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1.
Abstract: The relative cerebral cortical metabolism of glucose (GLU) and 2-deoxy-D-glucose (DG) was measured in vivo in control and insulin-treated hypoglycemic rats. The ratio of the utilization rate constants for the two hexoses, i.e., K DG/ K CLU is defined as the Hexose Utilization Index (HUI). The HUI was found to be invariant in rats whose cerebral glucose content exceeded 1 μmo1.g−1 wet weight (HUI = 0.48 ± 0.07). Severe hypoglycemia (plasma glucose <2 mM) effected a shift in the HUI to 1.04 ± 0.21. The results are consistent with a model in which the interpretation of the HUI is determined by the rate of transport into brain, or subsequent phosphorylation, as the rate-limiting step for hexose utilization.  相似文献   

2.
High- and Low-Affinity Transport of D-Glucose from Blood to Brain   总被引:21,自引:19,他引:2  
Abstract: Measurements of the unidirectional blood-brain glucose flux in rat were incompatible with a single set of kinetic constants for transendothelial transport. At least two transfer mechanisms were present: a high-affinity, low-capacity system, and a low-affinity, high-capacity system. The low-affinity system did not represent passive diffusion because it distinguished between D-and L-glucose. The Tmax and K m, for the high-affinity system were 0.16 mmol 100 g−1 min−1 and 1 mM; for the low-affinity system, ∼ 5 mmol 100 g−1 min−1 and ∼ 1 M. With these values, physiological glucose concentrations were not sufficient to saturate the low-affinity system. In normoglycemia, therefore, three independent pathways of glucose transport from blood to brain appear to exist: a high-affinity facilitated diffusion pathway of apparent permeability 235·10−7 cm s−1, a specific but nonsaturable diffusion pathway of permeability 85·10−7 cm s−l, and a nonspecifc passive diffusion pathway of permeability 2·10−7 cm s−1.  相似文献   

3.
The values of Michaelis–Menten constant (KM) and maximum velocity (VMAX) for kidney and heart monoamine oxidase (MAO) from pacu Piaractus mesopotamicus were determined. The mean ± s . e . KM values were 17·28 ± 2·27 μM for kidney and 15·38 ± 1·86 μM for heart. MAO activities were 111·60 ± 3·25 and 15·12 ± 0·30 nmols min−1 g−1 of wet tissue for kidney and heart, respectively. In addition, MAO inhibitory studies in these two tissues indicate that this enzyme may be a different isoform of MAO.  相似文献   

4.
Abstract: Recent studies indicate the lumped constant (LC), which defines the relative rates of brain utilization of glucose and 2-deoxyglucose (2-DG), doubles to values > 1.0 under conditions of hypoglycemia. Since changes in the LC should be predictable given the kinetic parameters of blood-brain barrier (BBB) transport and brain phosphorylation of glucose and 2-DG, the present studies were designed to measure the necessary kinetic parameters. The carotid injection technique was used to determine cerebral blood flow and the Km , Vmax, and K D of glucose and 2-DG transport through the BBB in seven brain regions in rats anesthetized with 50 mg/kg i.p. pentobarbital. Regional glucose transport through the BBB was characterized by an average Km = 6.3 m m , average Vmax = 0.53 μmol min−1g−1, and average K D= 0.022 ml min−1g−1. The nonsaturable route of transport of glucose represented on the average 40% of the total glucose influx into brain regions at an arterial glucose concentration of 10 m m . In addition, the rate constants of phosphorylation of glucose and 2-DG were measured for each region. Substitutions of the measured kinetic parameters for sugar transport and phosphorylation into equations defining the LC confirm the observation that the LC would be expected to vary under extreme conditions such as hypoglycemia and to exceed values of 1.0 under these conditions.  相似文献   

5.
Abstract.  The hydrogen cyanide-based interaction of a strongly cyanogenic plant, Passiflora capsularis , and larvae of two insect herbivores, a generalist ( Spodoptera frugiperda ) and a specialist ( Heliconius erato ), is examined in terms of the combined kinetics of the feeding process and the simultaneous hydrogen cyanide (HCN) liberation, as compared with the natural kinetics of hydrogen cyanide evolution by plant-leaf tissue. There are marked differences in acceptance of P. capsularis by third-instar larvae of specialist and generalist species. The former, H. erato , display a parsimonious ingestion rate of 0.74 ± 0.15 mg (fresh weight) min−1 comprising 18% active feeding time, whereas S. frugiperda larvae show a more erratic and restrained feeding involving 4% of the time at 0.45 ± 0.14 mg min−1. These S. frugiperda larvae ingest 124.4 ± 8.3 mg (fw) of the non-cyanogeneic Spinacia oleracea leaves in 24 h compared with only 74.7 ± 20.1 mg of P. capsularis in the same period. The total hydrogen cyanide released naturally from wild specimens of P. capsularis plants is in the range 326–3901 μg g−1. Hydrogen cyanide evolution from macerated P. capsularis leaves takes place along a hyperbolic function with time and initial velocities of cyanide evolution are a linear function of total hydrogen cyanide. When feeding on P. capsularis leaves, H. erato releases only a minor fraction relative to total hydrogen cyanide (0.09%) and to the anticipated cyanide from the initial velocity (7%). By contrast, S. frugiperda evokes 5.8-fold more than the anticipated hydrogen cyanide release from the plant. The findings are interpreted as diverging strategies by generalist and specialist insects in the utilization of hydrogen cyanide in cyanogenic plants.  相似文献   

6.
Abstract: In this study we have examined (1) the integrated function of the mitochondrial respiratory chain by polarographic measurements and (2) the activities of the respiratory chain complexes I, II–III, and IV as well as the ATP synthase (complex V) in free mitochondria and synaptosomes isolated from gerbil brain, after a 30-min period of graded cerebral ischaemia. These data have been correlated with cerebral blood flow (CBF) values as measured by the hydrogen clearance technique. Integrated functioning of the mitochondrial respiratory chain, using both NAD-linked and FAD-linked substrates, was initially affected at CBF values of ∼35 ml 100 g−1 min−1, and declined further as the CBF was reduced. The individual mitochondrial respiratory chain complexes, however, showed differences in sensitivity to graded cerebral ischaemia. Complex I activities decreased sharply at blood flows below ∼30 ml 100 g−1 min−1 (mitochondria and synaptosomes) and complex II–III activities decreased at blood flows below 20 ml 100 g−1 min−1 (mitochondria) and 35–30 ml 100 g−1 min−1 (synaptosomes). Activities declined further as CBF was reduced below these levels. Complex V activity was significantly affected only when the blood flow was reduced below 15–10 ml 100 g−1 min−1 (mitochondria and synaptosomes). In contrast, complex IV activity was unaffected by graded cerebral ischaemia, even at very low CBF levels.  相似文献   

7.
Mercury concentrations in the axial muscle tissue of most (243) of the 254 Anguilla australis and most (20) of the 27 A. reinhardtii collected from 30 sites in coastal rivers and lakes in Victoria, Australia, during 1975–78 were well below the Australian statutory health limit (0.5 μg g−1 wet weight). For A. australis the mean mercury concentration was 0.17 μg g−1 (±0.16 s.d. , range 0.01–1.60 μg g−1); for A. reinhardtii the values were 0.37 ± 0.23 μg g−1 (range 0.12–1.10 μg g−1). Statistical analyses showed that variation in mercury concentration due to total length accounted for only 13% of the total variation in A. australis and 2% in A. reinhardtii whereas locality accounted for 54 and 68%, respectively. Both species are thus considered suitable as indicators of mercury pollution.  相似文献   

8.
Abstract: Data are presented in support of the transport of (-)- d -3-hydroxybutyrate across the blood-brain barrier (BBB) being a carrier-mediated process. The kinetic parameters in 21-day-old pentobarbital-anaesthetized rats were Vmax 2.0 μmol.g−1.min−1, K m 29 m M , and K D 0.024 ml.g−1.min−1. The value for Vmax was the same as that for l -lactate and pyruvate transport in animals of the same age. The transport of all three substrates was sensitive to inhibition by low concentrations of either 2-oxo-3-methylbutanoate or 2-0x0-4-methylpentanoate, the 2-oxo acids that can accumulate in patients with maple-syrup-urine disease. The K m values for the 2-oxo acids were severalfold lower than the respective K m values. 2-oxo-3-phenylpropionate was a poor inhibitor. The relative affinities of the various monocarboxylic acids for the transport system of the BBB distinguished it from similar systems described in brain, heart, and liver mitochondria; human erythrocytes; and Ehrlich ascites-tumour cells.  相似文献   

9.
Abstract: We measured hexokinase (EC 2.7.1.1) activity in particulate and soluble fractions isolated from bullfrog ( Rana catesbeiana ) retinas. Seventy-three percent of the hexokinase (HK) activity was associated with the particulate fraction, 27% with the soluble fraction. Both HK fractions could phosphorylate fructose, glucose, 2-deoxy-d-glucose, and mannose, but not galactose. The K m for glucose was 0.14 m M , for 2-deoxy-d-glucose. 3.6 m M . With glucose as substrate, the V max for particulate HK was 125–148 μ M retina−1 min−1, for soluble HK, 37 μ M retina−1 min−1. Product inhibition of particulate HK activity by glucose 6-phosphate was marked, whereas 2-deoxy-d-glucose 6-phosphate did not inhibit the activity. Cyclic AMP stimulated the HK activity of both retinal fractions nearly twofold at concentrations of 0.2–0.8 m M; AMP was much less effective in this regard.  相似文献   

10.
Polysiphonia paniculata Montagne is an intertidal red alga known to produce large amounts of the compound dimethylsulfoniopropionate (DMSP). Conversion of this substrate into dimethylsulfide is accomplished in P, paniculata by an enzyme called DMSP lyase (dimethylpropiothetin dethiomethyla.se (4.4.1.3)). DMSP lyase has been purified and characterized from P. paniculata. Enzymie activity is found in two different proteins: the larger with a molecular weight of 9.26 ± 104 daltons and the smaller with a molecular weight of 3.65 ± 104 daltons. Specific activity of the enzyme is 526 μmols min−1mg−1 for the smaller protein a nd 263 μmols min −1 mg−1 for the la rger protein. The Michaelis-Menten constant (Km) is 72.8 μM ± 17.15 and the vmax is 1.62 μmols min−1± 0.928 for the 92.6-kDa protein. The p1 of the larger protein is 5.8 and 5.9 for the smaller protein. Interaction with cysteine protease inhibitors L-trans-epoxysuccinyl-leucylamido (4-guanidino)-butane, dithiobis-(2-nitrobenzoate), or N -ethylmaleimide inactivated enzyme activity. The presence of either magnesium or calcium with DMSP lyase enhanced activity al concentrations between 20 and 40 μM but had little effect above these levels. Addition of the divalent chelators ethylenebis(oxyethylenenitrilo) tetraacetic acid and ethylenediaminetetraacetate decreased activity of the enzyme, but activity was restored when either chelator was removed and magnesium or calcium was added to the enzyme .  相似文献   

11.
Juvenile bull trout Salvelinus confluentus exposed to continuous- or pulsed-DC electroshock exhibited rapid elevations in plasma cortisol and glucose, but plasma chloride did not change. In a 1-h experiment using 240 V at 1·4 A of 60-Hz pulsed DC (voltage gradient 0·81 V cm−1), which proved lethal, plasma cortisol and glucose rose significantly within 15 min of a 10-s electroshock. Plasma cortisol reached a peak level of 156 ± 18 ng ml−1 at 45 min and then decreased, whereas plasma glucose reached its highest level of 179 ± 7·5mg dl−1 at 1 h. In a 24-h experiment using lower dosages, plasma cortisol increased from 6·1-16 ng ml−1 to peak levels of 155–161 ng ml−1 in 1 h in response to a 10-s electroshock of continuous (130 V, 0·5 A, 1·45 V cm−1) or pulsed (120 V, 0·5 A, 60 Hz, 0·55 V cm−1) DC. Although plasma concentrations declined thereafter, levels remained above control values at 24 h. Plasma glucose was elevated from 60–65 to 120–134 mg dl−1 after 1h by both electroshock treatments and remained near or above those levels for the 24-h duration. Plasma cortisol and glucose levels were much higher in electroshocked bull trout at 1 h compared with those in fish 1 h after receiving a 30-s handling stressor (cortisol, 90 ± 12 ng ml−1; glucose, 82 ± 6·1 mg dl−1). The results indicate that both continuous and pulsed DC were more stressful to juvenile bull trout than handling and that recovery, at least for pulsed DC, may take longer than 24 h.  相似文献   

12.
Abstract— The kinetics of the uptake from blood to brain of pyruvate, lactate and glucose have been determined in rats of different ages. The carotid artery single injection technique was used in animals anaesthetized with pentobarbital. The rates of influx for each substrate were determined over a range of concentrations for the different age-groups. Data were analysed in terms of the Michaelis-Menten equation with a component to allow for non-saturable diffusion. Values are given for K m, V max and K d. In suckling rats (15-21 days) the V max values for both pyruvate and lactate were 2.0 μmol g−1 min−1. In 28-day-old rats the V max values had fallen to one-half and in adults they were less than one-tenth. K m, values were higher in the younger animals. The rate of glucose transport in suckling rats was half that of 28-day-old and adults although there was no difference with age in the K m values.
The results are discussed in relation to the net flux of these substrates in and out of brain during different stages of post-natal development.  相似文献   

13.
Regulation of DOPA Decarboxylase Activity in Brain of Living Rat   总被引:4,自引:1,他引:3  
Abstract: To test the hypothesis that l -DOPA decarboxylase (DDC) is a regulated enzyme in the synthesis of dopamine (DA), we developed a model of the cerebral uptake and metabolism of [3H]DOPA. The unidirectional blood-brain clearance of [3H]DOPA ( K D1) was 0.049 ml g−1 min−1. The relative DDC activity ( k D3) was 0.26 min−1 in striatum, 0.04 min−1 in hypothalamus, and 0.02 min−1 in hippocampus. In striatum, 3,4-[3H]dihydroxyphenylacetic acid ([3H]DOPAC) was formed from [3H]DA with a rate constant of 0.013 min−1, [3H]homovanillic acid ([3H]HVA) was formed from [3H]DOPAC at a rate constant of 0.020 min−1, and [3H]HVA was eliminated from brain at a rate constant of 0.037 min−1. Together, these rate constants predicted the ratios of endogenous DOPAC and HVA to DA in rat striatum. Pargyline, an inhibitor of DA catabolism, substantially reduced the contrast between striatum and cortex, in comparison with the contrast seen in autoradiograms of control rats. At 30 min and at 4 h after pargyline, k D3 was reduced by 50% in striatum and olfactory tubercle but was unaffected in hypothalamus, indicating that DDC activity is reduced in specific brain regions after monoamine oxidase inhibition. Thus, DDC activity may be a regulated step in the synthesis of DA.  相似文献   

14.
The circadian rhythm in growth of the red macroalga Porphyra umbilicalis (Linnaeus) J. Agardh was investigated under different spectral light conditions in laboratory-grown thalli. A free-running rhythm was observed in constant green or red light at irradiances of 2.5 to 20 μmol photons·m−2·s−1, whereas arhythmicity occurred in constant blue light at 6–20 μmol photons·m−2·s−1. The circadian oscillator controlling growth rhythmicity in Porphyra uses most of the visible sunlight spectrum and possibly multiple photoreceptors with a high sensitivity for blue light and a lower sensitivity for red light. This was inferred from three experimental results: (1) The free-running period, τ, of the growth rhythm decreased with increasing irradiance, from approximately 25 h at 2.5 μmol photons·m−2·s−1 to 22 h at 20 μmol photons·m−2·s−1 in red or green light, (2) Dark pulses of 3 h duration, interrupting otherwise continuous green or red light, caused advances during the subjective day and delays during the subjective night; the circadian oscillator in Porphyra can discriminate darkness from green or red light, and (3) Low-irradiance blue light pulses (2.5 μmol photons·m−2·s−1) shifted the growth rhythm in red light of higher irradiance (e.g. 10 μmol photons·m−2·s−1), and a strong, high amplitude, type 0 phase response curve was obtained that is usually observed with light pulses shifting a circadian rhythm in otherwise continuous darkness.  相似文献   

15.
16.
Aims:  To develop solid-state fermentation system (SSF) for hyper production of tylosin from a mutant γ-1 of Streptomyces fradiae NRRL-2702 and its parent strain.
Methods and Results:  Various agro-industrial wastes were screened to study their effect on tylosin production in SSF. Wheat bran as solid substrate gave the highest production of 2500 μg of tylosin g−1 substrate by mutant γ-1 against parent strain (300 μg tylosin g−1 substrate). The tylosin yield was further improved to 4500 μg g−1 substrate [70% moisture, 10% inoculum (v/w), pH 9·2, 30°C, supplemental lactose and sodium glutamate on day 9]. Wild-type strain displayed less production of tylosin (655 μg of tylosin g−1 substrate) in SSF even after optimization of process parameters.
Conclusion:  The study has shown that solid-state fermentation system significantly enhanced the tylosin yield by mutant γ-1.
Significance and Impact of the Study:  This study proved to be very useful and resulted in 6·87 ± 0·30-fold increase in tylosin yield by this mutant when compared to that of wild-type strain.  相似文献   

17.
Abstract: Choline uptake in Y79 human retinoblastoma cells occurs through two kinetically distinguishable processes. The high-affinity system shows little sodium or energy dependence, and it does not appear to be linked to acetyl CoA: choline O -acetyltransferase. When the cells are grown in a culture medium containing 10% fetal bovine serum, the high-affinity system has a K' m= 2.16 ± 0.13 μ m and V' max= 27.0 ± 2.9 pmol min−1 mg−1, whereas the low-affinity system has K' m= 20.4 ± 1.3 μ m and V' max= 402 ± 49 pmol min−1 mg−1. Under these conditions, the polyunsaturated fatty acid content of the cell membranes is relatively low. When the polyunsaturated fatty acid content of the microsomal membrane fraction was increased by supplementing the culture medium with linolenic or docosahexaenoic acids (n-3 polyunsaturated fatty acids) or arachidonic acid (n-6 polyunsaturated fatty acid), the K' m of the high-affinity choline transport system was reduced by 40–60%. The V' max also was reduced by 20–40%. Supplementation with oleic acid, the most prevalent monounsaturated fatty acid, did not affect either kinetic parameter. The results suggest that one functional effect of the high unsaturated fatty acid content of neural cell membranes is to facilitate the capacity of the high-affinity choline uptake system to transport low concentrations of choline. This effect appears to be specific for polyunsaturated fatty acids but not for a single type, for it is produced by members of both the n-3 and n-6 classes of polyunsaturated fatty acids.  相似文献   

18.
The respiration of coho salmon, Oncorhynchus kisutch , weighing between 15 and 50 g was measured at gradually declining oxygen levels and at temperatures ranging between 14 and 17°C. The maximum and minimum oxygen concentrations tested were 250 and 40 μmol L−1, respectively. Respiration rates were measured for 1 h periods before oxygen concentration was lowered by 12.5 or 25.0 μmol oxygen L−1. At the end of these endurance tests the oxygen level was returned to normoxic conditions and respiration rates were determined for the recovery period. Under normoxic conditions (> 200 μmol L−1) the respiration of coho levelled around 5.1 μmol g−1 wet weight h−1. At intermediate levels between 150 and 200 μmol oxygen L−1, the average rate increased to 5.8 μmol g−1 h−1, which could be attributed to higher spontaneous activity of the test animals. At low oxygen levels (< 150 μmol−1) average respiration rates dropped to values between 5.5 and 5.7 μmol g−1 h−1, reaching a minimum of 3.8 μmol g−1 h−1 at oxygen levels below 50 μmol Lμ. First mortality was observed in this range. After exposure to reduced oxygen levels the fish maintained a higher respiration rate when again exposed to normoxic oxygen levels above 200 μmol L−1. Increased respiration rates were observed for a recovery period of 6 h.  相似文献   

19.
Abstract In Methanothrix soehngenii acetate is first activated by an acetate thiokinase rather than a phosphotransacetylase. The specific activity of the acetate thiokinase was 5.29 μmol acetate activated min−1 mg−1 protein with a half maximum rate at 0.74 mM acetate and at 0.047 mM CoA. In cell-free extracts a CO-dehydrogenase activity was measured of 3.02 μmol min−1 mg−1 protein with a half maximum rate at 0.44 mM CO and at 0.18 mM methylviologen. NADP and NAD could not replace methylviologen. F420 showed only low activity as electron acceptor.  相似文献   

20.
Abstract— Uptake and release of glutamine were measured in primary cultures of astrocytes together with the activity of the phosphate activated glutaminase (EC 3.5.1.2). In contrast to previous findings of an effective, high affinity uptake of other amino acids (e.g. glutamate, GABA) no such uptake of glutamine was observed, though a saturable, concentrative uptake mechanism did exist (K m = 3.3 ± 0.5 m m ; V max= 50.2 ± 12.6 nmol ± min−1± mg−1). The phosphate activated glutaminase activity in the astrocytes (6.9 ± 0.9 nmol ± min−1± mg−1) was similar to the activity found in whole brain (5.4 ± 0.7 nmol ± min −l± mg−1), which may contrast with previous findings of a higher activity of the glutamine synthetase (EC 6.3.1.2) in astrocytes than in whole brain. The observations are compatible with the hypothesis of an in vivo flow of glutamate (and GABA) from neurons to astrocytes where it is taken up and metabolized, and a compensatory flow of glutamine towards neurons and away from astrocytes although the latter cell type may be more deeply involved in glutamine metabolism than envisaged in the hypothesis.  相似文献   

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