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A Ca2+-stimulated adenosine triphosphatase in Golgi-enriched membranes of lactating murine mammary tissue. 总被引:1,自引:0,他引:1 下载免费PDF全文
C D Watters 《The Biochemical journal》1984,224(1):39-45
A membrane fraction isolated from lactating murine mammary tissue and enriched for the Golgi membrane marker enzyme galactosyltransferase exhibited Ca2+-stimulated ATPase activity (Ca-ATPase) in 20 microM-free Mg2+ and 10 microM-MgATP, with an apparent Km for Ca2+ of 0.8 microM. Exogenous calmodulin did not enhance Ca2+ stimulation, nor could Ca-ATPase activities be detected in millimolar total Mg2+ and ATP. When assayed with micromolar Mg2+ and MgATP the Ca-ATPases of skeletal-muscle sarcoplasmic reticulum and of calmodulin-enriched red blood cell plasma membranes were half-maximally activated by 0.1 microM- and 0.6 microM-Ca2+ respectively. All three Ca-ATPases were inhibited by similar micromolar concentrations of trifluoperazine, but the Golgi activity was unaffected by quercetin in concentrations which completely inhibited both the sarcoplasmic-reticulum and red-blood-cell enzymes. The results are consistent with the hypothesis that the high-affinity Ca-ATPase is responsible for the ATP-dependent Ca2+ transport exhibited by Golgi-enriched vesicles derived from lactating mammary gland [Neville, Selker, Semple & Watters (1981) J. Membr. Biol. 61, 97-105; West (1981) Biochim. Biophys. Acta 673, 374-386]. 相似文献
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Manipulation of platelet aggregation by prostaglandins and their fatty acid precursors: pharmacological basis for a therapeutic approach 总被引:2,自引:0,他引:2
Addition of the one-, two- or three- series endoperoxide to human platelet-rich plasma tend to suppress aggregation, through the action of their respective non-enzymatic breakdown products PGE1, PGD2, or PGD3 all of which elevate cyclic AMP levels. On the other hand, these stable primary products do not arise in appreciable amounts from intrinsic endoperoxides generated from either endogenous or exogenous free fatty acids. 5,8,11,14,17-Eicosapentaenoic acid (EPA) suppresses arachidonic acid (5,8,11,14-eicosatetraenoic acid) conversion by cyclooxygenase (as well as lipoxygenase) to aggregatory metabolites in platelets. Exogenously added EPA was capable of inhibiting PRP aggregation induced either by exogenous or endogenous (released by ADP or collagen) arachidonate. The hypothetical combination of an EPA-rich diet and a thromboxane synthetase inhibitor might abolish production of the pro-aggregatory species, thromboxane A2, and enhance formation of the anti-aggregatory metabolite, prostacyclin. Whereas EPA is not detectably metabolized by platelets, dihomo-gamma-linolenic acid (8,11,14-eicosatrienoic acid) is primarily converted by cyclooxygenase and thromboxane synthetase into the inactive metabolite, 12-hydroxyheptadecadienoic (HHD) acid. Pretreatment of human platelet suspensions with the thromboxane synthetase inhibitor imidazole unmasks the aggregatory property of PGH1 and DLL which was partially compromised by the PGE1 formed. The combination of the thromboxane synthetase inhibitor and an adenylate cyclase inhibitor unmasks a complete irreversible aggregation by DLL or PGH1. The basis of a dietary strategy that replaces AA with DLL must rely on the production by the platelet of an inactive metabolite (HHD) rather than thromboxane A2. 相似文献
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Miranda N. Vesy Jessa L. Watters Raymond W. Moody Eric M. Schauber Jennie M. Mook Cameron D. Siler 《The Journal of wildlife management》2021,85(6):1267-1279
Habitat fragmentation has negative consequences on threatened and endangered species by creating isolated populations. The Texas horned lizard (Phrynosoma cornutum) is experiencing population declines and localized extirpations throughout its range and has been classified as a species of greatest conservation need in Oklahoma, USA. Younger age classes have been poorly studied but may be vital to the stability of remaining populations. To address gaps in knowledge concerning subadult (hatchling and juvenile) morphometrics, survivorship, and home range sizes, we studied 2 cohorts of subadults, for 2 years each, covering their hatching and juvenile years (2016–2019). We used a combination of radio-telemetry and novel harmonic radar methodology to study a closed population of Texas horned lizards in 15 ha of native grassland at Tinker Air Force Base, Oklahoma. Population abundance for adults and juveniles was estimated as 56.5 ± 5.5 lizards and density as 7.96 lizards/ha. Our lowest estimates of survival indicated an average survival probability for the hatchling life stage of 0.285 (95% CI = 0.15–0.44), which is lower than for adults on the site. Average home range size increased from hatchling to adult life stages. Our results will have an immediate effect on the planning and assessment of ongoing headstart and management programs for Texas horned lizards. © 2021 The Authors. The Journal of Wildlife Management published by Wiley Periodicals LLC on behalf of The Wildlife Society. 相似文献
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Intermittent hypoxia (IH) during sleep is a hallmark of sleep apnea, causing significant neuronal apoptosis, and cognitive and behavioral deficits in CNS regions underlying memory processing and executive functions. IH-induced neuroinflammation is thought to contribute to cognitive deficits after IH. In the present studies, we tested the hypothesis that IH would differentially induce inflammatory factor gene expression in microglia in a CNS region-dependent manner, and that the effects of IH would differ temporally. To test this hypothesis, adult rats were exposed to intermittent hypoxia (2 min intervals of 10.5% O2) for 8 hours/day during their respective sleep cycles for 1, 3 or 14 days. Cortex, medulla and spinal cord tissues were dissected, microglia were immunomagnetically isolated and mRNA levels of the inflammatory genes iNOS, COX-2, TNFα, IL-1β and IL-6 and the innate immune receptor TLR4 were compared to levels in normoxia. Inflammatory gene expression was also assessed in tissue homogenates (containing all CNS cells). We found that microglia from different CNS regions responded to IH differently. Cortical microglia had longer lasting inflammatory gene expression whereas spinal microglial gene expression was rapid and transient. We also observed that inflammatory gene expression in microglia frequently differed from that in tissue homogenates from the same region, indicating that cells other than microglia also contribute to IH-induced neuroinflammation. Lastly, microglial TLR4 mRNA levels were strongly upregulated by IH in a region- and time-dependent manner, and the increase in TLR4 expression appeared to coincide with timing of peak inflammatory gene expression, suggesting that TLR4 may play a role in IH-induced neuroinflammation. Together, these data indicate that microglial-specific neuroinflammation may play distinct roles in the effects of intermittent hypoxia in different CNS regions. 相似文献
7.
Jonathan Griffin Darren C. Watters Hunan Yi Ahmed Iraqi David Lidzey Alastair R. Buckley 《Liver Transplantation》2013,3(7):903-908
Bulk heterojunction solar cells containing molybdenum oxide hole extracting anode contacts have been fabricated with varying stoicheometry using radio frequency reactive sputtering from a Molybdenum metal target. A blend of the newly synthesised conjugated polymer poly[9‐(heptadecan‐9‐yl)‐9H‐carbazole‐2,7‐diyl‐alt‐(5,6‐bis(octyloxy)‐4,7‐di(thiophen‐2‐yl)benzo[c][1,2,5]thiadiazole)‐5,5‐diyl] (PCDTBT‐8) and fullerene [6,6]‐Phenyl‐C71‐butyric acid methyl ester (PC70BM) was used as the photoactive layer and device results show that anodes with greater than 98% Molybdenum (VI) oxide result in peak power conversion efficiencies of 3.7%.The presence of up to 28% of Mo (V) results in no significant reduction in efficiency, however the presence of metallic Mo (IV) and lower oxidation states lead to severe reductions in device performance due to a combination of a large hole extraction energy barrier of approximately 0.9eV and reduced device stability. 相似文献
8.
Batool Shannan Quan Chen Andrea Watters Michela Perego Clemens Krepler Rakhee Thombre Ling Li Geena Rajan Scott Peterson Phyllis A. Gimotty Melissa Wilson Katherine L. Nathanson Tara C. Gangadhar Lynn M. Schuchter Ashani T. Weeraratna Meenhard Herlyn Adina Vultur 《Pigment cell & melanoma research》2016,29(3):317-328
Targeted therapies for mutant BRAF metastatic melanoma are effective but not curative due to acquisition of resistance. PI3K signaling is a common mediator of therapy resistance in melanoma; thus, the need for effective PI3K inhibitors is critical. However, testing PI3K inhibitors in adherent cultures is not always reflective of their potential in vivo. To emphasize this, we compared PI3K inhibitors of different specificity in two‐ and three‐dimensional (2D, 3D) melanoma models and show that drug response predictions gain from evaluation using 3D models. Our results in 3D demonstrate the anti‐invasive potential of PI3K inhibitors and that drugs such as PX‐866 have beneficial activity in physiological models alone and when combined with BRAF inhibition. These assays finally help highlight pathway effectors that could be involved in drug response in different environments (e.g. p4E‐BP1). Our findings show the advantages of 3D melanoma models to enhance our understanding of PI3K inhibitors. 相似文献
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Denlinger LC Sommer JA Parker K Gudipaty L Fisette PL Watters JW Proctor RA Dubyak GR Bertics PJ 《Journal of immunology (Baltimore, Md. : 1950)》2003,171(3):1304-1311
Activation of the P2X(7) receptor by extracellular nucleotides modulates multiple immune functions, including inflammatory mediator production, membrane fusion events, and apoptosis. Previous studies have revealed that the C terminus of this multimeric cation channel possesses a lipid-interaction motif that has been proposed to regulate receptor function. This domain is homologous to the LPS binding region of the LPS binding protein, and we demonstrated that two basic residues (Arg(578), Lys(579)) within this motif are essential for LPS binding to P2X(7) in vitro. Because P2X(7) can influence LPS action, and because lipid interaction motifs modulate the trafficking of other ion channel-linked receptors, we hypothesized that this motif of P2X(7) is critical for receptor function and trafficking. In these studies we mutated Arg(578) and Lys(579) of P2X(7), and the expression profile, channel activity, and pore formation of the mutant were characterized in transfected human embryonic kidney 293 cells. In contrast with the wild-type receptor, the P2X(7)-R578E/K579E mutant fails to demonstrate surface immunoreactivity despite normal levels of total protein expression. This effect on the mutant receptor is unlikely to result from widespread defects in protein folding, because surface localization, determined using conformation-specific Abs, can be restored by growing the cells at 25 degrees C, conditions that slow receptor recycling. Despite surface expression at reduced temperatures, at 25 degrees C the P2X(7)-R578E/K579E mutant still exhibits greatly reduced sodium, potassium, and calcium channel activity when compared with the wild-type receptor, and cannot induce pore formation. These data suggest that the lipid interaction motif of the P2X(7) C terminus controls receptor trafficking and modulates channel activity. 相似文献