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101.
Summary To date, the small size and slow growth of eosinophil colonies in vitro has hampered study of cloned eosinophils. We found
enhanced eosinophil colony size and numbers in methylcellulose cultures of bone marrow cells utilizing defined supplemented
bovine calf serum (DSBCS) in combination with EL4 conditioned medium (EL4-CM). At days 9, 16 and 23 significantly more eosinophil
colonies and more cells/colony were present in cultures incubated with DSBCS/EL4-CM than in cultures incubated with fetal
calf serum/EL4-CM. The ability to generate large numbers of eosinophils in vitro should facilitate study of cloned eosinophils.
Supported in part by a grant from the National Institutes of Health, AI 20416, and by the Mayo Foundation.
Editor's statement Previous approaches to in vitro culture of eosinophils generally have achieved, at best, mixed cultures
of colonies of these cells and granulocyte-macrophage colonies. The improved culture methods described in this report produce
more homogeneous eosinophil cultures and larger colonies of these cells. The procedure employs EL4 murine thymoma-conditioned
medium, which apparently contains eosinophil colony-stimulating activity in the absence of granulocyte-macrophage colony-stimulating
activity. David W. Barnes 相似文献
102.
103.
Cell suspension cultures of Corydalis sempervirens have provenideal for the study of fusicoccin action [Schulz et al. (1990)Planta 183: 83] and express the fusicoccin-binding protein aswell as a plasma membrane H+-ATPase which is activated by thefungal toxin. Microsomal vesicles prepared from these cellsaccumulate Ca2+ in the presence of Mg-ATP. The protonophorecar-bonylcyanide m-chlorophenylhydrazone did not inhibit theMg-ATP dependent Ca2+-transport into the vesicles. This processis thus due to the activity of at least one primary active,ATP-driven, Ca2+-pump. The enzyme was characterized in detail.It has a pH optimum of 7.2, an apparent Km of 0.3 mu (ATP),12pm (Ca2+), accepts ATP>ITP GTP>CTP UTP, and is strongly(Ki, app 0.75 µmM) inhibited by erythrosine B but lessso (Ki, app 95 µM) by or-thovanadate. These characteristicsare typical for the plasma membrane Ca2+-ATPase characterizedfrom differentiated tissues [Graf and Weiler (1990) Physiol.Plant. 75: 634]. Fusicoccin activates the erythrosine-sensitiveCa2+-pump by lowering its Km for ATP, when added to living cellsprior to tissue homogenization. Thus, fusicoccin appears toactivate at least two ion-translocating ATPases in one and thesame tissue, suggesting that the toxin's mechanism of actionis complex and not restricted to activation of the H+-ATPase.FC has no effect when administered to microsomes. The microsomalenzyme was solubilized and reconstituted into asolec-tin liposomesin functional form. The reconstituted, erythrosine sensitiveCa2+-ATPase was insensitive to fusicoccin. Thus, componentsessential for toxin action are either lost or inactivated duringsubcellular fractionation. It is likely that FC action requiressoluble components. (Received April 22, 1991; Accepted July 24, 1991) 相似文献
104.
Peter James O'Brien Hua Shen Janice Weiler Mehdi Mirsalimi Richard Julian 《Molecular and cellular biochemistry》1991,102(1):1-12
A novel, simple, rapid and reproducible microassay is used for kinetic analysis of Ca-sequestration by homogenates of myocardium of turkeys with furazolidone-induced congestive cardiomyopathy. The assay monitors Ca in real-time using dual-emission ratiometric spectrofluorometry and the Ca-indicator dye indo-1. Using this assay and isolated SR studies we make several novel findings regarding the mechanism of SR failure in furazolidone cardiomyopathy.Qualitative differences in Ca-sequestration were not detected between groups. However, compared to controls the furazolidone treatment resulted in: 1) 50% depression in maximal activities (1.54 ± 0.36 vs 0.73 ± 0.12 µM/sec); 2) 2-fold increases in post-sequestration concentrations of ionized Ca(79 ± 23 vs 141 ±13 nmol Ca/L homogenate); 3) 2-fold increases in Ca half-life (415 vs 790 msec); and 4) 25% increased passive Ca-binding capacity of homogenates. The Ca-ATPase specific activity of isolated sarcoplasmic reticulum was 60% increased in congestive cardiomyopathy (543 ± 140 vs 873 ± 108 nmol ATP hydrolyzed/min/mg membrane protein) although membrane yield was 20% decreased (0.79 ± 0.09 vs 0.63 ± 0.03 mg/g heart). The increased ATPase and decreased Ca-uptake activities in combination with the occurrence of 36% cardiac hypertrophy and 19% decreased body weights resulted in estimates of the relative energy cost to the animal for myocardial Ca transport being 5.5-fold increased with cardiomyopathy (20.5 vs 111 nmol ATP hydrolyzed per µM decrease of sarcoplasmic free Ca/kg body weight).These data indicate that congestive cardiomyopathy is associated with markedly increased permeability of sarcoplasmic reticulum to Ca and compensatorily increased Ca-ATPase activity. Accelerated energy consumption due to the increased energy cost of Ca transport and increased time of myocyte activation are predicted to predispose the myocardium to fatigue and irreversible failure. 相似文献
105.
106.
107.
Weiler PG 《The Western journal of medicine》1988,148(2):199-200
108.
K. W. Schmid R. Weiler R. W. Xu R. Hogue-Angeletti R. Fischer-Colbrie H. Winkler 《The Histochemical journal》1989,21(6):365-372
Summary Antisera were raised against synthetic peptides derived from the primary amino acid sequence of human chromogranin B. These antisera recognized in one- and two-dimensional immunoblotting a component previously designated as chromogranin B. In human chromaffin granules, the major endogenous processing product of chromogranin B is formed by proteolytic cleavage of the protein near theC-terminus. Immunohistochemical localizations were obtained with antisera against human chromogranins A and B and against a synthetic peptide corresponding to the B sequence. In human tissues, chromogranin B is co-stored with chromogranin A in the adrenal medulla, the anterior pituitary, parafollicular cells of the thyroid, in some cells of the endocrine pancreas and in some enterochromaffin cells, whereas only chromogranin A is found in the parathyroid gland and enterochromaffin cells of the gastric corpus mucosa. In the nervous system, no immunostaining was observed for chromogranin A and only a weak one for chromogranin B in some cells of the spinal cord. However, the Purkinje cells of the cerebellum were strongly positive for chromogranin B. 相似文献
109.
E. W. Weiler 《Plant biology (Stuttgart, Germany)》1993,106(1):2-4
Recent evidence suggests that octadecanoid derived metabolites, notably jasmonic acid and/or methyl jasmonate, are involved in the reaction of touch-sensitive tendrils to mechanical stimulation. 相似文献
110.
R. R. Weiler 《Hydrobiologia》1988,163(1):77-83
Intensive surveys of Georgian Bay and the North Channel were done in 1974 and 1980. This paper reviews the main findings of the surveys and describes the main factors that determine the chemistry of the two water bodies. Georgian Bay is oligotrophic. Its chemistry is determined chiefly by natural phenomena, specifically the geology of the drainage basin and the river runoff in the spring, and by exchange with Lake Huron and the North Channel. Localized eutrophication is found in the bays along the southern shore. The chemistry of the North Channel is determined by its very short residence time — about two years — caused by the large inflow from St. Mary's River. The water quality in the river reflects that of Lake Superior and the inputs of nutrients and contaminants from Sault Ste. Maire. Because of the short flushing time, the Channel remains oligotrophic despite the large loadings of phosphorus. 相似文献