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101.
Perin L. Donnini M. Diomede L. Romano M. Tacconi M. T. Luisetti M. Salmona M. 《Cytotechnology》1991,7(1):25-32
An expression vector for G-CSF, pASLB3-3, was constructed and introduced into Namalwa KJM-1 cells (Hosoi et al., 1988), and cells resistant to 100 nM of methotrexate (MTX) were obtained. Among them, the highest producer, clone SC57, was selected and the productivity of this clone was further characterized. The maximal production of G-CSF was at the most 1.8 g/ml/day using a 25 cm2 tissue culture flask, even though the cell number was above 7×105 cells/ml. The limiting factors at high density were analyzed as the deficiency of nutrients, such as glucose, cysteine and serine, and pH control. The depression of specific G-CSF productivity per cell under the batch culture conditions was overcome by using a perfusion culture system, BiofermenterTM (Sato, 1983) with modifications of nutrients supplementation by a dialysis membrane and/or dissolved oxygen (DO) supplementation by microsilicone fibers. ITPSGF medium was modified to elevate concentrations of amino acids and glucose by 2.0- and 2.5-times, respectively. Under the control of pH at 7.4 and DO at 3 ppm, the specific G-CSF productivity was not depressed even at high cell density (above 1×107 cells/ml), and the amount of G-CSF reached 41 g/ml. These results indicated the possibility of finding the optimum culture conditions for the production of recombinant proteins by Namalwa KJM-1 cells.Abbreviations ABTS
2,2-Azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid
- BSA
Bovine Serum Albumin
- BSA-PBS
Phosphate-buffered Saline without Ca2+ and Mg2+ containing Bovine Serum Albumin
- dhfr
Dihydrofolate Reductase
- DO
Dissolved Oxygen
- G-CSF
Granulocyte Colony-stimulating Factor
- HEPES
4-(2-Hydroxyethyl)-1-piperazineethansulfonic Acid
- IFN
Interferon
- MTX
Methotrexate
- PBS(-)
Phosphate-buffered saline without Ca2+ and Mg2+
- Tween-PBS
Phosphate-buffered saline without Ca2+ and Mg2+ containing 0.05% of Tween 20 相似文献
102.
The hypothesis that chronic competitive antagonism of nicotinic acetylcholine receptors (nAChR), even in the absence of immobilization or paralysis, induces proliferation of the receptor and tolerance to the competitive antagonist was tested. Chronic antagonism of the nAChR was achieved in rats by an infusion of d-tubocurarine (dTC) via subcutaneously placed osmotic pumps. After 2 wk of dTC or saline, the neuromuscular pharmacodynamics and nAChR number were examined. No differences in weight gain or mobility were observed between groups. Chronic dTC infusion at 2 wk resulted in a baseline concentration of 0.41 +/- 0.07 (SE) micrograms/ml, which, if achieved acutely, would cause a depression of the twitch tension to 60% of control twitch height. Moreover the experimental group was able to develop a baseline twitch tension of 50 g, similar to that of controls. Despite the baseline dTC concentration in the experimental group, the effective doses of dTC for twitch depression were similar to those of controls. The plasma dTC concentration required for steady-state twitch inhibition was significantly (P less than 0.05) higher in the experimental group (0.83 +/- 0.04 vs. 0.50 +/- 0.15 micrograms/ml) as were the extrajunctional nAChR (19.76 +/- 1.77 vs. 13.37 +/- 1.82 fmol/mg protein). The diaphragmatic nAChR were unaltered. This study confirms that chronic doses of dTC cause tolerance to its effects and proliferation of nAChR even in the absence of immobilization. The absence of nAChR changes in the diaphragm may be due to the higher margin of safety of the diaphragm for muscle relaxants than for peripheral muscles. Intensive Care Unit patients receiving chronic infusions of dTC to facilitate mechanical ventilation will require increased doses with time. 相似文献
103.
G Teasdale S Galbraith L Murray P Ward D Gentleman M McKean 《BMJ (Clinical research ed.)》1982,285(6356):1695-1697
Deciding which head-injured patients should be transferred to a neurosurgical unit can be difficult. Traditional criteria emphasise the development of deteriorating responsiveness but lead to delayed diagnosis and to avoidable mortality and morbidity. To discover if a more liberal admission policy improved results a study was conducted analysing data collected prospectively from 683 patients who had a traumatic intracranial haematoma evacuated in the Glasgow neurosurgical unit between 1974 and 1980. In the first four years, before the change in policy, mortality was 38% but decreased to 29% afterwards. This reflected a reduction in the proportion of patients who talked after injury but who deteriorated into coma before operation--that is, 31% before the change in policy, 16% afterwards. If the potential benefits of CT scanning in the management of head injuries are to be realised patients must be scanned sooner than in the past. This will usually mean that more patients should go to a neurosurgical unit and that simple criteria for transfer should be established. 相似文献
104.
105.
Ward EJ 《Genetics》1979,92(1):223-230
Chiasma frequency was determined from total chiasma counts, and the distribution of these exchanges was determined by the ratio of proximal to distal chiasmata. No effect of trisomy 10 could be demonstrated. Confirmation was obtained of earlier work showing more proximal and fewer distal chiasmata in K10 plants than in controls. However, diakinesis data failed to confirm the ability of K10 to increase total chiasmata as suggested from metaphase I results. 相似文献
106.
A Comparison of the Genetic Infrastructure of the Ye''Cuana and the Yanomama: A Likelihood Analysis of Genotypic Variation among Populations 总被引:1,自引:1,他引:0 下载免费PDF全文
A general procedure is described for measuring and testing population differences in gametic frequencies. The total dispersion among populations is subdivided in hierarchical fashion. The multiple-locus treatment is simply the sum of the single-locus analyses, provided gametic equilibrium obtains among the loci. In the event that gametic equilibrium does not obtain, correlations among loci need to be dealt with.—The analysis is then used to examine the genetic infrastructure of two Indian tribes from South America, the Ye'cuana (Makiritare) and the Yanomama. From historical evidence, we may identify several "clusters" of villages within each tribe. The demographic and cultural practices affecting village formation and the maintenance of peer integrity are rather different in these tribes, however, and lead us to postulate rather different patterns of genetic variation among villages. Analyses of five codominant two-allele loci, four dominant two-allele loci and two complex loci (with four codominant haplotypes each) demonstrate that Yanomama clusters are more disparate than Ye'cuana clusters, as would have been predicted on sociocultural grounds. 相似文献
107.
The polymorphic HL-A histocompatibility system has been studied in North American black families. The family studies show that haplotype frequencies differ between black and white populations. Seven haplotypes (W28,W5; W28,W17; W28, undefined four; W23,W5; W19,W5; undefined LA,W5; and undefined LA, undefined four) were significantly more frequent in blacks than whites, while haplotypes 1,8 and 3,7 were significantly less frequent. Some of these differences may be accounted for by differences in gene frequencies between the two groups; other differences may be explained by linkage disequilibrium in the white population. No significant linkage disequilibrium between the LA and FOUR loci was found in the black population. 相似文献
108.
A histochemical study has been made on the myotomal musculature of the rainbow trout Salmo gairdneri Richardson. Four main types of fibre can be distinguished on the basis of differences in fibre size, lipid content and succinic dehydrogenase and myofibrillar adenosine triphosphatase activity (ATPase activity). Early histological studies have concluded that in addition to a superficial strip of muscle small diameter 'red' fibres occur throughout the trout myotome. These small diameter fibres occurring in the bulk of the myotome are shown to differ from the superficial fibres with respect to their oxidative metabolism and myofibrillar ATPase. The relevance of this finding to studies on the energetics and swimming efficiency of this species are discussed. 相似文献
109.
110.
Guno Haskå 《Microbial ecology》1975,1(1):234-245
Myxobacteria presumably produce extracellular bacteriolytic enzymes when they are growing in soil. In order to study their ecological significance, adsorption experiments were performed with lytic enzymes produced byMyxococcus virescens in casitone media. Different soils as well as montmorillonite and kaolinite can rapidly adsorb the bacteriolytic but not the proteolytic enzymes. About 1 gm of montmorillonite per liter of cell-free culture solution is enough for the adsorption of 97% of the bacteriolytic enzymes. The adsorption per unit weight is about 100 times greater on montmorillonite than on kaolinite. About 40% of the adsorbed enzymes can be eluted with solutions of high pH or high ionic strength. The only desorbed bacteriolytic enzyme is the alanyl-∈-N-lysine endopeptidase. 相似文献