N,N,N,N-tetramethyl-p-phenylenediamine 相似文献
We developed a kinetic assay using a monolayer of differentiated respiratory epithelium in culture to assess bacterial adherence. Mean residence time of bacteria in the tissue culture chamber was estimated from a model-independent (moment) analysis of the rate of bacterial washout from perfused Rose chambers. Results with this method compared favorably with visual assessment of adherence and double radiolabel method with H. influenzae. Adherence was assessed with low inoculae of H. influenzae, P. cepacia and P. aeruginosa avoiding cytotoxic effects seen when large inoculae are added to eukaryotic cells. This method will provide a means of assessing adherence of pathogenic respiratory bacteria to their cellular target at low inoculae. 相似文献
A series of morphologieal and locomotor performance variables was measured in a population of newborn garter snakes to determine whether performance capacity has a significant morphological basis in these animals. Morphological traits measured were body length and mass, number of body and tail vertebrae and numbers of vertebral abnormalities. Locomotor performances included burst and mid-distance speed and distance and time crawled before anti-predator displays were assumed. All performance variables were repeatable in daily replicate trials ( P < 0.001). Individual burst speed, mid-distance speed, and distance crawled were significantly correlated pairwise ( P < 0.01). Most morphological and performance variables had a significant mass dependence (static allometry), although the effects were rather weak ( r 2 < 0.1, except for body length): larger animals performed better and had fewer abnormalities. There were significant associations between some morphological traits and locomotor performance. Morphological factors accounted for 19% of the variation in mid-distance speed and 14% of the variation in antipredator behavior by multiple regression analysis. Canonical correlation of all performance and morphological variables simultaneously accounted for 24% of the observed variation in performance. Numbers of body and tail vertebrae (assayed by scale counts) had an interactive effect on speed of locomotion. 相似文献
We examined the effects of the recombinant human colony stimulating factors GM-CSF and G-CSF, cycloheximide (a protein synthesis inhibitor) and dihydrocytochalasin B (a microfilament disrupting agent) upon FMLP (N-formyl-methionyl-leucylphenylalanine)-stimulated O2− production by neutrophils. We confirmed a time dependent augmentation of O2− production following preincubation of neutrophils either alone or with colony stimulating factors. Furthermore, we found that GM-CSF, but not G-CSF, increased O2− production at some concentrations of the stimulus. Preincubation of neutrophils with cycloheximide in the absence of CSF caused a marked fall in O2−-production that was first evident at 2 hours. The fall in O2−-forming capacity caused by cycloheximide was much less pronounced if dihydrocytochalasin B was also included in the preincubation buffer. These findings suggest a previously unrecognized role for de novo protein synthesis in maintaining the ability of neutrophils to manufacture O2−, and support earlier studies indicating that the cycling of FMLP receptors between the cell membrane and an intracellular compartment is important in determining the magnitude of the respiratory burst in FMLP-stimulated neutrophils. 相似文献