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1.
1. The tRNA preparation obtained from human placenta by phenol extraction and DEAE-cellulose chromatography was homogeneous on ultracentrifugation and showed high amino acid acceptor activity. 2. The analysis of the isoaccepting tRNA by reversed-phase chromatography (RPC-5 system) showed the presence of 5 fractions for glycine and leucine, 3 for tyrosine, alanine and valine, 2 for arginine and 1 for phenylalanine.  相似文献   

2.
An equation of state and expressions for the isothermal compressibility, thermal expansion coefficient, heat capacity and entropy of liquids have been derived from the Free Volume Model partition function suggested by Turnbull. The simple definition of the free volume is used and it is assumed that the specific volume is directly related to the cube of the inter-molecular separation by a proportionality factor which is found to be a function of temperature and pressure as well as specific volume. When values of the proportionality factor are calculated from experimental data for real liquids, it is found to be approximately constant over ranges of temperature and pressure which correspond to the dense liquid phase. This result provides a single-parameter method for calculating dense liquid thermodynamic properties and is consistent with the fact that the Free Volume Model is designed to describe liquids near the solidification point. Calculated values ofβ, α, C p andS for nine liquids are compared to experimental data when possible, and agree with these data within an order of magnitude.  相似文献   

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We have discovered two substituted 4-aminopiperidine compounds having high in vitro affinity and selectivity for the human dopamine D1 receptor. Both compounds, 3-ethoxy-N-methyl-N-[1-(phenylmethyl)-4-piperidinyl]-2-pyridinylamine (U-99363E), and its 3-isopropoxy analog (U-101958), were found through a routine receptor binding screen. The determined affinities (Ki) of these compounds for the cloned human dopamine D4 receptor were 2.2 and 1.4 nM, respectively. They exhibited at least 100-fold lower affinities for dopamine D2 and for other dopaminergic, serotonergic and adrenergic receptors. Both compounds were found to antagonize quinpirole-induced mitogenesis in Chinese hamster ovary cells expressing the human dopamine D4 receptor. In spite of their poor metabolic stability and low bioavailability. U-99363E and U-101958 appear to be among the first high-affinity, highly selective dopamine D4 receptor antagonists reported, and may have utility in in vitro investigations requiring selective tagging or blockade of dopamine D4 sites.  相似文献   

5.
The present paper reviews current knowledge of the development and plasticity of the inhibitory gamma-aminobutyric acid (GABA) containing circuitry of the cerebral neocortex, in particular, the rat somatosensory barrel field cortex. Recent studies reveal a delayed and protracted maturation of the inhibitory compared with the excitatory cortical system, both at the neuronal and synaptic levels. This characteristic developmental pattern leaves a longer time window during which behaviourally relevant activity coming from the periphery can influence the organization of the GABA system. Indeed, sensory deprivation experiments confirm the involvement of the GABA system in phenomena of experience-dependent cortical plasticity. Changing the pattern and level of afferent activity of in the rat somatosensory system during development by removing vibrissae results in a significant decrease in the number of GABA neurons and synapses in the thalamocortical recipient layer IV. Particularly affected are GABA synapses contacting dendritic spines, the number of which decreases by almost two-thirds. The involvement of the GABA system in events of experience-dependent plasticity contributes to the adequate functioning of the cerebral cortex in the conditions of constantly changing environment and varying individual experience.  相似文献   

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Human placental mitochondria prepared by a new isolation procedure exhibit low but well coupled rates of state 3 respiration with different substrates (succinate: 32.3 nmol O2/mg/min, RCI = 4.4; pyruvate: 12.6 nmol O2/mg/min, RCI R = 4.2; palmitoylcarnitine: 16.6 nmol O2/mg/min, RCI R = 4.9). The addition of the uncoupler FCCP increased the respiratory rates (succinate: 40.7 nmol O2/mg/min; pyruvate: 21.2 nmol O2/mg/min: palmitoylcarnitine: 25.4 nmol O2/mg/min). The low respiratory rates correlate well with a low capacity of the respiratory chain as shown by the specific contents of cytochrome c (0.15 nmol/mg), cytochrome b (0.19 nmol/mg) and cytochrome oxidase (0.14 nmol/mg) as well as with the low content of adenine nucleotides (2.71 nmol/mg). These data together with the finding of high activities of alkaline phosphatase (2.2 U/mg) support the view that human placental mitochondria are contaminated with nonmitochondrial membranes. Since it was not possible to obtain functionally intact mitochondria with negligible activities of alkaline phosphatase the influence of this enzyme on the extramitochondrial adenine nucleotide turnover was investigated. Alkaline phosphatase splits phosphate from ATP, ADP and AMP with different rates resulting in an intermediate accumulation of AMP. Mitochondrial adenylate kinase (0.16 U/mg) regenerated ADP from AMP and ATP resulting in drastically decreased ADP/O ratios and prolonged state 3 respirations. Inhibiting the adenylate kinase with diadenosine pentaphosphate the ADP regeneration from AMP and ATP was suppressed which, in turn, enhanced the ADP/O ratios. In the absence of magnesium ions, if both the alkaline phosphatase and the adenylate kinase are inhibited normal ADP/O ratios and state 3-state 4 transitions can be observed. Under these conditions, human placental mitochondria showed normal properties comparable to those of mitochondria from other tissues with the only exception of low specific activities.  相似文献   

8.
探究了菌渣的水热液化转换成生物油燃料的过程。结果表明,抗生素菌渣在260 ℃、保留时间是135 min时,获得最大的生物油产率(28.01%)。通过6种不同的催化剂进行催化,加入催化剂后,生物油产率最大的是Na2CO3(36.06%)和NaOH(36.31%)。碱催化的生物油的含氮化合物的质量分数在41.16%~49.74%之间,而酸催化产生的生物油含氮化合物的量在57.62%~59.32%之间。通过调节催化剂Na2CO3、NaOH的添加量发现,在投加量为8%时,生物油含氮量均最低,Na2CO3和NaOH催化产生的生物油组分的含氮化合物质量分数分别为29.12%和35.67%。在催化剂投加量为10%时,对氧的脱除效果都最好,分别为32.12%和29.02%,此时产生的生物油的热值达到最大(达到33.3220和34.7320 MJ?kg?1)。   相似文献   

9.
Cytochrome P-450 from rabbit pulmonary microsomes was purified approximately 32-fold. The purification method involved solubilization of microsomes using sodium cholate, and recovery of cytochrome P-450 in the precipitate formed between 25 to 42% saturation of the digested microsomes with ammonium sulfate in the absence of glycerol. Further purification was achieved by chromatography on DEAE-cellulose and hydroxylapatite using Emulgen 913 as an eluent. Partially purified preparations containing up to 7.4 nmol of cytochrome P-450 per mg of protein were essentially free of NADPH-cytochrome c reductase activity and cytochromes b5 and P-420. However, epoxide hydrase was found to co-purify with cytochrome P-450. The CO-difference spectrum of dithionite-reduced purified cytochrome showed the expected peak at 450 nm. However, the magnitude of the peak was dependent on added microsomal lipid fraction in the assay medium. Purified pulmonary cytochrome P-450 formed typical types I and II substrate difference spectra with benzphetamine and pyridine, respectively. Sodium dodecyl sulfate-gel electrophoresis of partially purified cytochrome P-450 gave two major bands when stained with Coomassie blue. The faster moving band which contained peroxidase activity had an estimated molecular weight of 49,000 +/- 1,200. The cytochrome P-450 fraction, when combined with solubilized pulmonary microsomal NADPH-cytochrome c reductase and lipid fractions, was active in the O-deethylation of 7-ethoxycoumarin and the N-demethylation of benzphetamine.  相似文献   

10.
Hydroxy apatite (HA) is effectively used as a bioimplant material because it closely resembles bone apatite and exhibits good biocompatibility. So, in this research, HA powders were produced by calcinations of natural bones including human, bovine, camel and horse bones, and also via sol-gel method. Powders characterizations of natural HA and Synthetic HA were studied by X-ray powder diffraction (XRD), X-ray fluorescence (XRF), Fourier transform infrared (FTIR) and scanning electron microscopy (SEM), combined with transmission electron microscopy (ТЕМ). In order to verify the biocompatibility of these HA powders, MTT assay was applied. The XRD results showed that the HA powders were successfully produced by using different sources. Also, it was obvious from XRF analysis that the main components of them were Ca and P. Furthermore, it was seen that the size of particles was in the nanometric scale and they showed agglomerates consisting of numerous nanocrystals. FTIR spectra of all samples proved the presence of various CO 3 2- , PO 4 3- and OH groups in the powders. In addition, the MTT assay revealed that the cells proliferations in the presence of horse and human HA nanopowders were stimulated.  相似文献   

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Isolation of highly purified and highly molecular monoamine oxidase (MAO) from pig liver mitochondria have been worked out. Specific activity of isolated preparation is 2700 times higher than of original mitochondria homogenate. Enzyme solubilization by digitonin, affinity chromatography purification and ultrafiltration underlie this method. MAO catalytic properties changing during the process of purification by different methods have been investigated. Substrate specificity was studied; kinetic parameters of enzymatic desemination were calculated.  相似文献   

14.
The biochemical and immunocytochemical properties of peroxisomal and mitochondrial beta-oxidation enzymes in bovine adrenal chromaffin cells were investigated. Peroxisomes were detectable by immunofluorescence staining using antibodies against acyl-CoA oxidase, peroxisomal 3-ketoacyl-CoA thiolase and catalase. The mitochondria were abundantly stained with antibody against mitochondrial 3-ketoacyl-CoA thiolase. The biosynthesis and intracellular processing of acyl-CoA oxidase and the peroxisomal 3-ketoacyl-CoA thiolase was slower than that in fibroblasts. The peroxisomal beta-oxidation activities shown by [1-14C] lignoceric acid oxidation were slightly lower than those in fibroblasts, whereas the mitochondrial beta-oxidation activities shown by [1-14C] palmitic acid oxidation were almost identical to those in fibroblasts. Adrenal chromaffin cells are useful materials for investigating the peroxisomal and mitochondrial metabolism of autonomic neurons and may contribute to the clarification of neuronal dysfunction in peroxisomal and mitochondrial disorders.  相似文献   

15.
The purpose of the present investigation was to evaluate 99Tcm-labelled alpha-D-glucose 1-phosphate (GP) aerosols for single photon emission computed tomographic (SPECT) ventilation lung imaging in comparison to 99Tcm-diethylenetriaminepentaacetate (DTPA) aerosols. Ten normal nonsmoking male volunteers (aged 20-30 years) were included in this study after obtaining their informed consent. 99Tcm-GP, 30 mCi, in 2 ml was placed in the nebulizer (Venticis II) and inhalation continued for 5 min of normal breathing with oxygen flowing through. In 10 subjects dynamic images were obtained from the posterior position for 90 min with 45 frames on a 64 x 64 matrix by the use of a gamma camera. At the end of the dynamic study planar images of the lung (anterior, posterior and laterals) were recorded. Decay corrected clearance curves and kep values were obtained by the pulmonary epithelial programme and T1/2 values were calculated. The same procedure was followed by the use of 99Tcm-DTPA in the same subjects 2 weeks later. SPECT studies of the lung were performed in five subjects after inhalation of 99Tcm-GP aerosols. Clearance curves were monoexponential. The difference in T1/2 values between the right and left lungs was statistically insignificant (P > 0.10). The mean T1/2 values were 316.5 +/- 44.7 and 80.8 +/- 13.4 min for 99Tcm-GP and 99Tcm-DTPA, respectively. The difference was significant (P < 0.0005). On scintigraphic images 99Tcm-GP showed high alveolar deposition and low adhesion to major airways like 99Tcm-DTPA. However, it is preferred to 99Tcm-DTPA for SPECT studies because of its prolonged pulmonary clearance.  相似文献   

16.
Factor I deficiency causes a permanent, uncontrolled activation of the alternative pathway resulting in an increased turnover of C3 and consumption of factor B, factor H and properdin. Factor I deficiency is clinically associated with recurrent bacterial infections already in early infancy, mainly affecting the upper and lower respiratory tract, or presenting as meningitis or septicemia. We here report on a Brazilian family (n = 9) with known consanguinity, where in 3/7 children, suffering from chronic otitis, meningitis, and respiratory infections, a complete factor I deficiency was recognized. One of the patients died after fulminant sepsis. Hemolytic activity of the alternative pathway was not detectable in the patients' sera due to decreased plasma concentrations of C3, factor B and properdin. As a consequence of factor I deficiency, C3b could not be metabolized with the result that no C3-derived split products (C3dg/C3d) were detectable in the patients' sera. In vitro reconstitution with purified factor I restored the regulatory function in the patients' sera with the subsequent cleavage of C3b to C3c and C3dg. Factor H levels were decreased in all patients' sera and found to be tightly complexed with C3b resulting in a modified electrophoretic mobility. Upon factor I reconstitution, factor H was released from C3b regaining its beta 1 electrophoretic mobility. Complement-mediated biological functions like opsonization of bacteria, chemotactic activity and phagocytosis in these patients were impaired. The parents (cousins, 2nd degree) and 3/4 siblings had significantly reduced factor I plasma levels without further alteration in their complement profile. 3 of these obviously heterozygously deficient family members suffered from recurrent bacterial infections of different frequency and severity.  相似文献   

17.
X-ray (CuKalpha) scattering curves of rat liver mitochondria are characterized by continuously decreasing intensity from 0.5 to 5 mrad and a broad maximum centered near 20 mrad. The condensed-to-orthodox morphological transition of the inner membranes of intact mitochondria causes a dramatic decrease in scattering at very small angle and a marked shift of the 20 mrad maximum to smaller angle. A similar small-angle scattering maximum is observed with inner mitochondrial membrane fractions prepared by digitonin treatment and osmotic shock/step gradient centrifugation procedures. However, the small-angle X-ray scattering curves of mitochondria after acetone treatment and osmoticlysis/sonication are essentially continuous. These characteristics of mitochondrial X-ray scattering are discussed in terms of known structural features of the organelle.  相似文献   

18.
This paper discusses the general efficacy of the rat as a model for studying the pathogenicity of respirable fibres in humans. Rats show differences in lung structure from man and these could influence the pulmonary response to some degree, but they do develop interstitial fibrosis, carcinoma and mesothelioma when exposed to respirable fibres and these same types of lesions are found in fibre-exposed humans. Because of the importance of long fibres in causing pathological change, a substantial number of long respirable fibres must be used in inhalation studies if the relative pathogenicity of different fibre types is to be assessed. From data on the fibre burden in human disease and information on build-up of fibres in rat studies in a range of different fibres, a 1 year exposure to a cloud of 200 fibres ml-1 minimum, preferably more, is recommended. The rate of dissolution of the fibres within the lung milieu will be an an important factor in determining the pathogenicity. Many questions remain unanswered as to the mechanism of lung pathogenicity caused by respirable fibres particularly in the areas of durability, fibre shape, fibre surface chemistry and the exact mechanism, at the cellular level whereby fibres can cause pathological change. The use of rat inhalation studies, properly conceived and designed, remain a key approach whereby these questions can be answered.  相似文献   

19.
The perceptual strategies used by 4 orangutans (2 subadults, 2 adults) when choosing the larger of 2 volumes in a Piagetian conservation task were investigated. Three possible perceptual strategies were investigated: (a) direct perceptual estimation of the container's content independent of its shape, (b) use of the spatial and temporal cues provided by the pouring of liquid from one container to another, and (c) ability to initially identify the larger volume and track it across transformations disregarding misleading perceptual cues. Results indicated that the direct perceptual estimation strategy was the best candidate to explain the orangutan's systematic choice of the larger of 2 quantities.  相似文献   

20.
Ethanol intake depletes the mitochondrial pool of reduced glutathione (GSH) by impairing the transport of GSH from cytosol into mitochondria. S-Adenosyl-L-methionine (SAM) supplementation of ethanol-fed rats restores the mitochondrial pool of GSH. The purpose of the current study was to determine the effect of ethanol feeding on the kinetic parameters of mitochondrial GSH transport, the fluidity of mitochondria, and the effect of SAM on these changes. Male Sprague-Dawley rats were fed ethanol-liquid diet for 4 weeks supplemented with either SAM or N-acetylcysteine (NAC). SAM-supplementation of ethanol-fed rats restored the mitochondrial GSH pool but NAC administration did not. Kinetic studies of GSH transport in isolated mitochondria revealed two saturable, adenosine triphosphate (ATP)-stimulated components that were affected significantly by chronic ethanol feeding: lowering Vmax (0.22 and 1.6 in ethanol case vs. 0.44 and 2.7 nmol/15 sec/mg protein in controls) for both low and high affinity components with the latter showing an increased Km (15.5 vs. 8.9, mmol/L in ethanol vs. control). Mitochondria from SAM-supplemented ethanol-fed rats showed kinetic features of GSH transport similar to control mitochondria. Determination of membrane fluidity revealed an increased order parameter in ethanol compared with control mitochondria, which was restricted to the polar head groups of the bilayer and was prevented by SAM but not NAC supplementation of ethanol-fed rats. The changes elicited in mitochondria by ethanol were confined to the inner membrane; mitoplasts from ethanol-fed rats showed features similar to those of intact mitochondria such as impaired transport of GSH and increased order parameter. A different mitochondrial transporter, adenosine diphosphate (ADP)/ATP translocator, was unaffected by ethanol feeding. Furthermore, fluidization of mitochondria or mitoplasts from ethanol-fed rats by treatment with a fatty acid derivative restored their ability to transport GSH to control levels. Thus, ethanol-induced impaired transport of GSH into mitochondria is selective, mediated by decreased fluidity of the mitochondrial inner membrane, and prevented by SAM treatment.  相似文献   

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