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11.
Walsh H Govind AP Mastro R Hoda JC Bertrand D Vallejo Y Green WN 《The Journal of biological chemistry》2008,283(10):6022-6032
Recent evidence suggests that in addition to alpha4beta2 and alpha3-containing nicotinic receptors, alpha6-containing receptors are present in midbrain dopaminergic neurons and involved in the nicotine reward pathway. Using heterologous expression, we found that alpha6beta2, like alpha3beta2 and alpha4beta2 receptors, formed high affinity epibatidine binding complexes that are pentameric, trafficked to the cell surface, and produced acetylcholine-evoked currents. Chronic nicotine exposure up-regulated alpha6beta2 receptors with differences in up-regulation time course and concentration dependence compared with alpha4beta2 receptors, the predominant high affinity nicotine binding site in brain. The alpha6beta2 receptor up-regulation required higher nicotine concentrations than for alpha4beta2 but lower than for alpha3beta2 receptors. The alpha6beta2 up-regulation occurred 10-fold faster than for alpha4beta2 and slightly faster than for alpha3beta2. Our data suggest that nicotinic receptor up-regulation is subtype-specific such that alpha6-containing receptors up-regulate in response to transient, high nicotine exposures, whereas sustained, low nicotine exposures up-regulate alpha4beta2 receptors. 相似文献
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Breast cancer preferentially metastasizes to the skeleton, a hospitable environment that attracts and allows breast cancer cells to thrive. Growth factors released as bone is degraded support tumor cell growth, and establish a cycle favoring continued bone degradation. While the osteoclasts are the direct effectors of bone degradation, we found that osteoblasts also contribute to bone loss. Osteoblasts are more than intermediaries between tumor cells and osteoclasts. We have presented evidence that osteoblasts contribute through loss of function induced by metastatic breast cancer cells. Metastatic breast cancer cells suppress osteoblast differentiation, alter morphology, and increase apoptosis. In this study we show that osteoblasts undergo an inflammatory stress response in the presence of human metastatic breast cancer cells. When conditioned medium from cancer cells was added to human osteoblasts, the osteoblasts were induced to express increased levels of IL-6, IL-8, and MCP-1; cytokines known to attract, differentiate, and activate osteoclasts. Similar findings were seen with murine osteoblasts and primary murine calvarial osteoblasts. Osteoblasts are co-opted into creating a microenvironment that exacerbates bone loss and are prevented from producing matrix proteins for mineralization. This is the first study implicating osteoblast produced IL-6, IL-8 (human; MIP-2 and KC mouse), and MCP-1 as key mediators in the osteoblast response to metastatic breast cancer cells. 相似文献
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Endocrine studies of 4 anencephalic infants were carried out. No hypothalamic or hypophyseal structures could be found in any of them macroscopically, but we cannot say that there were no functioning pituitary cells which might have been seen microscopically. A combined LH-RH and TRH test was performed in the 6th h of life, followed by an intravenous glucose tolerance test 1 h later. Our data suggest that: (1) adenohypophyseal tissue, present in anencephaly even in the absence of a hypothalamus, is able to synthesize PRL and TSH autonomously and, under specific stimulation, PRL, TSH, and ACTH can be released while FSH and LH-HCG are not, hGH secretion is doubtful; (2) the circulating hGH and TSH surges that normally occur after delivery are hypothalamus-dependent and do not occur in anencephalics; (3) the thyroid and adrenals are able to synthesize hormones when specifically stimulated, even in the absence of the hypothalamus, and (4) beta-pancreatic function is not markedly impaired in anencephaly. 相似文献
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We report the development of a microtiter plate assay for protein kinase C. Reaction components and enzyme samples (protein kinase C purified by phosphatidylserine/cholesterol affinity or DEAE-Sephacel ion-exchange chromatography) were added to wells of a 96-well microtiter plate. The assay was started by the addition of [gamma-32P]ATP with a repeating pipet. After a 3-min incubation at 30 degrees C the wells were sampled six at a time with a 12-channel pipet and spotted onto phosphocellulose filter paper rectangles which were washed with tap water and acetone and counted for radioactivity. The microtiter plate method was more rapid than but gave results similar to those of a standard assay performed in plastic test tubes individually incubated in a 30 degrees C water bath. The microtiter plate procedure gave an intraassay (within one plate) variation of less than 9% and an interassay (between plates) variation of less than 5%. It was linear with time of incubation for 20 min and with amount of enzyme. This method can be used to expedite the assaying of column chromatography fractions for protein kinase C (and other kinase) activity. 相似文献
16.
Electroantennogram measurements of atmospheric pheromone concentration after aerial and ground application of gypsy moth mating disruptants 总被引:1,自引:0,他引:1
K. W. Thorpe J. van der Pers D. S. Leonard P. Sellers V. C. Mastro R. E. Webb R. C. Reardon 《Journal of Applied Entomology》2007,131(2):146-152
Abstract: A portable electroantennogram (EAG) sensor was used to measure relative atmospheric pheromone concentration in forest plots treated with aerial and ground applications of gypsy moth, Lymantria dispar (L.) (Lep., Lymantriidae), mating-disruption formulations. Five treatments (Disrupt II flakes with sticker, Disrupt II flakes without sticker, Disrupt II flakes in a sticker slurry, microcapsules and hand-applied Luretape), all applied at 75 g active ingredient per hectare and an untreated control were evaluated. Gypsy moth male catch in pheromone-baited traps and fertilization of deployed females were suppressed in all treatments, and no females deployed in treated plots produced more than 5% fertile eggs. Relative pheromone concentrations were significantly higher in the two treatments in which flakes were aerially applied with sticker and in the microcapsule treatment. Pheromone concentration measurements in the flakes without sticker and hand-applied treatments were not significantly different from those in the control. Mating success was negatively correlated with relative pheromone concentration. The ability of the EAG to detect differences in pheromone concentration that are correlated with mating success suggests that this could be a useful method for predicting the effectiveness of mating-disruption treatments. 相似文献
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Abstract. The high affinity form of interleukin-2 receptor (IL-2R) is composed of two subunits; the α (p55) and β (p75). The α chain, unlike the β, is expressed only on activated T lymphocytes. Therefore, high affinity binding of interleukin-2 (IL-2) is controlled by the expression of the IL-2R α-chain. In this study, we examined the effect of cytochalasin B (CB) plus 12-O-tetradecanoylphorbol-13-acetate (TPA) on expression of IL-2 and IL-2R. Northern blot and flow cytometric analysis showed that the IL-2R α-chain was expressed both at mRNA and protein levels. However, IL-2 gene expression was not induced by this treatment. Unlike the cells treated individually with CB or TPA, cells treated with CB plus TPA accumulated IL-2R mRNA at all the times examined. In order to determine the percentage of cells that incorporated tritiated thymidine ([3 H]dT) in the presence of IL-2 after treatment with CB plus TPA, autoradiography was carried out. We found that about 11% of the cells were labelled. Because the percentage of labelled cells and cells expressing IL-2R α-chain was relatively low (11% and 9% respectively), perhaps CB plus TPA caused IL-2R expression in only a subset of T cells. 相似文献
20.
Prolactin (PRL) is mitogenic for lymphocytes in vitro , but the responsiveness of lymphocytes depends on the in vivo hormonal status of the rats from which the cells were obtained. Lymphocytes from ovariectomized (OVX) rats, but not from rats in oestrus or from male rats, respond to prolactin; administration of oestradiol to OVX rats diminishes the response. In order to determine if a correlation exists between lymphocyte responsiveness to prolactin and levels of cell surface prolactin receptors (PRL-R) expression, the percentage of splenocytes and each splenocyte subpopulation expressing surface PRL-R from rats of various hormonal states (OVX, oestradiol-injected OVX, oestrus and male) was analysed by single-colour and dual-colour flow cytometric analysis. We found that approximately 20% of splenocytes expressed surface PRL-R regardless of hormonal states ( n =16). The majority (85%) of PRL-R positive splenocytes were B lymphocytes whereas 11.1% and 4.8% of splenocytes expressing the PRL-R were CD4 positive T-helper (TH ) and CD8 positive T-cytotoxic (TC ) lymphocytes, respectively. B lymphocytes also stained more brightly than T lymphocytes. This distribution of PRL-R expression did not show significant alterations on total splenocytes or TH and TC lymphocytes during various hormonal stages. However, the percentage of PRL-R-positive B lymphocytes increased markedly in OVX rats (twofold), compared to rats at oestrus. In summary, no correlation was found between the responsiveness to prolactin as a mitogen and levels of PRL-R expression by lymphocytes from rats at different hormonal states. This result suggests that sex steroid hormones may control prolactin responsiveness of lymphocytes by affecting the signal transduction pathway through PRL-R rather than by altering the level of the cell surface receptor expression. 相似文献