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181.
Further Studies on the Lyo and Desmo Components of Several Hydrolytic Enzymes and Their Histochemical Significance 总被引:1,自引:1,他引:0 下载免费PDF全文
This report describes additional studies of the lyo and desmo components of esterase, alkaline phosphatase, acid phosphatase, leucine aminopeptidase, and β-glucuronidase. The techniques used have already been reported (7). Enzyme diffusion occurs to different degrees in different fixatives, and varies somewhat with each enzyme. Loss of enzymatic activity during fixation occurs as a result of both inactivation due to the chemical reaction of the fixative with the enzymic protein, and diffusion of the lyo component into the fixative. The amount of diffusion into formalin can be reduced by the addition of salts, sucrose, or methocel. The pH of the aqueous medium significantly influences the removal of the lyo fraction from the tissue section. A striking similarity can be noted in the proportions of each fraction of enzyme present in the kidney of the rat, dog, and man. The procedure of fixation and paraffin embedding of tissue blocks does not wholly prevent the diffusion of the lyo component from the tissue sections when they are subsequently immersed in the aqueous incubation medium. 相似文献
182.
A dialysis unit was used to test whether direct physical contact between serum albumin and hamster spermatozoa is required for capacitation and/or the acrosome reaction. Sperm and bovine serum albumin (BSA) were incubated cither together (direct incubation) or separated by a dialysis membrane (indirect incubation). Sperm viability was supported with “sperm motility factors” (hypotaurine and epinephrine) and polyvinylalcohol (PVA). Spermatozoa became capacitated and underwent acrosome reactions when directly incubated in medium containing BSA (TALP-PVA), but did not undergo acrosome reactions when indirectly incubated with BSA (medium TLP-PVA). When sperm were first incubated for 4 hr indirectly with BSA, followed by 4 hr direct incubation with BSA, capacitation did not occur during indirect incubation. These findings indicate that an “intimate association” is necessary between serum albumin and spermatozoa to support capacitation under in vitro incubation conditions. The data are consistent with the concept of direct transfer of compounds from sperm to albumin and/or vice versa during sperm capacitation. 相似文献
183.
Carol L. Williams Vanda A. Lennon Mark R. Pittelkow 《In vitro cellular & developmental biology. Plant》1989,25(5):397-401
Summary A time-dependent redistribution of microfilaments was observed in cultured human keratinocytes using a human monoclonal autoantibody
specific for myosin. Immunofluorescent staining revealed that 5 days after plating keratinocytes in either 0.1 mM or 2.0 mM
Ca++, myosin was distributed uniformly throughout the cytoplasm. At day 6, parallel arrays of myosin-containing microfilaments
were prominent in the cell peripheries. At day 7 the microfilaments formed circumferential rings. The distribution of the
microfilaments was disrupted by cytochalasin but not by colchicine, indicating that this novel distribution of myosin was
not dependent on colchicine-sensitive vimentin intermediate filaments. The time-dependent redistribution of myosin was not
influenced by cell population density, cell shape or cell cycle phase, except for mitotic cells in which myosin was distributed
diffusely through the cytoplasm. If, as suggested by Kolega (9), microfilaments align parallel to the direction of applied
tension, the redistribution of myosin-containing microfilaments in cultured keratinocytes may reflect the increased tension
between cells resulting from increasing strength of cell-cell junctions over time. In sectioned human skin, myosin was localized
in the peripheral cytoplasm of stratified epidermal cells. Tensions arising from the numerous desmosomal junctions between
cellsin vivo could account for this distribution of myosin.
Supported by grant NS-23537 (V. A. L.) from the National Institutes of Health, Bethesda, MD, and by the Mayo Foundation. C.
L. W. is recipient of the Kermit E. Osserman and Blanche McClure Fellowship, 1987, National Myasthenia Gravis Foundation. 相似文献
184.
We quantified microscale pattern in vegetation and seed assemblages along a 24 m transect before and for two years following a controlled burn in chamise (Adenostoma fasciculatum) chaparral in central coastal California. Our objective was to document scale-dependent correlation between pre-burn seed assemblages, pre-burn canopy cover, microtopography, soil temperatures during burning, post-burn seed assemblages and post-burn vegetation. Scale-dependent correlations among pre- and post-burn seed densities, maximum soil temperatures during burning, microtopography and post-burn vegetation were measured based on two-term local covariance analysis.Seed distribution varied among species prior to fire, with seeds of some annual species concentrated in gap areas. Maximum soil temperatures during burning ranged from less than 50 °C to 225 °C, and were generally lowest in gaps in the pre-burn canopy. These gaps were associated with local topographic depressions. After burning, readily germinable seeds were concentrated in or near gaps in the pre-burn canopy. Germination of different species was variously enhanced, diminished or unchanged by the passage of fire. Post-burn vegetation was very patchy, with some areas nearly devoid of seedlings and other areas, especially pre-burn canopy gaps, supporting numerous seedlings. Seedling recruitment patterns in the second year were generally highly correlated with patterns in pre-burn seed banks and first year vegetation. Although many species exhibited similar recruitment patterns, several different mechanisms may have been responsible for the origin of those patterns. 相似文献
185.
Summary The ages of 252 mature female krill, collected from Prydz Bay in January 1985, were determined using length frequency analysis and the fluorescent age pigment (FAP) technique. Results of both methods suggest 6 years classes for adult krill. Correspondence between the ages determined by the two techniques is generally within one year. The animals were also analyzed by a computerized image analysis system, which recorded a large suite of size and shape parameters. The accuracy of discriminant functions constructed to relate the image analysis parameters to age approached 90% for the ages defined by length frequency, and 52% for physiological age. 相似文献
186.
Pretreatment of rat brain membranes at pH 4.5 before assay at pH 7.4 modifies the function of GTP-binding proteins (G-proteins) by eliminating Gs-stimulated adenylate cyclase activity while increasing opiate-inhibited adenylate cyclase activity. To help characterize the molecular nature of the low pH effect, we labeled Gs and Gi alpha-subunits in both control and low pH-pretreated membranes with the GTP photoaffinity analog [32P]P3 (4-azidoanilido)-P1-5'-GTP ([32P]AAGTP). When membranes were preincubated with unlabeled AAGTP, a persistent inhibitory state of adenylate cyclase was produced, which was overcome in untreated membranes with high (greater than 1 microM) concentrations of guanylyl-5'-imidodiphosphate [Gpp(NH)p]. In low pH-pretreated membranes, this inhibition could not be overcome, and stimulation by Gpp(NH)p was eliminated. Maximal inhibition of adenylate cyclase achieved by incubation with AAGTP was not altered by low pH pretreatment. Incorporation of [32P]AAGTP into Gs (42 kilodaltons) or Gi/o (40 kilodaltons) was unaffected by low pH pretreatment; however, transfer of 32P from Gi/o to Gs, which occurs with low (10 nM) concentrations of Gpp(NH)p in untreated membranes, was severely retarded in low pH-pretreated membranes. Both the potency and efficacy of Gpp(NH)p in producing exchange of [32P]AAGTP from Gi/o to Gs were markedly reduced by low pH pretreatment. These results correlate the loss of Gs-stimulated adenylate cyclase with a loss of transfer of nucleotide from Gi/o to Gs alpha-subunits and suggest that the nucleotide exchange participates in the modulation of neuronal adenylate cyclase. 相似文献
187.
In a spheroplasting method which allows the fractionation and quantification of cloned invertase activity in recombinantSaccharomyces cerevisiae cells, the yeast cell is selectively degraded with the enzyme Zymolyase for 60 minutes at 45°C to separate periplasmic proteins from cytoplasmic proteins. Most of the glucose-6-phosphate dehydrogenase (a cytoplasmic marker protein) was found in the cytoplasmic fraction. 相似文献
188.
Sadettin S. Ozturk Mark E. Meyerhoff Bernhard O. Palsson 《Biotechnology Techniques》1989,3(4):217-222
Summary This paper describes the use of a commercially available off-line gas sensing electrode for determination of ammonia and glutamine in cell culture media. The measurement technique was tested in different media preparations with different serum concentrations. The glutamine decomposition was studied as a function of pH for cell culture medium and the results were compared to those obtained by conventional methods,i.e., HPLC. Finally, glutamine and ammonia metabolism were studied during the cultivation of a hybridoma cell line. 相似文献
189.
Howard Griffiths Mark S. J. Broadmeadow Anne M. Borland Clive S. Hetherington 《Planta》1990,181(4):604-610
Short-term measurements of instantaneous carbon-isotope discrimination have been determined from mass-spectrometric analyses of CO2 collected online during gas exchange for the epiphytic bromeliad Tillandsia utriculata L. Using this technique, the isotopic signature of CO2 exchange for each phase of Crassulacean acid metabolism (CAM) has been characterised. During night-time fixation of CO2 (Phase I), discrimination () ranged from 4.4 to 6.6, equivalent to an effective carbon-isotope ratio (13C) of –12.3 to –14.5 versus Pee Dee Belemnite (PDB). These values reflected the gross photosynthetic balance between net CO2 uptake and refixation of respiratory CO2, characteristic of CAM in the Bromeliaceae. When for the relative proportion of external (p
a
) and internal (p
i) CO2 is taken into account, calculated p
i/p
a decreased during the later part of the dark period from 0.68 to 0.48. Measurements of during Phase II, early in the light period, showed the transition between C4 and C3 pathways, with carboxylation being increasingly dominated by ribulose bisphosphate carboxylase (Rubisco) as increased from 10.5 to 21.2 During decarboxylation in the light period (Phase III), CO2 leaked out of the leaf and the inherent discrimination of Rubisco was expressed. The value of calculated from on-line measurements (64.4) showed that the CO2 lost was considerably enriched in 13C, and this was confirmed by direct analysis of the CO2 diffusing out into a CO2-free atmosphere (
13C = + 51.6 versus PDB). Instantaneous discrimination was characteristic of the C3 pathway during Phase IV (late in the light period), but a reduction in showed an increasing contribution from phosphoenolpyruvate carboxylase. The results from this non-invasive technique confirm the observations that double carboxylation involving both phosphoenolpyruvate carboxylase and Rubisco occurs during the transient phases of CAM (II and IV) in the light period.Abbreviations and Symbols CAM
Crassulacean acid metabolism
-
H+
(dawn-dusk) variation in titratable acidity
-
13C
carbonisotope ratio of plant organic material, relative to Pee Dee Belemnite (vs. PDB)
-
discrimination against 13CO2,
-
p
i, p
a
internal, external partial pressures of CO2
- Rubisco
ribulose1,5-bisphosphate carboxylase
- PAR
photosynthetically active radiation
- PEPCase
phosphoenolpyruvate carboxylase
We are grateful for financial support in respect of research grants (GR3/5360, GR3/6419) and a studentship awarded by the Natural Environment Research Council, UK. 相似文献
190.