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71.
Hiroyoshi Hoshi Mikio Kan Wallace L. McKeehan 《In vitro cellular & developmental biology. Plant》1987,23(10):723-732
Summary Hepatocytes were isolated from human fetal liver in order to analyze the direct effects of growth factors and hormones on
human hepatocyte proliferation and function. Mechanical fragmentation and then dissociation of fetal liver tissue with a collagenase/dispase
mixture resulted in high yield and viability of hepatocytes. Hepatocytes were selected in arginine-free, ornithine-supplemented
medium and defined by morphology, albumin production and ornithine uptake into cellular protein. A screen of over twenty growth
factors, hormones, mitogenic agents and crude organ and cell extracts for effect on the stimulation of hepatocyte growth revealed
that EGF, insulin, dexamethasone, and factors concentrated in bovine neural extract and hepatoma cell-conditioned medium supported
attachment, maintenance and growth of hepatocytes on a collagen-coated substratum. The population of cells selected and defined
as differentiated hepatocytes had a proliferative potential of about 4 cumulative population doublings. EGF and insulin synergistically
stimulated DNA synthesis in the absence of other hormones and growth factors. Although neural extracts enhanced hepatocyte
number, no effect on DNA synthesis of neural extracts or purified heparin-binding growth factors from neural extracts could
be demonstrated in the absence or presence of defined hormones, hepatoma-conditioned medium or serum. Hepatoma cell-conditioned
medium had the largest impact on both hepatocyte cell number and DNA synthesis under all conditions. Dialyzed serum protein
(1 mg/ml) at 10 times higher protein concentration had a similar effect to hepatoma cell-conditioned medium (100 μg/ml). The
results suggest that hepatoma cell conditioned medium may be a concentrated and less complicated source than serum for purification
and characterization of additional normal hepatocyte growth factors.
This work was supported by NIH grant DK35310.
Editor’s statement Many investigators have struggled with the special problems associated with culture of differentiated hepatocytes.
In this paper attention is given to the specific growth factor requirements for fetal human hepatocytes. The observation that
factors from hepatoma conditioned medium or neural extracts enhanced the growth of the cells may indicate that additional
growth factors are to be identified that are important in the survival and proliferation of hepatocytes, and may also indicate
that the malignant transformation of these cells may involve the production of autocrine growth stimulators. 相似文献
72.
Different hormonal requirements for androgen-independent growth of normal and tumor epithelial cells from rat prostate 总被引:4,自引:0,他引:4
Wallace L. McKeehan Pamela S. Adams Danna Fast 《In vitro cellular & developmental biology. Plant》1987,23(2):147-152
Summary The proliferation of isolated normal prostate epithelial cells from rat and man is androgen-independent and requires cholera
toxin, insulin, dexamethasone, epidermal growth factor (EGF) and one or more polypeptide factors that are concentrated in
bovine neural tissue. The active agents in the neural tissue extract are heparin-binding polypeptides (prostatropins), the
predominant form of which has a molecular weight of 17400 and an acetylalanine at the aminoterminus. Prostatropins supported
a half-maximal increase in normal prostate epithelial cell number at 50 picomolar. The proliferation of primary and serially-cultured
epithelial cells from androgen-responsive Dunning R3327 rat prostate tumors was also androgen-independent, but exhibited dramatic
alterations in response to hormones that stimulated normal cell proliferation. At low cell density, androgen-independent growth
of isolated tumor-derived epithelial cells was independent on cholera toxin, was stimulated by dexamethasone, required insulin
andeither EGFor prostatropin. The presence of either EGF or prostatropin masked the response to the other factor. In the absence of EGF,
purified prostatropins supported a half-maximal increase in tumor cell number at 7 picomolar. Endogenous production of EGF-like
and prostatropin-like factors or both was suggested by the reduced requirement for EGF and prostatropin at high prostate tumor
cell density. These results suggest that anti-hormonal therapies against prostate tumor growth should be based on intervention
with the activity of insulin (or insulin-like factors) or simultaneous intervention with both EGF and prostatropin (or their
homologues).
This work was supported by NIH grants CA 37589 and HL 33847, and grant 1718 from the Council for Tobacco Research.
Editor’s Statement This paper is the first report of the comparison of the hormone requirements of primary cultures of normal
and tumor prostate epithelial cells from the same system. 相似文献
73.
Experiments were conducted to quantify parasitism of Colorado potato beetle,Leptinotarsa decemlineata (Say), by the egg parasitoid,Edovum puttleri Grissell, on 3 different cultivars of eggplant,Solanum melongena L. Levels of parasitism were higher (P<0.05) on ‘Black Pride’ than on other cultivars. The percentage of egg masses that
were parasitized was 1.2-fold higher (P<0.05) on ‘Black Pride’ than on ‘Harris Special’ and ‘White’. The number of eggs per
mass that were parasitized was 1.3- and 1.4- fold greater (P<0.05) on ‘Black Pride’ than on ‘Harris Special’ and ‘White’,
respectively. The percentage of eggs that were parasitized per mass and percentage of emerged adult parasitoids did not differ
(P>0.05) among cultivars; between 2.1- to 2.6- fold more females than males emerged from eggs on all cultivars during the
growing season.Edovum puttleri suppressed the 2nd generation ofL. decemlineata on ‘Black Pride’ and ‘Harris Special’, but did not suppress populations on ‘White’.
相似文献
74.
The identification and efficiency of arthropod predator and parasites related to natural control of cotton leafworm eggs,Alabama argillacea (Huebner), were studied in a cotton field, Jaboticabal, SP, Brazil. Plants were marked randomly and the eggs found on them were indicated by arrow tapes for predation and parasitism observations. To evaluate and identify the arthropod fauna in the row-meter containing the marked plant, visual countings and collections using D-Vac ® and “beat sheet” were used. The average predation rate during the season, in the presence of 23.0 predators per row-meter, was 50.6% and the parasitism byTrichogrammatoidea annulata was 44.9% totaling 95.5% of egg reduction. In decreasing order of abundance, the arthropod predators found were the antPheidole sp., the spiderChrysso clementinae Petrunkevitch, the hemipteraOrius insidiosus Say andCeratocapsus mariliensis Carvalho & Fontes, and the coccinellidHyperaspis festiva (Mulsant). 相似文献
75.
The postribosomal particle of rabbit liver contains protein-synthesis factors and serum albumin mRNA
As demonstrated by indirect immunoprecipitation and polyacrylamide gel electrophoresis, an 85S particle separated by sucrose density-gradient centrifugation from the postribosomal pellet of rabbit liver, is able to synthesize serum albumin if supplemented with both ribosomal subunits and sources of energy. It is retained on heparin bound to Sepharose 4B, contains translatable mRNA and apparently all protein factors required for translation. This particle may represent a highly organized protein synthesizing machinery, the combination of which with ribosomes results in formation of new protein molecules. 相似文献
76.
Abstract: The higher-molecular-weight elongation factor-1 (EF-1H) of the chick brain was observed to contain three subunits (denominated α, β, and γ), contrary to a previous report that the brain EF-1H consisted of aggregates of low-molecular-weight elongation factor- 1 (EF-1L). Crude EF-1H, obtained from 20-day embryonic brain, was treated with 0.4 M ammonium chloride and 0.1 mM GTP, and EF-1βγ, was obtained using a DEAE-Sephadex column equilibrated in 0.025 mM GTP. Both EF-1β, and EF-1γ, were isolated by means of a DE-52 column equilibrated in 6 M urea and were found to have molecular weights of 2.8 and 4.8 × 104, respectively. EF-1β and EF-1γ were also obtained from young rat and calf brains by the same procedures. The molecular weight of the isolated EF-1α was 5 × 104. It was found that EF-1β stimulated the two EF-1α-dependent reactions, i.e., phenylalanyl-tRNA binding (reaction 1) and polyphenylalanine synthesis (reaction 2), and also stimulated the nucleotide exchange reaction in the EF- 1α-guanine nucleotide binary complex (reaction 3). The degrees of stimulation of reactions 1, 2, and 3 by the addition of EF-1β were 2 to 3 times, about 18 times, and 2 to 3 times as much as with EF-1α alone, respectively. The amino acid compositions of EF-1α -1β, and -1γ and EF-2 were very similar to those of other eukaryotic tissues. Thus the constituents and properties of EFs of the brain were found to be basically similar to those of other tissues of eukaryotes, although EF-1β, and EF-1, had not been reported in the brain. A possible physiological significance of EF-1β during brain development is also discussed. 相似文献
77.
Nayna Vyas-Patel 《Journal of nematology》1983,15(4):594-597
First, second, third, early and late fourth-instar larvae, and pupae of Aedes aegypti were infected with Romanomerrais culicivorax and reared at 20, 25, and 30 C. An increase in the ratio of male to female nematodes was observed with increase in host age at the time of infection at each temperature. The number of pupal and late fourth-instar infections was low, but R. culicivorax continued to develop in adult A. aegypti. Since male nematodes were recovered from both male and female hosts infected as late fourth instars or pupae, the sex of the host did not influence the sex of the nematode. 相似文献
78.
Michael Ready Sandra Bird Gail Rothe J.D. Robertus 《Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression》1983,740(1):19-28
It has been known for some time that pokeweed antiviral protein acts by enzymatically inhibiting protein synthesis on eucaryotic ribosome systems. The site of this action is known to be the ribosome itself. In this paper we show that the pokeweed antiviral protein reaction against ribosomes is a strong function of salt concentrations, where 160 mM K+ and 3 mM Mg2+ retards the reaction, while 20 mM K+ and 2 mM Mg2+ allows maximum reaction rate. It is also shown, however, that an unidentified protein in the postribosomal supernatant solution, together with ATP, allows the ribosome to be attacked even in the presence of high salt. Kinetic analysis of the antiviral protein reaction has been carried out under both sets of conditions, and reveals that the turnover number for the enzyme is about 300–400 mol/mol per min. in each case. The Km for ribosomes is 1 μM in the presence of low salt and 0.2 μM at higher salt in the presence of postribosomal supernatant factors plus ATP. The antiviral protein reaction is also shown to be pH dependent and is controlled by a residue with pKa value of approx. 7.0, apparently a histidine. Stoichiometric reaction of the enzyme with iodoacetamide results in a significant loss of antiribosomal activity. 相似文献
79.
Yuhsi Matuq Pamela S. Adams Nozomu Nishi Hidetaro Yasumitsu John W. Crabb Robert J. Matusik Wallace L. McKeehan 《In vitro cellular & developmental biology. Plant》1989,25(6):581-584
Summary Rat prostate extracts contain an abundant 20–22 kilodalton heparin-binding protein with near identical chromatographic properties,
but only 0.2–1% of the mitogenic activity, of bovine brain heparin-binding growth factor-1 (acidic fibroblast growth factor).
Amino terminal amino acid sequence (met-met-thr-asp-lys-asn-leu-lys-lys-lys-ile-glu-gly-asn-trp-arg-thr-val-tyr-leu-ala-ala-ser-?-val-glu-lys-ile-asn-glu-gly-ser-pro)
and immunochemical analysis revealed that the protein is identical to the androgen-dependent protein “probasin”.
This work was supported in part by NCI grant CA37589 (W. L. M., J. W. C.) and the Medical Research Council of Canada (R. J.
M.). 相似文献
80.
J. Bonde 《BioControl》1989,34(2):275-287
Amblyseius barkeri (Hughes) fed onThrips tabaci (Lind.) at 25°C showed an average duration of 2.2, 0.8 and 3.2 days for the egg, larval and nymphal stages, with mortalities
at 1.0, 1.0 and 3.1%, respectively. Females represented 63% of the population and required multiple matings for optimal fertility.
The oviposition period was 20.3 days and the average oviposition rate 2.3 eggs per day. The intrinsic rate of increase was
0.22 per day. The expected life span was 29.6 days for ♀♀ and 27.4 days for ♂♂.A. barkeri ♂♂ and ♀♀ both consumed 3.3 nymphs of thrips per day (mean value for the feeding stages). The larva does not take up food.
In the absence of thripsA. barkeri was able to consume two-spotted spider mites,Tetranychus urticae (Koch), and their eggs, adult broad mites,Polyphagotarsonemus latus (Banks), and pollen of various plants. Cannibalism was observed when food was lacking. Certain morphological features, egglaying,
mating and predatory behaviour are described.
相似文献