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1.
Spectrophotometry of hemoglobin: a comparison of dog and man   总被引:1,自引:0,他引:1  
1. The absorptivity at 540 nm of hemiglobincyanide (epsilon 540HiCN) from dog blood was determined on the basis of iron and found to be within the range formerly obtained for human hemoglobin. 2. Consequently, epsilon 540HiCN = 11.0, the established value for human hemoglobin, may be used for dog hemoglobin. 3. On this basis the absorption spectra of oxyhemoglobin, de-oxygenated hemoglobin, carboxyhemoglobin, hemiglobin (methemoglobin) and hemiglobincyanide were determined for dog hemoglobin. 4. No significant differences were found between dog and human hemoglobin, except that dog hemiglobin binds less OH- as reflected in a difference between the absorption spectra of dog and human hemiglobin at the same pH.  相似文献   

2.
The binding and inhibitory properties of 11 benzimidazoles for bovine brain tubulin were investigated. The effects of the benzimidazoles on the initial rates of microtubule polymerization were determined by a turbidimetric assay. The median inhibitory concentrations (I50) for nocodazole, oxibendazole, parbendazole, mebendazole and fenbendazole ranged from 1.97 · 10−6 to 6.32 · 10−6 M. Benomyl, cambendazole and carbendazim had I50 values from 5.83 · 10−5 to 9.01 · 10−5 M. Thiabendazole had an I50 value of 5.49 · 10−4 M. Inhibitor constants (Ki) were determined by the colchicine binding assay. Oxibendazole, fenbendazole, and cambendazole had Ki values of 3.20 · 10−5, 1.73 · 10−5 and 1.10 · 10−4 M, respectively. Oxibendazole and fenbendazole were competitive inhibitors of colchicine. In contrast, cambendazole was a noncompetitive inhibitor of colchicine. The ability of these benzimidazoles to inhibit microtubule polymerization and the mode of action for the anthelmintic benzimidazoles is discussed.  相似文献   

3.
The iron requirement of a cell line of Chinese hamster fibroblasts is met more efficiently by human transferrin than by bovine transferrin or conalbumin. One possible explanation is that the binding of these transferrins to the Chinese hamster V79 cells may differ. Binding studies now show that the affinity of V79 cells for human transferrin is about 40 times greater than for bovine transferrin. Conalbumin has no detectable affinity for the human transferrin binding sites. Human apotransferrin has approximately one-sixth the affinity for the transferrin binding sites. The binding constant for the relation of human transferrin with the V79 cell is about 2.3·1061· mole−1, and the approximate number of binding sites per cell is 9 · 105.  相似文献   

4.
In this study, the hydraulic conductivity (Lp), Me2SO permeability ( Me2SO), and the reflection coefficients (ς) and their activation energies were determined for Metaphase II (MII) mouse oocytes by exposing them to 1.5 M Me2SO at temperatures of 30, 20, 10, 3, 0, and −3°C. These data were then used to calculate the intracellular concentration of Me2SO at given temperatures. Individual oocytes were immobilized using a holding pipette in 5 μl of an isosmotic PBS solution and perfused with precooled or prewarmed 1.5 M Me2SO solutions. Oocyte images were video recorded. The cell volume changes were calculated from the measurement of the diameter of the oocytes, assuming a spherical shape. The initial volume of the oocytes in the isoosmotic solution was considered 100%, and relative changes in the volume of the oocytes after exposure to the Me2SO were plotted against time. Mean (means ± SEM) Lpvalues in the presence of Me2SO ( Me2SOp) at 30, 20, 10, 3, 0, and −3°C were determined to be 1.07 ± 0.03, 0.40 ± 0.02, 0.18 ± 0.01, 7.60 × 10−2± 0.60 × 10−2, 5.29 × 10−2± 0.40 × 10−2, and 3.69 × 10−2± 0.30 × 10−2μm/min/atm, respectively. The Me2SOvalues were 3.69 × 10−3± 0.3 × 10−3, 1.07 × 10−3± 0.1 × 10−3, 2.75 × 10−4± 0.15 × 10−4, 7.83 × 10−5± 0.50 × 10−5, 5.24 × 10−5± 0.50 × 10−5, and 3.69 × 10−5± 0.40 × 10−5cm/min, respectively. The ς values were 0.70 ± 0.03, 0.77 ± 0.04, 0.81 ± 0.06, 0.91 ± 0.05, 0.97 ± 0.03, and 1 ± 0.04, respectively. The estimated activation energies (Ea) for Me2SOp, Me2SO, and ς were 16.39, 23.24, and −1.75 Kcal/mol, respectively. These data may provide the fundamental basis for the development of more optimal cryopreservation protocols for MII mouse oocytes.  相似文献   

5.
Two processes of porphyrins in heterogeneous systems containing aqueous and membrane phases have been studied with hematoporphyrin and hematoporphyrin derivative: Dimerization equilibrium in the aqueous phases and porphyrin-membrane binding equilibrium using liposomes as models for biological membranes. The interrelationship of aqueous aggregations and membrane binding was probed and the porphyrin aggregation state in the membrane, at equilibrium, was assessed. Fluorimetric techniques were employed. The dimerization equilibrium constants, at neutral pH and 37°C were found to be 2.8 · 105 M−1 and 1.9 · 106 M−1 for hematoporphyrin and its derivative, respectively. Over a porphyrin concentration range going from monomer-dominant to dimer-dominant systems, we have found that only monomers are bound to the membrane. The respective monomer-liposome binding constants, found to be independent of the initial monomer/dimer distribution in the aqueous phase, were determined to be 1.6 · 103 M−1 and 4.1 · 103 M−1 at neutral pH and 37°C for hematoporphyrin and its derivative, respectively. The monomer-liposome interaction was found to perurb the initial monomer/dimer distribution in the aqueous phase, so that the monomers residing at equilibrium in the membrane originate from both monomers and dimers in the aqueous phase.  相似文献   

6.
Glycerol diffusional permeabilities through the cytoplasmic cell membrane of Dunaliella salina, the cell envelope of pig erythrocyte and egg phosphattidylcholine vesicles were measured by NMR spectroscopy employing the spin-echo method and nuclear T1 relaxation. The following permeability coefficients (P) and corresponding enthalpies of activation (ΔH) were determined for glycerol at 25°C: for phosphatidylcholine vesicles 5·10−6 cm/s and 11±2 kcal/mol; for pig erythrocytes 7·10−8 cm/s and 18±3 kcal/mol, respectively; for the cytoplasmic membrane of D. salina the permeability at 17°C was found to be exceptionally low and only a lower limit (P<5·10−11cm/s) could be calculated. At temperatures above 50°C a change in membrane permeability occurred leading to rapid leakage of glycerol accompanied by cell death. The data reinforce the notion that the cytoplasmic membrane of Dunaliella represents a genuine anomaly in its exceptional low permeability to glycerol.  相似文献   

7.
Noradrenalin (8 · 10−6 M) and adrenalin (6 · 10−6 and 6 · 10−7 M) were found to cause marked stimulation of short-circuit current (S.C.C.) in isolated toad bladder, but isoprenalin (8 · 10−7 M) was found to be without effect. The percentage rise in S.C.C. due to noradrenalin was found to be inversely proportional to the initial S.C.C. or total conductance of the bladder. Again in the case of noradrenalin the rise in S.C.C. was almost completely abolished by α-adrenergic blockade but not by β-blockade. This rise in S.C.C. was found not to be significantly different from the rise in net Na+ flux. Bidirectional Cl fluxes were estimated using 82Br as a companion radionuclide to 36Cl. No significant net Cl flux was apparent, either before or after addition of any of the three catecholamines tested. However, in some cases the unidirectional Cl fluxes rose markedly following addition of noradrenalin or of adrenalin and this change was not reflected in a change in total conductance. This anomaly was noted to occur in bladders whose initial conductance was of the order of 0.5 kΩ−1 · cm−2 or greater. The evidence presented suggests that two actions of catecholamines on ion transport in toad bladder are (a) to increase Na+ transport via stimulation of α-adrenergic sites and (b) at the concentrations tested to cause an increase in passive Cl permeability in bladders whose initial conductance is high.  相似文献   

8.
1. The fat mouse Steatomys pratensis natalensis (mean body mass 37.4±0.43 (se)) has a low euthermic body temperature Tb=30.1–33.8 °C and a low basal metabolic rate (BMR)=0.50 ml O2 g−1 h−1.
2. Below an ambient temperature (Ta)=15 °C, the mice were hypothermic.
3. The lowest survivable Ta=10 °C.
4. Torpor is efficient in conserving energy between Ta=15–30 °C, below Ta=15 °C, the mice arouse.
5. Euthermic and torpid mice were hyperthermic at Ta=35 °C.
6. Thermal conductance was 0.159 ml O2 g−1 h−1 °C−1, 98.8% of the expected value.
7. Non-shivering thermogenesis (NST) was 2.196 ml O2 g−1 h−1 (3.69×BMR).
8. Maximal oxygen consumption, however, was 3.83 ml O2 g−1 h−1 (6.44×BMR), indicating that other methods of heat production are additive.
9. Because fat mice conserve energy by torpor only between Ta=15–30 °C, we suggest that torpor may be a more important mechanism for surviving food shortages than for surviving cold weather.
Keywords: Steatomys pratensis natalensis; Metabolism; Torpor; Fat mouse  相似文献   

9.
Dispersed acini from dog pancreas were used to examine the ability of dopamine to increase cyclic AMP cellular content and the binding of [3H]dopamine. Cyclic AMP accumulation caused by dopamine was detected at 1·10−8 M and was half-maximal at 7.9±3.4·10−7M. The increase at 1·10−5 M, (7.5-fold) was equal to the half-maximal increase caused by secretin at 1·10−9 M. Haloperidol, a dopaminergic receptor antagonist inhibited cyclic AMP accumulation caused by dopamine. The IC50 value for haloperidol, calculated from the inhibition of cyclic AMP increase caused by 1·10−5 M dopamine was 2.3±0.9·10−6M. Haloperidol did not alter basal or secretin-stimulated cyclic AMP content. [3H]Dopamine binding was studied on the same batch of cells as cyclic AMP accumulation. At 37°C, it was rapid, reversible, saturable and stereospecific. The Kd value for high affinity binding sites was 0.43±0.1·10−7M and 4.7±1.6·10−7M for low affinity binding sites. The concentration of drugs necessary to inhibit specific binding of dopamine by 50% was 1.2±0.4·10/t-7M noradrenaline, 2·10/t-7 M epinine, 4.1±1.8·10/t-6M fluphenazine, 8.0±1.6·10/t-6M haloperidol, 4.2±1.2·10−6Mcis-flupenthixol, 2.7±0.4·10−5Mtrans-flupenthixol, >1·10−5M apomorphine, sulpiride, naloxone and isoproterenol.  相似文献   

10.
Thyroid hormone (T3) has been demonstrated to inhibit the action of aldosterone on sodium transport in toad urinary bladder and rat kidney. We have exammined the effect of T3 on aldosterone action and specific nuclear binding in cultured epithelial cells derived from toad urinary bladder. In cell line TB6-C, addition of 5·10−8 M T3 to culture media for up to 3 days results in no change in short-circuit current or transepithelial resistance. This concentration of T3 completely inhibits the maximal increase in short-circuit current in response to 1·10−7 M aldosterone. The inhibition can be demonstrated with 18 h preincubation or with simultaneous addition of T3 and aldosterone. The half-maximal concentration for the inhibition of the aldosterone effect is approx. 5·10−9 M T3. T3 has no effect on cyclic AMP-stimulated short-circuit current in these cells. The effect of T3 on nuclear binding of [3H]aldosterone was examined using a filtration assay with data analysis by at least-squares curve-fitting program. Best fit was obtained with a model for two binding sites. The dissociation constants for the binding were Kd1 = (0.82 ± 0.36)·10−10 M and Kd2 = (3.2±0.60)·10−8 M.The half-maximal concentration for aldosterone-stimulated sodium transport in these cells is approx. 1·10−8 M. Analysis of nuclear aldosterone binding in cells preincubated for 18 h with 5·10−8 M T3 showed a Kd1 = (0.15 ± 0.10)·10−10 M and Kd2 = (3.5 ± 0.10)·10−8 M. We conclude that T3 i action of aldosterone on sodium transport at a site after receptor binding in the nucleus.  相似文献   

11.
Studies on mixed mass cultivation of Anabaena spp. on a large scale (5170m2) were conducted continuously for 3 years. Under the continental monsoon climate in northern subtropics (30°N, 115°E), 7–11 g dry weight m−2 day−1 of microalgal biomass on average was harvested in simple plastic greenhouses in the effective growth days during the warmer seasons. The maximum productivity was 22 g m−2 day−1 in the middle of summer. Observations on the productive properties of strains of Anabaena spp. indicated that they were different from and could compensate for each other in their productivities and adaptations to the seasonal changes. With different lining materials (PVC sheets, concrete, sand and soil) in the culture ponds, no significant variation of productivity was found, but bubbling with biogas in the middle of the day and the application of some growth regulating substances (2,4-D, NaHSO3 and extracts of oyster mushroom spawn) was able to improve the production. The cost of microalgal biomass in this way was around 0·75–1·0 US dollar(s) per kilogram.  相似文献   

12.
Active transport of Cl accounts for 90% of the short-circuit current (s.c.c.) in the isolated frog cornea. 1·10−5 M furosemide produced a 50% reversible inhibition of this s.c.c. 1·10−4 M ethacrynic acid reduced the corneal s.c.c. to 32% of the control. In the isolated frog skin epithelium furosemide had no effect on the s.c. at a concentration of 1·10−4 M and a small stimulation at a concentration of 1·10−3 M. The furosemide inhibitory effects seems to be specific for Cl, as it also inhibits Cl transport in the ascending limb of the loop of Henle (Burg, M.B. (1972) Proc. 5th Int. Congr. Nephrol., p. 50, Abstr.).  相似文献   

13.
A hydrogen peroxide permselective membrane with asymmetric structure was prepared and -glucose oxidase (EC 1.1.3.4) was immobilized onto the porous layer. The activity of the immobilized -glucose oxidase membrane was 0.34 units cm−2 and the activity yield was 6.8% of that of the native enzyme. Optimum pH, optimum temperature, pH stability and temperature stability were found to be pH 5.0, 30–40°C, pH 4.0–7.0 and below 55°C, respectively. The apparent Michaelis constant of the immobilized -glucose oxidase membrane was 1.6 × 10−3 mol l−1 and that of free enzyme was 4.8 × 10−2 mol l−1. An enzyme electrode was constructed by combination of a hydrogen peroxide electrode with the immobilized -glucose oxidase membrane. The enzyme electrode responded linearly to -glucose over the concentration 0–1000 mg dl−1 within 10 s. When the enzyme electrode was applied to the determination of -glucose in human serum, within day precision (CV) was 1.29% for -glucose concentration with a mean value of 106.8 mg dl−1. The correlation coefficient between the enzyme electrode method and the conventional colorimetric method using a free enzyme was 0.984. The immobilized -glucose oxidase membrane was sufficiently stable to perform 1000 assays (2 to 4 weeks operation) for the determination of -glucose in human whole blood. The dried membrane retained 77% of its initial activity after storage at 4°C for 16 months.  相似文献   

14.
We measured Na+/K+ ATPase activity in homogenates of gill tissue prepared from field caught, winter and summer acclimatized yellow perch, Perca flavescens. Water temperatures were 2–4°C in winter and 19–22°C in summer. Na+/K+ ATPase activity was measured at 8, 17, 25, and 37°C. Vmax values for winter fish increased from 0.48±0.07 μmol P mg−1 protein h−1 at 8°C to 7.21±0.79 μmol P mg−1 protein h−1 at 37°C. In summer fish it ranged from 0.46±0.08 (8°C) to 3.86±0.50 (37°C) μmol P mg−1 protein h−1. The Km for ATP and for Na+ at 8°C was ≈1.6 and 10 mM, respectively and did not vary significantly with assay temperature in homogenates from summer fish. The activation energy for Na+/K+ ATPase from summer fish was 10 309 (μmol P mg−1 h−1) K−1. In winter fish, the Km for ATP and Na+ increased from 0.59±0.08 mM and 9.56±1.18 mM at 8°C to 1.49±0.11 and 17.88±2.64 mM at 17°C. The Km values for ATP and Na did not vary from 17 to 37°C. A single activation energy could not be calculated for Na/K ATPase from winter fish. The observed differences in enzyme activities and affinities could be due to seasonal changes in membrane lipids, differences in the amount of enzyme, or changes in isozyme expression.  相似文献   

15.
The binding of biliverdin-IXα by human albumin and serum was quantitated, using three different binding techniques, to study the effects of biliverdin on bilirubin-albumin binding. The apparent equilibrium association constants (K ± SD) and binding capacities (n) of defatted albumin, pooled adult sera, and pooled umbilical cord sera for biliverdin are: K = 1.3 ± 2 × 106 −1, n = 1.00; K = 13.0 ± 3 × 106 −1, n = 0.90; and K = 6.8 ± 0.1 × 106 −1, n = 0.85, respectively. Although bilirubin binds at more than one albumin site, competitive studies showed that biliverdin binds only at the primary (highest affinity) bilirubin site. Sulfisoxazole, previously thought to compete with bilirubin for the primary binding site, was found to displace bilirubin from both primary and secondary bilirubin binding sites. Biliverdin, because of its specific binding and spectral characteristics, could be a useful probe for determining the capacity of the primary bilirubin-albumin binding site.  相似文献   

16.
1. 1. The Michaelis-Menten parameters of labelled d-glucose exit from human erythrocytes at 2°C into external solution containing 50 mM d-galactose were obtained. The Km is 3.4 ± 0.4 mM, V 17.3 ± 1.4 mmol · 1−1 cell water · min−1 for this infinite-trans exit procedure.
2. 2. The kinetic parameters of equilibrium exchange of d-glucose at 2°C are Km = 25 ± 3.4 mM, V 30 ± 4.1 mmol · 1−1 cell water · min−1.
3. 3. The Km for net exit of d-glucose into solutions containing zero sugar is 15.8 ± 1.7 mM, V 9.3 ± 3.3 mol 9.3 ± 3.3 mol · 1−1 cell water · min−1.
4. 4. This experimental evidence corroborates the previous finding of Hankin, B.L., Lieb, W.R. and Stein, W.D. [(1972) Biochim. Biophys. Acta 255, 126–132] that there are sites with both high and low operational affinities for d-glucose at the inner surface of the human erythrocyte membrane. This result is inconsistent with current asymmetric carrier models of sugar transport.
Keywords: d-Glucose transport; Asymmetric carrier; Pore kinetics; (Erythrocyte)  相似文献   

17.
The technique of affinity chromatography with the curarizing neurotoxins of Naja naja venom has been employed to extract nicotinic acetylcholine receptors from the brain tissues of mouse and hog. Both carbochol and hexamethonium were used as linear or step gradients to elute the receptor and its properties were investigated in lipid bilayer membranes. Of particular interest is the observation that discrete quanta of conductance could be observed across an NaCl gradient of 1.0:0.1 M. By switching the voltage-clamp across the bilayer between a positive and negative 80 mV, the separate Na+ and Cl conductances of these quanta could be estimated and the following conductances of the smallest discrete quanta were observed: 3.7 · 10−11 Ω−1 (Na+) and 5.9 · 10−11 Ω−1 (Cl) for mouse brain receptors; 3.8 · 10−11 Ω−1 (Na+) and 4.7 · 10−11 Ω−1 (Cl) for hog brain receptors. Large aggregates of receptors appeared to activate and deactivate as multiples of a basic conductance size, although there is evidence that they may not represent the actual gating of ion channels. A “background noise” that is not within the temporal capability of the recording system is also present at an intensity that seems to parallel the number of activated receptors, and in view of recent electrophysiological evidence that the relaxation lifetime of the open channel state is of a millisecond duration, it may be that this “noise” actually represent the channel gating.  相似文献   

18.
Electrorotation of single platelets was compared with [14C]serotonin release, aggregation and electron microscopy. Activation of washed and degranulated platelets was induced by thrombin, arachidonic acid, collagen, adrenaline, platelet activation factor (PAF (PA), ADP and A23187. A strong correlation between electrorotation decrease and serotonin release was found. Electrorotation did not correlate with aggregation. It was concluded that an increase of the specific conductivity of the platelet membrane by three orders of magnitude (approx. 1.0–10−7 S · m−1 to 1.0·10−4 S · m−1) upon activation was responsible for the observed decrease of anti-field rotation and the shift of the first characteristic frequency towards higher values. Electrorotation allowed for time-dependent measurements of activation. Characteristic activation times in the order of minutes were found. There was the following sequence of activators classified by increasing activation time constants: A23187 was the fastest followed by thrombin, collagen, PAF, arachidonic acid, adrenaline, and ADP.  相似文献   

19.
Human α1-antitrypsin (AAT) was produced in the recombinant yeast Saccharomyces cerevisiae ATCC 20699 grown in batch and fed-batch culture. The final biomass concentration and antitrypsin concentration attained were 55 g·L−1 and 1.23 g·L−1, respectively, in the fed-batch. The maximum productivities of biomass and antitrypsin were 1.6 and > 0.04 g L−1h−1, respectively, or substantially greater than the highest productivity values reported in the past. For recovering the antitrypsin, the cell slurry was concentrated 4-fold (231 g·L−1 biomass, 122 min of processing) by cross-flow microfiltration and the cells were disrupted by bead milling (3 passes of 3 min total retention time). The cell homogenate was treated with aluminum chloride or PBS (pH 7) to aid separation of the cell debris by flocculation and sedimentation. The clarified cell homogenate was subjected to ammonium sulfate fractionation to precipitate the recombinant antitrypsin. The AAT precipitated at 45–75% saturation of ammonium sulfate, depending on the age of the homogenate. The crude AAT in the homogenate degraded at room temperature (25°C), with a zero order deactivation rate of 1.815 × 10−3 ± 3.43 × 10−4 g AAT L−1h−1.  相似文献   

20.
Clones 02 and 4430 of Tradescantia were tested in field, greenhouse and controlled environment chambers as monitors for the potentially hazardous UV-B irradiation increase that could result from stratospheric ozone decrease. In addition to about 16 hr of solar emissions at about 2100 micro-einsteins·m−2·s−1 (400–700 nm) and 15 hr at about 1800 micro-einsteins·m−2·s−1 in the field and greenhouse, respectively, plants were given 7 hr of supplemental UV-B irradiation per day for 27 days. After the first 7 days of UV-B irradiation exposure, cumulative data were recorded for 20 days. Cuttings of Tradescantia plants in controlled-environment, exposed to 16 hr of simulated solar emission of about 800 micro-einsteins·m−2·s−1 (400–700 nm), were also exposed to 10 hr of supplemental UV-B irradiation per day for 1 or 2 days. All plants were checked for somatic aberrations (color changes in the flower petals and stamen hairs), number of hairs per stamen, and cells per hair. Pollen germination and pollen tube growth were noted after a 90-min UV-B irradiation period.Somatic aberrations occurred infrequently in the petals and were judged unreliable criteria for use in monitoring enhanced UV-B irradiation environments. The number of aberrant events within stamen hairs, however, was significantly increased by the UV-B irradiation treatments. while pollen germination and pollen tube growth were significantly reduced. These data indicate that color changes in stamen hairs and pollen viability are useful criteria for monitoring UV-B irradiation changes.  相似文献   

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