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OBJECTIVE: The aim was to examine whether mitochondrial Ca2+ fluxes are high enough to change mitochondrial and cytosolic calcium concentration during the contraction cycle. METHODS: Isolated guinea pig ventricular myocytes were stimulated with paired voltage clamp pulses until contractions were maximal (2 mM [Ca2+]o, 36 degrees C). At defined times of diastole or systole, the cells were shock frozen. Electron-probe microanalysis measured the concentration of total calcium in mitochondria (sigma Ca(mito)) and surrounding cytosol (sigma Cac). Other experiments were performed to evaluate DNP sensitive mitochondrial Ca2+ uptake from depolarisation induced [Ca2+]c transients (K5indo-1 fluorescence). RESULTS: At end of diastole, sigma Ca(mito) was 446 mumol.litre-1. During systole, sigma Ca(mito) increased with a 20 ms delay. A peak sigma Ca(mito) of 1050 mumol.litre-1 was measured 40 ms after start of systole, while 95 ms after start of systole sigma Ca(mito) had fallen to 530 mumol.litre-1. From the changes in sigma Ca(mito) the rates of net mitochondrial Ca2+ flux were estimated at 100 nmol.s-1 x mg-1 protein for Ca2+ influx and 36 nmol.s-1 x mg-1 protein for Ca2+ egress. Decay of sigma Ca(mito) was coupled to a rise in sigma Na(mito). sigma Cl(mito) and sigma K(mito) rose and fell in parallel with sigma Ca(mito), suggesting Ca2+ activation of mitochondrial anion and cation channels. Activation of the non-specific permeability can be excluded. Block of mitochondrial Ca2+ uptake with DNP (100 microM) or FCCP (10 microM) increased the amplitude of the [Ca2+]c transients for 1-3 min by about 50%; evaluation of mitochondrial Ca2+ uptake from DNP sensitive difference signals, however, was hampered by sequestration of mitochondrial Ca2+ into the sarcoplasmic reticulum. CONCLUSIONS: Mitochondrial calcium content changes during each individual contraction cycle; a substantial amount of calcium is taken up during the systole and released during later systole and diastole.  相似文献   
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Enterprises increasingly recognize the compelling economic and operational benefits from virtualizing and pooling IT resources in the cloud. Nevertheless, the significant and valuable transformation of organizations that adopt cloud computing is accompanied by a number of security threats that should be considered. In this paper, we outline significant security challenges presented when migrating to a cloud environment and propose PaaSword – a novel holistic framework that aspires to alleviate these challenges. Specifically, the proposed framework involves a context-aware security model, the necessary policies enforcement mechanism along with a physical distribution, encryption and query middleware.  相似文献   
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An attempt was made to clarify whether the site of postchiasmal lesions affects subjective perception of homonymous visual field defects during stimulation with flickering random dot patterns (white noise-field). Out of 56 patients with homonymous hemianopia, 38 (68%) perceived scotomata in this situation, but 18 (32%) discerned none at all. Neuroradiologic superposition of cerebral lesions detected by computed tomography (CT) or magnetic resonance imaging (MRI) showed that nearly all patients who perceived their scotomata had lesions involving the primary visual cortex or the perigeniculate region, whereas those who received no scotoma had lesions centered within the optic radiation. Functional MRI of six normal subjects during stimulation with flickering random dot patterns indicated predominant activation of the primary visual cortex. Since noise-field defects were most frequently perceived by patients whose lesion involved the primary visual cortex, it appears that the sensitivity of noise-field campimetry depends on the site of damage in the visual pathway. The explanation for this may be that damage to long-range horizontal connections impairs filling-in processes.  相似文献   
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The analysis of airborne hyperspectral data is often affected by brightness gradients that are caused by directional surface reflectance. For line scanners these gradients occur in across-track direction and depend on the sensor's view-angle. They are greatest whenever the flight path is perpendicular to the sun-target-observer plane. A common way to correct these gradients is to normalize the reflectance factors to nadir view. This is especially complicated for data from spatially and spectrally heterogeneous urban areas and requires surface type specific models. This paper presents a class-wise empirical approach that is adapted to meet the needs of such images.Within this class-wise approach, empirical models are fit to the brightness gradients of spectrally pure pixels from classes after a spectral angle mapping (SAM). Compensation factors resulting from these models are then assigned to all pixels of the image, both in a discrete manner according the SAM and in a weighted manner based on information from the SAM rule images. The latter scheme is designed in consideration of the great number of mixed pixels.The method is tested on data from the Hyperspectral Mapper (HyMap) that was acquired over Berlin, Germany. It proves superior to a common global approach based on a thorough assessment using a second HyMap image as reference. The weighted assignment of compensation factors is adequate for the correction of areas that are characterized by mixed pixels.A remainder of the original brightness gradient cannot be found in the corrected image, which can then be used for any subsequent qualitative and quantitative analyses. Thus, the proposed method enables the comparison and composition of airborne data sets with similar recording conditions and does not require additional field or laboratory measurements.  相似文献   
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Fifty-one thyroid tumours and tumour-like lesions were analysed for instability at ten dinucleotide microsatellite loci and at two coding mononucleotide repeats within the transforming growth factor beta (TGF-beta) type II receptor (TbetaRII) and insulin-like growth factor II (IGF-II) receptor (IGFIIR) genes respectively. Microsatellite instability (MI) was detected in 11 out of 51 cases (21.5%), including six (11.7%) with MI at one or two loci and five (9.8%) with MI at three or more loci (RER+ phenotype). No mutations in the TbetaRII and IGFIIR repeats were observed. The overall frequency of MI did not significantly vary in relation to age, gender, benign versus malignant status and tumour size. However, widespread MI was significantly more frequent in follicular adenomas and carcinomas than in papillary and Hürthle cell tumours: three out of nine tumours of follicular type (33.3%) resulted in replication error positive (RER+), versus 1 out of 29 papillary carcinomas (3.4%, P = 0.01), and zero out of eight Hürthle cell neoplasms. Regional lymph node metastases were present in five MI-negative primary cancers and resulted in MI-positive in two cases.  相似文献   
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