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61.
62.
The effects of TGF1 on cell cycle events in a rat liver derived epithelial cell line (BL9) and in two in vitro transformants of this line were studied by flow cytometry. Using either ethidium bromide staining or the incorporation of bromodeoxyuridine to evaluate DNA synthesis it was shown that TGF1 prevented the entry of G0/G1 phase BL9 cells into S phase. TGF1 did not exert its inhibitory effect(s) on DNA synthesis by the modulation of early events in the cell cycle. The tumorigenic transformed BL9 cell lines gave contrasting responses to the effects of TGF1. DNA synthesis in a BL9 cell line derived by transfection with an active N-ras oncogene was unaffected by TFG1 and thus appeared refractory to its growth controlling effects. On the other hand cells from a BL9 cell line derived by in vitro transformation with activated aflatoxin B1 retained their sensitivity to the effects of TGF1. Thus the loss of the inhibitory effect of TGF1 on DNA synthesis is not obligatory for the malignant transformation of rat liver epithelial cells.Abbreviations TGF1
transforming growth factor 1
- BSA
bovine serum albumin
- FBS
foetal bovine serum
- BrdUrd
bromodeoxyuridine
- PI
propidium iodide
- PBS
phosphate buffered saline 相似文献
63.
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65.
Simulation and optimization of continuous affinity recycle extraction (CARE), a protein purification unit operation based on protein adsorption to solid phase adsorbents, is described in this paper. Rather than packing conventional adsorbent particles in a fixed bed (column), solid/liquid contact is carried out in well-mixed reactors. Continuous operation is achieved by recirculation of the adsorbent particles between two or more contactors. The feasibility of this purification scheme was established with the recovery and isolation of the enzyme beta-galactosidase from E.coli, using the affinity support PABTG/Agarose. A mathematical model describing system performance was developed. The mathematical model was used to optimize several facets of the system design and operation. The base two-stage contractor design was modified by the addition of an intermediate wash stage as well as the incorporation of multiple adsorption stages. These design modifications serve to increase purification, concentration and recovery while utilizing the same amount of adsorbent. The methodology for defining and optimizing objective functions was developed and experimentally validated. Finally, optimum system start-up protocols, minimizing the time required to reach steady-state operation, were developed and experimentally validated. The impact of early introduction of adsorptive purification in a downstream processing sequence, with CARE, was evaluated and is described. Through the early introduction of a highly specific adsorptive step, significant purification is achieved simultaneously with clarification and concentration. In addition, purification performance in CARE was contrasted with that achievable in conventional column chromatography. 相似文献
66.
Dr. Gordon C. Tucker Michel Delarue Suher Zada Jean-Claude Boucaut Jean Paul Thiery 《Cell and tissue research》1988,251(2):457-465
Summary A family of glycoconjugates has recently been shown to share a common carbohydrate epitope recognized by the mouse monoclonal antibody HNK-1. The specificity of HNK-1 was found to be similar to that of another monoclonal antibody, NC-1. These two IgM monoclonal antibodies were raised after immunization of mice with a human T-cell line and avian neural crest-derived ganglia, respectively. The antigens recognized by these antibodies include the myelin-associated glycoprotein, MAG, a glycolipid of defined structure, and a set of molecules involved in cell adhesion. The timing and pattern of appearance of these antigens are distinct. Moreover, the epitope may be absent on an antigen at a given stage or in a given tissue. Therefore, although the molecules able to carry the NC-1/ HNK-1 epitope are numerous and expressed in various tissues, the use of the monoclonal antibodies on tissue sections has proven adequate for following the migration of avian neural crest cells, the major cell lineage recognized by NC-1 and HNK-1 during early embryogenesis. Analogies in several other species have been found on the basis of HNK-1 reactivity. In this study we show that NC-1 and HNK-1 can be used successfully to label migrating neural crest cells in dog, pig and human. On the other hand, the NC-l/HNK-1 epitope was not present on migrating crest cells in amphibians or mice and was found only transiently on the neural crest of rats. 相似文献
67.
Ilhan Olmez M. Cetin Gulovali Glen E. Gordon Robert I. Henkin 《Biological trace element research》1988,17(1):259-270
Although various proteins and some electrolytes have been measured in human saliva, little systematic data about the major
and minor elemental components of this body fluid have been obtained. In order to obtain such data, concentrations of C, Na,
P, Cl, K, Ca, Sc, Cr, Fe, Co, Zn, Se, Br, Rb, Sb, I, and Cs in human parotid saliva were measured by instrumental nuclear
methods. The data obtained confirmed the relative lack of Zn in saliva of patients with hypogeusia (decreased taste acuity)
and suggested that concentrations of Na, Cl, Br, and Ca followed the order: normals > hypogeusia > hyposmia (decreased smell
acuity). To compare concentrations of elements in saliva with those in blood and urine, absolute concentrations were normalized
to that of Na through the use of a concept called an enrichment factor. On this basis, parotid saliva is relatively depleted
in Se, Zn, and Fe and enriched for most other elements relative to blood plasma indicating that the fluid is not simply a
transudate of blood plasma. Using this same technique, saliva composition was found more similar to urine than blood plasma,
being relatively depleted in Se, Cs, and Co, being enriched in I, Br, and Cr and having about the same relative concentrations
of P, Cl, Zn, Fe, Ca, K, and Rb. As the total body concentrations of many of the enriched elements in saliva are extremely
small, their enrichment in saliva suggests special roles for these elements in the oral cavity. Because of its accessibility,
ease of collection, and interaction with some body constituents, saliva represents a useful, albeit neglected, tool in the
diagnosis of some physiological and pathological changes in body function and in understanding important aspects of trace
metal metabolism. 相似文献
68.
Gordon Allen 《Origins of life and evolution of the biosphere》1988,18(3):289-298
Polynucleotides and proteins coupled in mutual synthesis are widely believed to have been needed for the origin of life, but this theory encounters grave problems. Simple catalysts reproducing by positive feedback, sometimes advocated as an alternative, lack a built-in mechanism for generating and accumulating genetic information. Modern organisms, however, integrate genetic information by extrinsic in addition to intrinsic mechanisms, and extrinsic mechanisms were available even at the beginning of chemical evolution for any self-reproducing entities that might have appeared. Novel molecules were generated by reactions among prevailing molecules, and a catalyst multiplying by positive feedback would have transmitted structural information not only to progeny molecules of its kind, but to derivatives and by-products. New molecules derived immediately or remotely from successfully reproducing catalysts would be favored to have catalytic properties. New catalysts with effective positive feedback would increase autocatalytically and be integrated with others into a metabolizing system by natural selection. 相似文献
69.
Missense mutation in the lacI gene of Escherichia coli. Inferences on the structure of the repressor protein 总被引:7,自引:0,他引:7
A J Gordon P A Burns D F Fix F Yatagai F L Allen M J Horsfall J A Halliday J Gray C Bernelot-Moens B W Glickman 《Journal of molecular biology》1988,200(2):239-251
The lac repressor has been studied extensively but a precise three-dimensional structure remains unknown. Studies using mutational data can complement other information and provide insight into protein structure. We have been using the lacI gene-repressor protein system to study the mutational specificity of spontaneous and induced mutation. The sequencing of over 6000 lacI- mutations has revealed 193 missense mutations generating 189 amino acid replacements at 102 different sites within the lac repressor. Replacement sites are not distributed evenly throughout the protein, but are clustered in defined regions. Almost 40% of all sites and over one-half of all substitutions found occur within the amino-terminal 59 amino acid residues, which constitute the DNA-binding domain. The core domain (residues 60 to 360) is less sensitive to amino acid replacement. Here, substitution is found in regions involved in subunit aggregation and at sites surrounding residues that are implicated in sugar-binding. The distribution and nature of missense mutational sites directs attention to particular amino acid residues and residue stretches. 相似文献
70.
S Natarajan E M Gordon E F Sabo J D Godfrey H N Weller J Pluscec M B Rom J Engebrecht D W Cushman J M Deforrest 《Journal of enzyme inhibition》1988,2(2):91-97
The design rationale for a new series of tripeptide derived angiotensin converting enzyme (ACE) inhibitors, which we term "ketomethylureas", is described. Analogs of tripeptide substrates (i.e. N-benzoyl-Phe-Ala-Pro) in which the nitrogen atom of the scissile amide bond and the adjacent asymmetric carbon atom of the penultimate amino acid residue are formally transposed give rise to this novel class of inhibitors. The most potent ketomethylureas inhibit ACE with I50 values in the nM range. 相似文献