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1.
2.
Expression of tracheal differentiated functions in serum-free hormone-supplemented medium 总被引:18,自引:0,他引:18
Most dissociated airway epithelial cells in culture express few of their in vivo functions and only to a limited degree. In this report, we demonstrate that hamster tracheal epithelial (HTE) cells cultured on a collagen gel substratum in a serum-free hormone-supplemented medium differentiate to cilia-beating and mucus-secreting cell types. The medium is Ham's F-12 supplemented with insulin, epidermal growth factor, transferrin, hydrocortisone, cholera toxin, bovine hypothalamus extract, and vitamin A. Under these culture conditions, HTE cells exhibit a growth rate of 24 h/population doubling and reach confluency, at a density of 2-5 X 10(4) cells/cm2, within 2 weeks. Both the collagen gel substratum and vitamin A of this culture system are important to the growth and differentiation of HTE cells in vitro. Evidence of HTE cell differentiation has been obtained at both the ultrastructural and the histochemical levels. In addition, a variety of biochemical studies (gel filtration, ion exchange column chromatography, enzyme digestion, nitrous acid treatment, and composition analysis) indicate the production of mucin-like glycoprotein in the HTE cultures. The levels of mucin-like glycoprotein were found to closely correlate with the histochemically quantitated levels of the mucous cell type. Kinetic studies demonstrate that HTE cells rapidly lose their differentiated features during the attachment stage of primary culture but redifferentiation occurs after the cultures reach confluency. The ability of HTE cells to grow and differentiate in this serum-free culture system in the absence of other cell types should greatly facilitate the study of mucociliary functions in vitro. 相似文献
3.
4.
Folylpolyglutamates in Leishmania major 总被引:3,自引:0,他引:3
D V Santi P Nolan B Shane 《Biochemical and biophysical research communications》1987,146(3):1089-1092
The intracellular folates of the protozoan parasite Leishmania major have been examined. About 95% of the exogenous [3H]folate accumulated by the protozoan is metabolized to polyglutamate conjugates within 65 hr, and the intracellular folates are about forty-fold concentrated over the folate in the medium. The predominant metabolite of folic acid is the pentaglutamate conjugate (85%), with lessor amounts of the tetraglutamate (approximately 9%) and hexaglutamate (approximately 3%), and trace (less than 2.5%) amounts of di-, tri- and hepta-glutamate conjugates. Chromatographic properties of the products indicate that the conjugates are linked through the gamma-carboxyl groups. The folylpolyglutamate distribution in Leishmania is similar to that found in mammalian tissues. 相似文献
5.
6.
Valérie Toulon Hervé Sentenac Jean-Baptiste Thibaud André Soler David Clarkson Claude Grignon 《Planta》1989,179(2):235-241
The effect of HCO
3
-
on ion absorption by young corn roots was studied in conditions allowing the independent control of both the pH of uptake solution and the CO2 partial pressure in air bubbled through the solution. The surface pH shift in the vicinity of the outer surface of the plasmalemma induced by active H+ excretion was estimated using the initial uptake rate of acetic acid as a pH probe (Sentenac and Grignon (1987) Plant Physiol. 84, 1367). Acetic acid and orthophosphate uptake rates and NO
3
-
accumulation were slowed down, while 86Rb+ uptake and K+ accumulation rates were increased by HCO
3
-
. These effects were similar to those induced by 4-(2-hydroxyethyl)-1-piperazineethane sulfonic acid/2-amino-2-(hydroxymethyl)-1,3-propanediol (Hepes-Tris). They were more pronounced when the H+ excretion was strong, were rapidly reversible and were not additive to those of Hepes-Tris. The hypothesis is advanced that the buffering system CO2/H2CO3/HCO
3
-
accelerated the diffusion of equivalent H+ inside the cell wall towards the medium. This attenuated the surface pH shift in the vicinity the plasma membrane and affected the coupling between the proton pump and cotransport systems.Abbreviations FW
fresh weight
- Hepes
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- Jaa
acetic acid influx
- JK
+
K+ influx
- JPi
orthophosphate influx
- Mes
2-(N-morpholino)ethanesulfonic acid
- pCO2
CO2 partial pressure
- Tris
2-amino-2-(hydroxymethyl)-1,3-propanediol 相似文献
7.
A firm, mobile, oblong mass was discovered as an incidental finding in the stomach of a chimpanzee (Pan troglodytes) during routine surgery. It was removed and determined to be a trichobezoar. 相似文献
8.
Summary Fertile transgenic plants of the annual pasture legume Medicago truncatula were obtained by Agrobacterium-mediated transformation, utilising a disarmed Ti plasmid and a binary vector containing the kanamycin resistance gene under the control of the cauliflower mosaic virus 35S promoter. Factors contributing to the result included an improved plant regeneration protocol and the use of explants from a plant identified as possessing high regeneration capability from tissue culture. Genes present on the T-DNA of the Ri plasmid had a negative effect on somatic embryogenesis. Only tissue inoculated with Agrobacterium strains containing a disarmed Ti plasmid lacking the T-DNA region or a Ri plasmid with an inactivated rol A gene regenerated transgenic plants. Fertile transgenic plants were only obtained with disarmed A. tumefaciens, and the introduced NPT II gene was transmitted to R1 progeny.Abbreviations BAP
6-benzylaminopurine
- NAA
1-naphthaleneacetic acid
- NPT
neomycin phosphotransferase 相似文献
9.
DNA binding and I kappa B inhibition of the cloned p65 subunit of NF-kappa B, a rel-related polypeptide 总被引:140,自引:0,他引:140
The sequence and biochemical properties of the product of the cloned cDNA for the p65 subunit of nuclear factor kappa B (NF-kappa B) have been determined. The cDNA has an open reading frame of 549 amino acids capable of encoding a 60 kd protein. NF-kappa B p65 contains an amino-terminal region of 320 amino acids with extensive similarity to the oncogene c-rel and lesser similarity to NF-kappa B p50. In vitro translated p65 forms a DNA-binding complex with NF-kappa B p50, and the binding of this complex can be specifically inhibited by purified I kappa B. Progressive carboxy-terminal deletions of p65 show that, contrary to previous assumptions, p65 does include a DNA-binding domain that in vivo might become activated only through hetero-oligomerization with p50. DNA binding by truncated p65 is inhibited by I kappa B, thus mapping the I kappa B interaction domain to the rel-homologous region and suggesting that I kappa B exerts its inhibitory effect upon NF-kappa B primarily through interaction with p65. 相似文献
10.
Valérie Maxime Jean-Pierre Pennec Claude Peyraud 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1991,161(6):557-568
Summary The effects of increased ambient salinity (35 mg · ml-1) were studied at 1, 6, and 24 h after direct transfer of rainbow trout from freshwater to seawater. Two series of experiments were carried out successively. The first series was designed to simultaneously study all the respiratory (except Hb affinity for O2), circulatory, and acid-base variables in each fish. In this series, fish were fitted with catheters chronically inserted into the cardiac bulbus, the dorsal aorta, and the opercular and buccal cavities. In the second series, designed to study haemoglobin O2 affinity, fish were fitted with only a dorsal aorta catheter. The ventilatory flow (
) was markedly increased just after transfer (by 55% at 1 h), then more moderately (by 20% at 6 h and 32% at 24 h). The initial hyperventilation peak was associated with frequent couphing motions. These ventilatory changes resulted essentially from increase in ventilatory amplitude. Initially, standard oxygen consumption (MM}O2) decreased slightly, the moderately increased (by 12% at 24 h), so that the oxygen convection requirement (
) increased substantially. In spite of an increased ventilation, the partial pressure of oxygen in arterial blood (P
aO2) decreased slightly at 1 h, prior to returning to control levels, while partial pressure of carbon dioxide in arterial blood (P
aCO2) was not significantly decreased. Gill oxygen transfer factor decreased substantially at 1 h (by 35%) then more moderately (by 7% at 1 h and 12% at 24 h). These results suggest a decrease in gas diffusing capacity of the gills. As P
aCO2 remained approximatively unchanged, the gradual decrease in arterial pH (pHa) from 7.94 to 7.67 at 24 h must therefore be regarded as a metabolic acidosis. The strong ion difference decreased markedly because the concentration of plasma chloride increased more than that of sodium. Arterial O2 content (C
aO2) gradually decreased (by 38% at 24 h) simultaneously with the decrease in pHa, while the ratio P
aO2/C
aO2 increased. In parallel, seawater exposure induced a marked decrease in affinity of haemoglobin for O2, so that at 24 h, P50 was increased by 26% above the value obtained in freshwater-adapted trout. The increase in
could be ascribed initially (at 1 h) to the decrease of P
aO2 and later to a stimulation of respiratory neurons resulting from the lowered medullary interstitial pH. The decrease in C
aO2 could be interpreted mainly as a consequence of a decreased affinity of haemoglobin for O2, likely to be due to the blood acidosis and a predictable increase in chloride concentration within erythrocytes. Cardiac output (
) slightly decreased at 1 h, then progressively increased by 30% at 24 h. Branchial vascular resistance increased at 1 h by 28%, then decreased by 18% of the control value at 24 h. Systemic vascular resistance decreased markedly by 40% at 24 h. As heart rate (HR) remained significantly unchanged, the cardiac stroke volume initially decreased then increased in relation to the changes in
. The increase of
, allowing compensation for the effect of decreased C
aO2 in tissue O2 supply, was interpreted as a passive consequence of the decrease in total vascular resistance occurring during seawater exposure.Abbreviations a.u.
arbitrary units
-
C
aO2
arterial oxygen content
- pH50
arterial pH at P50
-
C
vO2
venous oxygen content
- Hb
haemoglobin
- HR
heart rate
- Hct
hematocrit
- nHill
Hill coefficient
- O2
standard oxygen consumption
-
P
aCO2
arterial partial pressure of carbon dioxide
-
P
aO2
arterial partial pressure of oxygen
-
P
vO2
oxygen partial pressure in mixed venous blood
- P50
oxygen tension at half saturation of haemoglobin
-
P
VA, P
DA
blood pressure in ventral and dorsal aorta
- pHa
arterial pH
-
PIO2, PEO2
oxygen partial pressure of inspired and expired water
-
PO2
oxygen partial pressure
-
cardiac output
- SEM
standard error of mean
- S.I.D.
strong ion difference
- SV
cardiac stroke volume
- TO2
gill oxygen transfer factor
- U
oxygen extraction coefficient
- VA
ventilatory amplitude
- VF
ventilatory frequency
- VRG, VRS
branchial and systemic vascular resistances
-
ventilatory flow
-
ventilatory oxygen convection requirement 相似文献