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1.
The tetrameric heart isozyme of lactate dehydrogenase (H4) is modified by p-chloromercuribenzoate (PCMB) to produce the inactive tetramer and then hybridized with native tetrameric muscle isozyme (M4). The hybrid mixture was isolated by polyacrylamide gel electrophoresis (PAGE) and then stained for enzyme activity and with Coomassie brilliant blue. Only three bands were found on the gels in either case. The hybrid enzymes as isolated by PAGE have half the specific activity of the native muscle enzyme. The electrophoresis properties of HM3 are very similar to those of HM3, while the electrophoresis properties of are very similar to those of H2M2. The above results strongly suggest that the tetramer having enzymatic activity contains at least two native subunits, and the di-subunit in the tetrameric enzyme is the minimal functional unit.  相似文献   

2.
The cyanobacterium Spirulina platensis was used to verify the possibility of employing microalgal biomass to reduce the contents of nitrate and phosphate in wastewaters. Batch tests were carried out in 0.5 dm3 Erlenmeyer flasks under conditions of light limitation (40 mol quanta m–2 s–1) at a starting biomass level of 0.50 g/dm3 and varying temperature in the range 23–40°C. In this way, the best temperature for the growth of this microalga (30°C) was determined and the related thermodynamic parameters were estimated. All removed nitrate was used for biomass growth (biotic removal), whereas phosphate appeared to be removed mainly by chemical precipitation (abiotic removal). The best results in terms of specific and volumetric growth rates ( =0.044 day–1, Q x =33.2 mg dm–3 day–1) as well as volumetric rate and final yield of nitrogen removal ( =3.26 mg dm–3 day–1, =0.739) were obtained at 30°C, whereas phosphorus was more effectively removed at a lower temperature. In order to simulate full-scale studies, batch tests of nitrate and phosphate removal were also performed in 5.0 dm3 vessels (mini-ponds) at the optimum temperature (30°C) but increasing the photon fluence rate to 80 mol quanta m–2 s–1 and varying the initial biomass concentration from 0.25 to 0.86 g/dm3. These additional tests demonstrated that an increase in the inoculum level up to 0.75 g/dm3 enhanced both NO3 and PO4 3– removal, confirming a strict dependence of these processes on biomass activity. In addition, the larger surface area of the ponds and the higher light intensity improved removal yields and kinetics compared to the flasks, particularly concerning phosphorus removal ( =0.032–0.050 day–1, Q x =34.7–42.4 mg dm–3 day–1, =3.24–4.06 mg dm–3 day–1, =0.750–0.879, =0.312–0.623 mg dm–3 day–1, and =0.224–0.440).  相似文献   

3.
Midstalk rot caused by Sclerotinia sclerotiorum is an important disease of sunflower in its main areas of cultivation. The objectives of this study were to (1) verify quantitative trait loci (QTL) for midstalk-rot resistance found in F3 families of the NDBLOSsel × CM625 population in recombinant inbred lines (RIL) derived from the same cross; (2) re-estimate their position and genetic effects; (3) draw inferences about the predictive quality of QTL for midstalk-rot resistance identified in the F3 families as compared to those in the RIL. Phenotypic data for three resistance (leaf lesion, stem lesion, and speed of fungal growth) and two morphological traits (leaf length and leaf length with petiole) were obtained from 317 RIL following artificial infection in field experiments across two environments. For genotyping the 248 RIL, we selected 41 simple sequence repeat (SSR) markers based on their association with QTL for Sclerotinia midstalk-rot resistance in an earlier study. The resistance traits showed intermediate to high heritabilities and were significantly correlated with each other Genotypic correlations between F3 families and the RIL were highly significant and ranged between 0.50 for leaf length and 0.64 for stem lesion. For stem lesion, two genomic regions on linkage group (LG) 8 and LG16 explaining 26.5% of the genotypic variance for Sclerotinia midstalk-rot resistance were consistent across generations. For this trait, the genotypic correlation between the observed performance and its prediction based on QTL positions and effects in F3 families was surprisingly high The genetic effects and predictive quality of these two QTL are promising for application in marker-assisted selection to Sclerotinia midstalk-rot resistance.  相似文献   

4.
Regional chemical potential values- R have been obtained with the use of nuclear reactivity indices. Perturbational formulae use values of reactivity indices of isolated molecular fragments. The changes of the parameters (NR,{ Qi }i R) within each fragment determine the value of the regional chemical potential after a chemical reaction. The computational scheme has been tested numerically along the chemical reaction path. We have studied a set of chemical reactions to obtain regional chemical potentials ( ) and regional transfer potentials ( ) for transition states of the following chemical reactions: HF+CO=HFCO, HCl+CO=HClCO, HF+SiO=HFSiO and HF+GeO=HFGeO. The results are reasonable and encouraging. Values of these indices show the possible reactivity directions of the transition states examined.  相似文献   

5.
Summary The Lipari-Szabo dynamical formalism is extended by setting the time constants of the Lorentzian terms to and . This analysis is compared to the earlier proposed three-parameter extended model free formalism with regard to the range of equivalence and the advantages of the simplified two-parameter (S inff sup2 ,S infH sup2 ) and (S inff sup2 ,S infN sup2 ) representations. Spectral density components are calculated and compared to those obtained from the spectral density analysis formalism. Protein relaxation data, commonly analyzed in terms of the two-parameter representation, may correspond to a dynamically heterogeneous behaviour that is more appropriately represented in terms of a fast limit order parameter and a second, lower frequency order parameter.  相似文献   

6.
We present long-term nutrient data on the Changjiang River (Yangtze River) at six hydrological stations and eight principal tributaries during the period 1958–1985. Three patterns of temporal changes were observed in nitrate and nitrite : minimal variations in the upper catchment area, rapid increases in the middle watershed towards the end of the 1970s, and a gradual increase in the lower drainage basin. Prior to the 1970s, the level of throughout the Changjiang River system remained fairly constant. In the 1980s, however, this changed, with the lowest values in the upper Changjiang changing rapidly to the highest in the middle reaches and then declining slowly but steadily in the lower courses. Compared to and ammonium and soluble reactive phosphorus (SRP) showed smaller increases or no long-term variations, while dissolved silica (DSi) concentration generally decreased at most stations. These three patterns of and changes in the Changjiang River system were reflective of the difference in chemical fertilizer use and landscape features (e.g., slope, soil type and water body area) of the drainage basins of the primary tributaries. The decreases in DSi were most likely attributed to a reduction in suspended sediment loading due to dam constructions and increasing diatom consumption. The increase in and with a reduction in DSi concentrations in the Changjiang River could have significant effects on the stoichiometric balance of nutrients delivered to the East China Sea and the ecosystem in this dynamic region.  相似文献   

7.
The dynamics of the backbone NH bonds of protein HU from Bacillus stearothermophilus (HUBst) have been characterized using measurements of cross-relaxation, longitudinal and transverse relaxation rates at 11.7, 14.1 and 17.6 T. Linear regression of the values with the squared Larmor frequency N 2 has revealed global exchange processes, which contributed on the order of 0.5–5.0 s-1to the transverse relaxation rate. Subsequently, the experimental values were corrected for these exchange contributions. A reduced spectral density mapping procedure has been employed with the experimental relaxation rates and seven values of the spectral density function J() have been extracted. These spectral densities have been fitted within the framework of the model-free approach. The densities agree well with an axially symmetric rotational diffusion tensor with a diffusion anisotropy D_/D_ of 1.15, indicating that the flexible arms of HUBst do not significantly contribute to the rotational diffusion. The overall correlation time is 8.9 ± 0.6 ns/rad. The fast internal motions of most of the NH bonds in the core display order parameters ranging between 0.74 and 0.83 and internal correlation times between 1 and 20 ps. For the residues in the DNA-binding -arms, an extended version of the model function has been used. The slow internal motions show correlation times of 1–2 ns. The concomitant order parameters (0.3–0.6) are lower than those observed on the fast time scale, indicating that the flexibility of the -arms is mainly determined by the slower internal motions. A substantial decrease of the generalized order parameters in the -arms starting at residues Arg55 and Ser74, opposite on both strands of the -ribbon arms, has been explained as a hinge motion. A comparison of the order parameters for free and DNA-bound protein has demonstrated that the slow hinge motions largely disappear when HU binds DNA.  相似文献   

8.
Summary A set of three-dimensional triple-resonance experiments is described which provide , , and coupling constants. The pulse sequences generate E.COSY-like multiplet patterns and comprise a magnetization transfer from the amide proton to the α-proton or vice versa via the directly bound heteronuclei. For residues with the 1Hα spin resonating close to the H2O signal, a modified HNCA experiment can be employed to measure the vicinal 1HN,1Hα couplings. Ambiguities associated with the conversion of values into ϕ-angle constraints for protein structure determination can be resolved with the knowledge of the heteronuclear 3J-couplings. In favourable cases, stereospecific assignments of glycine α-protons can be obtained by employing the experiments described here in combination with NOE data. The methods are applied to flavodoxin from Desulfovibrio vulgaris.  相似文献   

9.
Summary The energy requirements of Adélie penguin (Pygoscelis adeliae) chicks were analysed with respect to body mass (W, 0.145–3.35 kg, n=36) and various forms of activity (lying, standing, minor activity, locomotion, walking on a treadmill). Direct respirometry was used to measure O2 consumption ( ) and CO2 production. Heart rate (HR, bpm) was recorded from the ECG obtained by both externally attached electrodes and implantable HR-transmitters. The parameters measured were not affected by hand-rearing of the chicks or by implanting transmitters. HR measured in the laboratory and in the field were comparable. Oxygen uptake ranged from in lying chicks to at maximal activity, RQ=0.76. Metabolic rate in small wild chicks (0.14–0.38 kg) was not affected by time of day, nor was their feeding frequency in the colony (Dec 20–21). Regressions of HR on were highly significant (p< 0.0001) in transmitter implanted chicks (n=4), and two relationships are proposed for the pooled data, one for minor activities ( ), and one for walking ( ). Oxygen consumption, mass of the chick (2–3 kg), and duration of walking (T, s) were related as , whereas mass-specific O2 consumption was related to walking speed (S, m·s-1) as .Abbreviations bpm beats per minute - D distance walked (m) - ECG electrocardiogram - HR heart rate (bpm) - ns number of steps - RQ respiratory quotient - S walking speed (m·s-1) - T time walked (s) - W body mass (kg)  相似文献   

10.
Both trans- and cis-[PtCl2(NH3)(L)] compounds have been synthesized, L representing either the imino ether HN=C(OMe)Me having a Z or E configuration at the C=N double bond, or the cyclic ligands and (compounds 14 for trans geometry and 58 for cis geometry, respectively). The cyclic ligands mimic the imino ether ligands but, differently from imino ethers, cannot undergo change of configuration. In a panel of human tumor cells, trans compounds inhibit growth much more than transplatin. Moreover, compound 1 in most cases is less active than 2, and 1 and 2 are less active than 3 and 4, respectively. For cis compounds with imino ethers, the activity is reduced (5) or unaffected (6) with respect to cisplatin. Moreover, unlike trans compounds, substitution of cyclic ligands (7, 8) for imino ethers (5, 6) generally decreases the activity. This determines, for compounds with cyclic ligands, an unusual inversion of the cis geometry requirement for activity of platinum(II) species. Importantly, 14 and 58 partially circumvent the multifocal cisplatin resistance of A2780cisR cells, and 14 also overcome resistance from reduced uptake of 41McisR cells. DNA interaction regioselectivity of 14 and 58 is not substantially modified with respect to transplatin and cisplatin. However, both imino ethers and cyclic ligands slow down the DNA interstrand cross-link reaction, (E)-HN=C(OMe)Me and decreasing also its extent. Therefore, DNA interaction of 14 and 58 appears to be characterized by persistent monoadducts (14), and by monoadducts and/or intrastrand cross-links structurally different from those of cisplatin (58). This study demonstrates that ligand configuration modulates the activity of both trans and cis compounds, and supports the development of platinum drugs based on their coordination chemistry to combat cisplatin resistance.F.P. Intini and A. Boccarelli contributed equally to this work  相似文献   

11.
The recently defined versus straight-line plots for L = pyridine-type (PyN) and ortho-aminopyridine-type (oPyN) ligands now allow the evaluation in a quantitative manner of the stability of the 1:1 complexes formed between cytidine (Cyd) and Ca2+, Mg2+, Mn2+, Co2+, Ni2+, Cu2+, Zn2+ or Cd2+ (M2+); the corresponding stability constants, , including the acidity constant, , for the deprotonation of the (N3)H+ site had been determined previously under exactly the same conditions as the mentioned plots. Since the stabilities of the M(PyN)2+ and M(oPyN)2+ complexes of Ca2+ and Mg2+ are practically identical, it is concluded that complex formation occurs in an outer-sphere manner, and this is in accord with the fact that in the pKa range 3–7 metal ion binding is independent of or . Ca(Cyd)2+ and Mg(Cyd)2+ are more stable than the corresponding (outer-sphere) M(PyN)2+ complexes and this means that the C2 carbonyl group of Cyd must participate, next to N3 which is most likely outer-sphere, in metal ion binding, leading thus to chelates; these have formation degrees of about 50% and 35%, respectively. Co(Cyd)2+ and Ni(Cyd)2+ show no increased stability based on the hence, the (C2)O group does not participate in metal ion binding, but the inner-sphere coordination to N3 is strongly inhibited by the (C4)NH2 group. In the M(Cyd)2+ complexes of Mn2+, Cu2+, Zn2+ and Cd2+, this inhibiting effect on M2+ binding at N3 is partially compensated by participation of the (C2)O group in complex formation and the corresponding chelates have formation degrees between about 30% (Zn2+) and 83% (Cu2+). The different structures of the mentioned chelates are discussed in relation to available crystal structure analyses. (1) There is evidence (crystal structure studies: Cu2+, Zn2+, Cd2+) that four-membered rings form, i.e. there is a strong M2+ bond to N3 and a weak one to (C2)O. (2) By hydrogen bond formation to (C2)O of a metal ion-bound water molecule, six-membered rings, so-called semichelates, may form. (3) For Ca2+ and Mg2+, and possibly Mn2+, and their Cyd complexes, six-membered chelates are also likely with (C2)O being inner-sphere (crystal structure) and N3 outer-sphere. (4) Finally, for these metal ions also complexes with a sole outer-sphere interaction may occur. All these types of chelates are expected to be in equilibrium with each other in solution, but, depending on the metal ion, either the one or the other form will dominate. Clearly, the cytidine residue is an ambivalent binding site which adjusts well to the requirements of the metal ion to be bound and this observation is of relevance for single-stranded nucleic acids and their interactions with metal ions. In addition, the antisyn energy barrier has been estimated as being in the order of 6–7.5 kJ/mol for cytidine derivatives in aqueous solution at 25 °C.Abbreviations ADP3– adenosine 5-diphosphate - AMP2– adenosine 5-monophosphate - ATP4– adenosine 5-triphosphate - CDP3– cytidine 5-diphosphate - cl closed - CMP2– cytidine 5-monophosphate - 3-CMP2– cytidine 3-monophosphate - CTP4– cytidine 5-triphosphate - Cyd cytidine - DNA deoxyribonucleic acid - I ionic strength - Ka acidity constant - KI intramolecular equilibrium constant - L general ligand - M2+ general divalent metal ion - NTP4– nucleoside 5-triphosphate - op open - oPyN ortho-aminopyridine-type ligand - PyN pyridine-type ligand - t-RNA transfer ribonucleic acid - Tu tubercidin (7-deazaadenosine)In honor of Professor Liang-Nian Ji on the occasion of his 70th birthday in friendship and with best wishes.  相似文献   

12.
Polysulphide was formed according to reaction (1), when tetrathionate was (1) $${\text{S}}_4 {\text{O}}_6^{2 - } + {\text{HS}}^ - \to 2{\text{S}}_2 {\text{O}}_3^{2 - } + {\text{S(O)}} + {\text{H}}^ + $$ added to an anaerobic buffer (pH 8.5) containing excess sulphide. S(O) denotes the zero oxidation state sulphur in the polysulphide mixture S infn sup2- . The addition of formate to the polysulphide solution in the presence of Wolinella succinogenes caused the reduction of polysulphide according to reaction (2). The bacteria grew in a medium containing formate and sulphide, (2) $${\text{HCO}}_2^ - + {\text{S(O)}} + {\text{H}}2{\text{O}} \to {\text{HCO}}_3^ - + {\text{HS}}^ - + {\text{H}}^ + $$ when tetrathionate was continuously added. The cell density increased proportional to reaction (3) which represents the sum of reactions (1) and (3) $${\text{HCO}}_2^ - + {\text{S}}_{\text{4}} {\text{O}}_6^{2 - } + {\text{H}}2{\text{O}} \to {\text{HCO}}_3^ - + 2{\text{S}}_{\text{2}} {\text{O}}_3^{2 - } + 2{\text{H}}^ + $$ (2). The cell yield per mol formate was nearly the same as during growth on formate and elemental sulphur, while the velocity of growth was greater. The specific activities of polysulphide reduction by formate measured with bacteria grown with tetrathionate or with elemental sulphur were consistent with the growth parameters. The results suggest that W. succinogenes grow at the expense of formate oxidation by polysulphide and that polysulphide is an intermediate during growth on formate and elemental sulphur.  相似文献   

13.
The effect of exercise training on heart rate variability (HRV) and improvements in peak oxygen consumption ( peak) was examined in sedentary middle-aged men. The HRV and absolute and relative peak of training (n = 19) and control (n = 15) subjects were assessed before and after a 24-session moderate intensity exercise training programme. Results indicated that with exercise training there was a significantly increased absolute and relative peak (P < 0.005) for the training group (12% and 11% respectively) with no increase for the control group. The training group also displayed a significant reduction in resting heart rate; however, HRV remained unchanged. The trained subjects were further categorized into high (n = 5) and low (n = 5) HRV groups and changes in peak were compared. Improvements in both absolute and relative peak were significantly greater (P > 0.005) in the high HRV group (17% and 20% respectively) compared to the low HRV group (6% and 1% respectively). The groups did not differ in mean age, pretraining oxygen consumption, or resting heart rate. These results would seem to suggest that a short aerobic training programme does not alter HRV in middle-aged men. Individual differences in HRV, however, may be associated with peak response to aerobic training.  相似文献   

14.
The aim of this study was to measure running times to exhaustion (Tlim) on a treadmill at 100% of the minimum velocity which elicits max max in 38 elite male long - distance runners max = 71.4 ± 5.5 ml.kg–1.min–1 and max = 21.8 ± 1.2 km.h–1). The lactate threshold (LT) was defined as a starting point of accelerated lactate accumulation around 4 mM and was expressed in max. Tlim value was negatively correlated with max (r = -0.362, p< 0.05) and max (r = –0.347, p< 0.05) but positively with LT (%v max) (r = 0.378, p < 0.05). These data demonstrate that running time to exhaustion at max in a homogeneous group of elite male long-distance runners was inversely related to max and experimentally illustrates the model of Monod and Scherrer regarding the time limit-velocity relationship adapted from local exercise for running by Hughson et al. (1984) .  相似文献   

15.
Summary A simple heteronuclear relayed E.COSY pulse sequence with a minimum number of pulses is proposed for the quantitative determination of heteronuclear three-bond J-coupling constants in uniformly 13C-enriched polypeptide samples. Numerous heteronuclear three-bond coupling constants, including , , , and , can be determined for each residue from a single heteronuclear relayed E.COSY spectrum. Couplings relevant for stereospecific assignments as well as for the determination of dihedral angles in the amino acid backbone and in side chains are obtained. The method is demonstrated on the uniformly 13C-enriched decapeptide antamanide (-Val1-Pro2-Pro3-Ala4-Phe5-Phe6-Pro7-Pro8-Phe9-Phe10-).  相似文献   

16.
Desulfovibrio vulgaris (strain Marburg) was grown on H2 and sulfate as sole energy source in a chemostat limited by the sulfate supply. The biomass concentration and the sulfate concentration in the culture were determined as a function of the dilution rate. From the data a K S (saturation constant) for sulfate of 10 M, a max of 0.23 h–1, and a of 13 g/mol were calculated. The organism was also grown in chemostat culture on H2 and sulfite, H2 and thiosulfate, and pyruvate (without sulfate). was found to be 35 g/mol, 36 g/mol, and Y pyr max 10 g/mol. The growth yields are discussed with respect to ATP gains in dissimilatory sulfate reduction.  相似文献   

17.
The data processing method of the turbidimetric bioassay of nisin was modified to facilitate its industrial application. The influence of the initial indicator concentration was minimized by a redefined specific dose of the bacteriocin as the quotient between the titer of the added bacteriocin and the initial population density of the indicator in the suspension. It was found that d c = 0.125 μg ml−1 was the critical dose of nisin that can cause a complete inhibition of the indicator, Pediococcus acidilactici UL5, with an initial OD of 0.135. To eliminate the interference of the cell debris, an equation, , exploiting d c, was formulated to obtain the intrinsic survival proportion. The use of the specific dose of the bacteriocin and the intrinsic survival proportion as parameters of the dose/response curve greatly enhanced its repeatability and feasibility. A dual-dosage approach was developed to further simplify the conventional standard dose/response curve method.  相似文献   

18.
Methanobacterium thermoautotrophicum was grown in continuous culture in a fermenter gassed with H2 and CO2 as sole carbon and energy sources, and in a medium which contained either NH4Cl or gaseous N2 as nitrogen source. Growth was possible with N2. Steady states were obtained at various gas flow rates with NH4Cl and with and the maintenance coefficient varied with the gas input and with the nitrogen source. Growth of Methanococcus thermolithotrophicus in continuous culture in a fermenter gassed with H2, CO2 as nitrogen, carbon and energy sources was also examined.Abbreviations molecular growth yield (g dry weight of cells per mol of CH4 evolved) - growth rate (h-1) - D dilution rate (h-1) - rate (h-1); relation of Neijssel and Tempest and of Stouthamer and Bettenhaussen - energy  相似文献   

19.
A mechanistic understanding of perchlorate () entry into plants is important for establishing the human health risk associated with consumption of contaminated produce and for assessing the effectiveness of phytoremediation. To determine whether common soil anions affect uptake and accumulation in higher plants, a series of competition experiments using lettuce (Lactuca sativa L.) were conducted between (50 nM) and (4–12 mM), (1–10 mM), or Cl (5–15 mM) in hydroponic solution. The effects of (0–5 mM) and pH (5.5–7.5) on uptake were also examined. Increasing in solution significantly reduced the amount of taken up by green leaf, butter head, and crisphead lettuces. Sulfate and Cl had no significant effects on uptake in lettuce over the concentrations tested. Increasing pH significantly reduced the amount of taken up by crisphead and green leaf lettuces, whereas increasing significantly reduced uptake in butter head lettuce. The inhibition by across all lettuce genotypes suggests that may share an ion carrier with , and the decrease in uptake with increasing pH or provides macroscopic evidence for cotransport across the plasma membrane.  相似文献   

20.
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