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221.
Roberto E. Izquierdo Kimberly Breese Shalini Jain Daniel Carestio Lawrence Jung James Figge 《In vitro cellular & developmental biology. Animal》1995,31(1):71-76
Summary Gene transfer techniques can be used to encode the production of a polypeptide product, such as human growth hormone (hGH),
that is missing in an acquired or inherited disease state such as growth hormone deficiency. In one model system, engineered
C2C12 myoblasts are injected intramuscularly into a mouse and subsequently secrete hGH into the circulation. In this regard,
a gene-expression regulatory system that functions in myoblasts would be of interest. We demonstrate that theEscherichia coli lac operon system can be used to stringently regulate the expression of hGH in engineered C2C12 myoblasts in tissue culture.
A DNA segment encoding hGH was linked to a DNA segment containing an SV40 enhancer and promoter. The latter components were
positioned between two syntheticlac operators.Lac repressor expression was driven by a simian cytomegalovirus promoter. In transient co-transfection assays, hGH expression
from cultured C2C12 myoblasts could be modulated up to 60-fold (P = 0.002) with the inducing agent, isopropyl-β-d-thiogalactoside (IPTG). In the absence of IPTG, hGH expression was almost fully repressed. These results show that the components
of theE. coli lac operon provide a stringent regulatory system for use in myoblasts. The system might prove to be useful for the regulation
of transferred genes in animals. 相似文献
222.
Andrew F. Rowley John Knight Paul Lloyd-Evans Jason W. Holland Philip J. Vickers 《Fish & shellfish immunology》1995,5(8)
Eicosanoids have been demonstrated to play a central role in immune regulation in mammals brought about by their direct effects on cells such as macrophages and lymphocytes or by their indirect effects via cytokines. Studies have shown that fish mononuclear phagocytes, granulocytes and thrombocytes synthesize and release both cyclooxygenase- and lipoxygenase-derived products such as prostaglandin E2, leukotriene B4 and lipoxin A4. Whether lymphocytes have the ability to generate leukotrienes and lipoxins is still unclear but they do appear to have 12-lipoxygenase activity that leads to the generation of 12-hydroxy fatty acid derivatives. As in mammals, leukotriene and lipoxin biosynthesis requires the presence of a 5-lipoxygenase activating protein-like molecule that is sensitive to the action of the specific inhibitor, MK-886. The prostaglandin-generating ability of trout macrophages can be altered by incubation with lipopolysaccharide suggesting the possible presence of an inducible cyclooxygenase activity. Prostaglandins have been found to suppress the mitogen-induced proliferation of trout leucocytes and the generation of humoral antibody and plasma cells both in vivo and in vitro. The lipoxygenase products, leukotriene B4 and lipoxin A4 have more variable effects ranging from inhibition to stimulation depending on the assay system employed. Overall, there is clear evidence that eicosanoids play a role in immune regulation in fish in a similar way to that reported in mammals. 相似文献
223.
S. M. Thoroed K. Fugelli 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1994,164(1):1-10
Erythrocytes from the marine fish species ballan wrasse (Labrus berggylta Ascanius), bullhead (Myoxocephalus scorpius L.), cod (Gadus morhua L.), dab (Limanda limanda L.), eelpout (Zoarces viviparus L.), flounder (Platichthys flesus L.), lumpfish (Cyclopterus lumpus L.), plaice (Pleuronectes platessa L.), sole (Solea solea L.) and turbot (Scophthalmus maxima L.) possess the capacity for regulatory volume decrease. This property was demonstrated in vitro by reduction of the osmolality of the incubation medium from 330 to 255 mosmol·kg-1. During the 4-h incubation period only the lumpfish cells completely regained the original volume. Twenty-seven free amino compounds were present in detectable amounts in the erythrocytes. At normal osmolality the taurine content was between 14.0 mol·g dry weight-1 (lumpfish) and 147.4 mol·g dry weight-1 (sole). Except in the bullhead, taurine was the quantitatively dominating amino compound in the erythrocytes from all species, and accounted for betwee 23% (lumpfish) and 88% (sole) of the total content of free amino compounds. In each species the regulatory volume decrease was associated with a reduction in the cellular content of taurine. Taurine contributed to between 6% (lumpfish) and 36% (flounder) of the cell shrinkage. There was a significant negative correlation, however, between the cellular concentration of taurine at normal osmolality and the capacity of the cells for regulatory volume decrease. Gamma-aminobutyric acid and/or glycine also contributed to the process of volume regulation, but to a lesser extent than taurine. The volume regulatory efflux of taurine and -aminobutyric acid were mediated by taurine channels. It is suggested that these channels also mediated the reduction in the cellular contents of glycine.Abbreviations cmp
counts per minute
- dw
dry weight
- GABA
-amino-n-butyric acid
- MW
molecular weight
- SD
standard deviation
- ww
wet weight 相似文献
224.
Epithelial cells isolated from rat lung and trachea were grown on monolayers and their response to a number of hormones and
growth factors were studied. Maximum proliferative response in serum containing media was observed when insulin, cholera toxin
and cortisol were present together. However, these additives when present independently showed a marginal response. The synergism,
due to these factors in promoting growth was seen very early in culture (day 4) as shown by thymidine labelling studies, On
examining the indices of early mitogenesis, such as the expression ofc-myc, our data suggests that these factors stimulate the expression ofc-myc within 4 h. With respect to expression of TNF-α mRNA, this study suggests a possible modulation of TNF-α expression in response
to these mitogens that stimulate proliferation maximally. Whether this expression of TNF-α by these epithelial cells is due
to a maximal proliferative stimulus and/or is an early step in the cascade of intracellular signalling events is to be investigated
in detail. 相似文献
225.
Saks V. A. Khuchua Z. A. Vasilyeva E. V. Belikova O. Yu Kuznetsov A. V. 《Molecular and cellular biochemistry》1994,133(1):155-192
The published experimental data and existing concepts of cellular regulation of respiration are analyzed. Conventional, simplified considerations of regulatory mechanism by cytoplasmic ADP according to Michaelis-Menten kinetics or by derived parameters such as phosphate potential etc. do not explain relationships between oxygen consumption, workload and metabolic state of the cell. On the other hand, there are abundant data in literature showing microheterogeneity of cytoplasmic space in muscle cells, in particular with respect to ATP (and ADP) due to the structural organization of cell interior, existence of multienzyme complexes and structured water phase. Also very recent experimental data show that the intracellular diffusion of ADP is retarded in cardiomyocytes because of very low permeability of the mitochondrial outer membrane for adenine nucleotidesin vivo. Most probably, permeability of the outer mitochondrial membrane porin channels is controlled in the cellsin vivo by some intracellular factors which may be connected to cytoskeleton and lost during mitochondrial isolation. All these numerous data show convincingly that cellular metabolism cannot be understood if cell interior is considered as homogenous solution, and it is necessary to use the theories of organized metabolic systems and substrate-product channelling in multienzyme systems to understand metabolic regulation of respiration. One of these systems is the creatine kinase system, which channels high energy phosphates from mitochondria to sites of energy utilization. It is proposed that in muscle cells feed-back signal between contraction and mitochondrial respiration may be conducted by metabolic wave (propagation of oscillations of local concentration of ADP and creatine) through cytoplasmic equilibrium creatine and adenylate kinases and is amplified by coupled creatine kinase reaction in mitochondria. Mitochondrial creatine kinase has experimentally been shown to be a powerful amplifier of regulatory action of weak ADP fluxes due to its coupling to adenine nucleotide translocase. This phenomenon is also carefully analyzed.It is easier to explain biochemistry in terms of transport than it is to explain transport in terms of biochemistry. P. Mitchell The Ninth Sir Hans Krebs Lecture, Dresden, July 2, 1978. 相似文献
226.
227.
C. S. Degoute C. Dubreuil M. J. Ray J. Guitton M. Manchon V. Banssillon J. L. Saumet 《European journal of applied physiology and occupational physiology》1994,69(5):414-420
Studies by laser-Doppler flowmetry of middle ear microcirculation changes induced by physical and chemical stimuli in the animal have only recently been made. This prospective study, performed in humans, was designed to compare the effects of a postural manoeuvre (headup tilt 30°), hypotension and locally applied vasoconstriction on middle ear blood flow during anaesthesia. Circulatory changes provoked by a headup tilt of 30°, and successive intravenous boluses of potent vasodilators, were compared with circulatory changes provoked by locally applied adrenaline, in ten healthy patients in good physical states undergoing middle ear surgical repair. Heart rate and direct arterial pressure were continuously recorded via a radial artery cannula. Middle ear blood flow was continuously recorded via a laser-Doppler probe placed on the promontorium cavi tympani. Metabolic parameters (partial pressure of O2 and CO2 in arterial blood, pH, arterial lactate concentrations) and arterial concentrations of propofol were measured just before and just after the experiment. Headup tilt did not modify heart rate, mean arterial pressure or middle ear blood flow. Vasodilators (nicardipine, nitroprusside, nitroglycerin) provoked a fall in arterial pressure (P<0.0001,P<0.0001,P<0.019, respectively), but did not induce any significant variations in heart rate; variations occurred in middle ear blood flow (P>0.05, not significant) which were different according to patients and agents. Locally applied adrenaline provoked a fall in the middle ear blood flow (P<0.0012), with no effect on heart rate and arterial pressure. There were no significant changes in metabolic values, or propofol serum concentrations. The behaviour of the middle ear blood flow submitted to hypotension, posture, or to vasoconstriction could be related to counteracting regulatory responses and/or to direct vascular effects. 相似文献
228.
Enrique Cerdá-Olmedo Rafael Fernández-Martín Javier Ávalos 《Antonie van Leeuwenhoek》1994,65(3):217-225
Gibberella fujikuroi (Fusarium moniliforme) is a complex group of plant pathogens. Some strains produce gibberellic acid and other gibberellins that promote growth and regulate various stages in plant development.The paper describes the research effort directed to development of genetic tools for this species. Furthermore the main features of the gibberellin biosynthetic pathway as established in Gibberella are described.Abbreviations AMO 1618
2-isopropyl-4-(trimethylammonium chloride)-5-methylphenylpiperidine-1-carboxylate
- hydroxykaurenoic acid
ent-kaur-16-en-7-ol-19-oic acid
- kaurenal
ent-kaur-16-en-19-al
- kaurene
ent-kaur-16-ene
- kaurenoic acid
ent-kaur-16-en-19-oic acid
- kaurenol
ent-kaur-16-en-19-ol
- paclobutrazol
1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-lyl)pentan-3-ol
- pefurazoate
pent-4-enyl-N-furfuryl-N-imidazol-1-ylcarbonyl-DL-homoa laninate
- tetcyclacis
5-(4-chlorophenyl)-3,4,5,9,10-pentaazatetracyclo-5,4,102.6,O8.11-dodeca-3,9-diene
- triarimol
-(2,4-dichlorophenyl)--phenyl-5-pyrimidine methyl alcohol 相似文献
229.
《FEBS letters》1994,350(2-3):155-158
While many ion channels are modulated by phosphorylation, there is growing evidence that they can also be regulated by Ca2+-calmodulin, apparently through direct binding. In some cases, this binding activates channels; in others, it modulates channel activities. These phenomena have been documented in Paramecium, in Drosophila, in vertebrate photoreceptors and olfactory receptors, as well as in ryanodine receptor Ca2+-release channels. Furthermore, studies on calmodulin mutants in Paramecium have shown a clear bipartite distribution of two groups of mutations in the calmodulin gene that lead to opposite behavioral and electrophysiological phenotypes. These results indicate that the N-lobe of calmodulin specifically interacts with one class of ion-channel proteins and the C-lobe with another. 相似文献
230.