共查询到20条相似文献,搜索用时 15 毫秒
1.
(i) The steady-state kinetic data obtained with purified gizzard and uterus smooth muscle myosins indicated the presence of a plateau region on the substrate-saturation curves. Hill plots of these data provided evidence for mixed positive and negative cooperative interactions. In contrast, when gizzard myosin was prepared according to the method of A. Sobieszek and R.D. Bremel (1975, Eur. J. Biochem.55, 49–60), the saturation curve in the presence of CaATP was hyperbolic and no cooperativity of the binding site(s) was discerned. However, in the presence of MgATP although the curve appeared hyperbolic the Hill plot of the data was biphasic with negative cooperativity at low MgATP concentration, (ii) When thiophosphorylated gizzard myosin was used for kinetic analysis, the plateau region in the presence of MnATP was eliminated from the saturation curve and this curve became hyperbolic. However, in the presence of MgATP, although the plateau was almost eliminated, the saturation curve was still biphasic with either no or greatly reduced negative cooperativity of binding sites at low MgATP concentrations but positive cooperativity of binding at high MgATP concentrations. In addition, the thiophosphorylation of myosin also increased the Km and V of MgATP and MnATP, thus indicating weaker affinity for these substrates with thiophosphorylated myosin. (iii) Gizzard myosin also hydrolyzed other nucleotides (the order of rates being CTP = ITP > ATP = UTP > GTP), therefore saturation kinetics using different nucleotides as substrates was also carried out. The saturation curves with each nucleotide were different i.e., hyperbolic with CTP, sigmoid with GTP, hyperbolic with biphasic Hill plot with ITP, and possessing plateau with UTP. In addition, it was observed that the kinetic pattern with each nucleotide was very sensitive to temperature and pH. 相似文献
2.
Comparison of the myosin and actomyosin ATPase mechanisms of the four types of vertebrate muscles 总被引:16,自引:0,他引:16
The mechanism of ATP hydrolysis by myosin and actomyosin was investigated for the four major classes of vertebrate muscles: fast white (posterior latissimus dorsi), slow red (anterior latissimus dorsi), cardiac and smooth (gizzard). The kinetic behavior of all classes of muscle was consistent with the scheme developed previously for rabbit fast white muscle, but quantitative differences were observed for the rate constants of some of the steps in the hydrolysis cycle. The rate of the hydrolysis step of myosin subfragment-1 was similar for the striated muscles and two to three times smaller for smooth muscle. Two isomerizations of the enzyme occurred in the pathway leading to the formation of the myosin-products intermediate. The rate of dissociation of acto S–1 by ATP was slower for slow muscles and a maximum rate was observed at low temperature. The rate of association of the S-1-products intermediate with actin was equal to the turnover rate of acto S–1 ATPase at low concentrations of actin. The rate of dissociation of ADP from an acto S–1-ADP complex was also much slower for slow muscle. It was shown by Barany (1967) that the maximum turnover rate of actomyosin ATPase (VM) is proportional to the velocity of contraction of the muscle. The only step in the mechanism that is correlated with VM is the apparent second-order rate constant for the formation of a complex of the S-1-product state with actin. The evidence is discussed in terms of a mechanism in which the release of reaction products from actomyosin is the step that is of primary importance in determining the value of VM and the velocity of contraction. 相似文献
3.
Apolinary Sobieszek 《Journal of molecular biology》1982,157(2):275-286
The addition of either smooth muscle or brain tropomyosin to skeletal muscle actoheavy meromyosin (HMM) or acto-myosin subfragment-1 (SF1) produces an activation of the actin-activated ATPase activity up to 100%. This contrasts with the opposite, inhibitory effect produced by skeletal muscle tropomyosin. The degree of activation or inhibition depends on the ionic conditions, which influence the affinities of tropomyosin and HMM or SF1 for actin as well as on the molar ratio of actin to myosin.Enzyme kinetic analysis indicates that the inhibitory effect of skeletal muscle tropomyosin results from an approximately six- to tenfold increase in the apparent affinity (Kapp) of the myosin head for the F-actin-tropomyosin complex with a concomitant six- to tenfold reduction in the maximal turnover rate (Vmax). Thus, there is no direct competition of skeletal muscle tropomyosin and myosin for the same site on actin. Brain tropomyosin has an opposite effect, decreasing the apparent affinity with concomitant increase in the Vmax.The effect of smooth muscle tropomyosin is more complex. At high ratios of myosin to actin this tropomyosin produces the same change in the Kapp as skeletal muscle tropomyosin but yields a value of Vmax that is about twofold higher. At lower molar ratios (below about 1 to 5 myosin subfragments to actin) the activating effect of this tropomyosin remains unchanged while the apparent affinity decreases to that observed for pure F-actin.On the basis of these data as well as from experiments carried out at fixed actin and varying SF1 concentrations, it is concluded that tropomyosins act in general as allosteric un-competitive inhibitors or activators of actomyosin by increasing or reducing the co-operative activation of myosin by actin at the level of product release. 相似文献
4.
Joseph M. Chalovich 《Cell biochemistry and biophysics》1988,12(1):73-85
Smooth muscle contraction is regulated by phosphorylation of myosin and also possibly by the actin associated protein, caldesmon. The properties of caldesmon are discussed and compared with those of tropomyosin-troponin, the well characterized actin-based regulatory system of striated muscle. Caldesmon functions quite differently from tropomyosin-troponin. Under relaxing conditions tropomyosin-troponin does not affect the binding of myosin subfragment-1 to actin. In contrast, caldesmon strongly inhibits the binding of subfragment-1 to actin in the presence of ATP. This inhibition of binding parallels the decrease in ATPase activity that occurs as the caldesmon concentration is increased. Caldesmon has the opposite effect on the two headed myosin subfragment, heavy meromyosin. The apparent binding of skeletal heavy meromyosin increases slightly as the caldesmon concentration is increased, although the rate of ATP hydrolysis is inhibited. It is suggested that in the presence of caldesmon, myosin·ATP does not bind to the productive actin binding site but interacts with a distinct site on actin-caldesmon. This could lead to both an inhibition of ATP hydrolysis and an inrease in resting stiffness of relaxed smooth muscle. 相似文献
5.
L E Greene 《The Journal of biological chemistry》1986,261(3):1279-1285
Binding studies of myosin subfragment one (S-1) to regulated actin in the presence and absence of Ca2+ indicate that, as S-1 binds to regulated actin, tropomyosin-actin units undergo a cooperative transition from a weak to a strong S-1-binding form. Trybus and Taylor (Trybus, K.M., and Taylor, E.W. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 7209-7213) suggested that this transition could be measured by the change in fluorescence of troponin I modified with 4-(N-iodoacetoxyethyl-N-methyl)-7-nitrobenz-2-oxa-1,3-diazole (IANBD). In the present study, this was tested by determining whether the change in fluorescence was proportional to the fraction of tropomyosin-actin units in the strong S-1-binding form as predicted by our model on the cooperative binding of S-1 to regulated actin (Hill, T.L., Eisenberg, E., and Greene, L.E. (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 3186-3190). Experiments were performed both in the presence and absence of Ca2+ by using troponin I modified with either IANBD or 5'-iodoacetamidofluorescein. In the presence of Ca2+, it was found, in agreement with the suggestion of Trybus and Taylor, that the change in fluorescence induced by S-1 was proportional to the fraction of tropomyosin-actin units shifting into the strong S-1 binding form, rather than to the fraction of actin sites having bound S-1. In the absence of Ca2+, the change in fluorescence induced by S-1 also did not reflect the binding of S-1 to regulated actin. However, in contrast to the results in the presence of Ca2+, the change in fluorescence induced by S-1 binding in the absence of Ca2+ was not in agreement with the fraction of tropomyosin-actin units calculated to be in the strong S-1 binding form by the model of Hill et al. Although a more complex model than that of Hill et al. may account for the observed fluorescence changes, it seems equally likely that at least in the absence of Ca2+, the change in fluorescence may be reflecting a more complex behavior than only the transition of tropomyosin-actin units between the weak and strong S-1-binding forms. 相似文献
6.
Three-dimensional reconstruction and averaging of 50 S ribosomal subunits of Escherichia coli from electron micrographs 总被引:3,自引:0,他引:3
The thermal stability and melting kinetics of the α-helical conformation within several regions of the rabbit myosin rod have been investigated. Cyanogen bromide cleavage of long myosin subfragment-2 produced one coiled-coil α-helical fragment corresponding to short subfragment-2 with molecular weight 90,000 (Mr = 45,000) and two fragments from the hinge region with molecular weights of 32,000 to 34,000 (Mr = 16,000 to 17,000) and 24,000 to 26,000 (Mr = 12,000 to 13,000). Optical rotation melting experiments and temperature-jump kinetic studies of long subfragment-2 and its cyanogen bromide fragments show that the hinge and the short subfragment-2 domains melt as quasi-independent co-operative units. The α-helical structure within the hinge has an appreciably lower thermal stability than the flanking short subfragment-2 and light meromyosin regions of the myosin rod. Two relaxation processes for helix-melting, one in the submillisecond range (τf) and the other in the millisecond range (τs), are observed in the light meromyosin and short subfragment-2 regions of the rod, but melting in the hinge domain is dominated by the fast (τf) process. Results suggest that the hinge domain of the subfragment-2 link may be the locus of force generation in a cycling cross-bridge. 相似文献
7.
Calcium-activated tension of skinned muscle fibers of the frog. Dependence on magnesium adenosine triphosphate concentration 总被引:17,自引:6,他引:11
下载免费PDF全文
![点击此处可从《The Journal of general physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
R E Godt 《The Journal of general physiology》1974,63(6):722-739
The influence of MgATP on the Ga++-activated isometric tension of skinned frog muscle fibers was examined in solutions containing: Mg++ = 5 mM, creatine phosphate (CP) = 14.5 mM, creatinephosphokinase (CPK) = 1 mg/ml, total EGTA = 7 mM, CaCl2, KCl, imidazole ≥ 20 mM so that ionic strength = 0.15, pH = 7.00, and MgATP = 2 mM, 0.1 mM, or 20 µM. CP and CPK were necessary for these experiments as determined experimentally by their effect on the tension-Ca++ relation, which was saturated for CP ≥ 14.5 mM. This was interpreted to mean that sufficient CP was present to effectively buffer MgATP intracellularly. Decreasing MgATP shifts the tension-pCa curve to higher pCa (-log Ca++) so that, for half-maximal tension: pCa1/2 = 4.5 for MgATP = 2 mM, pCa1/2 = 5.1 for MgATP = 0.1 mM, and pCa1/2 = 5.8 for MgATP = 20 µM; maximum isometric tension is the same in all cases, however. If MgATP was decreased to 1 µM, tension at Ga++ > 10–8 M was 84% of the maximum Ca-+-activated tension in 2 mM MgATP and increased only slightly to 90% for pCa = 4.5. Weber (1970, In The Physiology and Biochemistry of Muscle as Food, Volume 2, E. J. Briskey, R. G. Cassens, and B. B. Marsh, University of Wisconsin Press, Madison, Wis.), using similar solutions, observed similar shifts in half-maximal calcium activation of rabbit myofibril ATPase rates. In explanation, Weber and Bremel (1971, In Contractility of Muscle Cells and Related Processes, R. J. Podolsky, editor, Prentice-Hall, Inc., Englewood Cliffs, N.J.; Bremel and Weber, 1972, Nat. New Biol., 238:97) have described a mechanism whereby, at low ATP, "rigor complexes" are formed between myosin and thin filament actin and, in turn, alter the calcium affinity of one class of the two Ca++-binding sites on troponin, so that the thin filament is "turned on" for contraction at lower Ca++ levels. Tension data from skinned fibers substantially supports this hypothesis. A stability constant for CaEGTA of 2.62 x 1010 M–1 was determined, with the help of F. N. Briggs, in solutions similar to those used for skinned fibers and was the same for 100 and 300 mM KCl. 相似文献
8.
The flexibility of F-actin complexed with saturating amounts of myosin subfragments has been measured by the use of a dark-field light microscope and a high-sensitivity television camera. When dilute solutions of F-actin complexes were observed in the microscope, single filaments in flexural thermal motion were visible to the eye. Images of the fluctuating filaments were recorded on videotapes using the high-sensitivity camera, and these records were used for the analysis of fluctuation to calculate flexibility in the framework of statistical mechanics of thermal fluctuation in semi-flexible rods. The analysis was carried out by two different methods. In method A, we selected many filaments (the entire length appeared near focus occasionally in the limited period of 10 to 100 seconds), measured the mean square end-to-end distance 〈R2〉 of each filament during the period and also its contour length L, and calculated a parameter λ representing flexibility by the equation given by Landau & Lifshitz (1958): . Then, we obtained a value for λ = 0.040 ± 0.010 μm?1 for the acto-heavy meromyosin filament at 24.0 °C ± 1.0 deg. C, and λ = 0.027 ± 0.005 μm?1 for the acto-tropomyosin-heavy meromyosin filament at the same temperature.In method B, still photographs were taken of the video screen to collect a great number of filaments or parts of filaments which appeared just in focus over their length, and the contour length L of each filament and the angle θ(L) between the tangents at its two ends were measured, on the basis of the assumption that the whole length of each filament was in a plane perpendicular to the direction of view. The data were treated statistically and the results were approximated with 〈cosθ(L)〉 = exp(?λL), which holds for an ensemble of filaments with flexibility λ but in two-dimensional thermal motion (Landau & Lifshitz, 1958). The λ-values obtained by this method for acto-heavy meromyosin and acto-tropomyosin-heavy meromyosin filaments were both in good agreements with those obtained by method A, confirming the reliability of our measurement.F-actin complexed with a saturating amount of myosin subfragment-1 was examined by method B, and its flexibility was shown to be little different from that of acto-heavy meromyosin filaments. 相似文献
9.
Liquid circulation velocity was investigated in three concentric-tube airlift reactors of different scales (RIMP, V L =0.07 m3; RIS-1, V L =2.5 m3; RIS-2, V L =5.20 m3). The effects of top and bottom clearance and resistance in flow pathway at downcomer entrance on the riser liquid superficial velocity, the circulation time, the friction coefficient and flow radial profiles of the gas holdup and the liquid superficial velocity in riser, using water-air as a biphasic system, were studied. It was found that the riser liquid superficial velocity is affected by the analyzed geometrical parameters in different ways, depending on their effects on the pressure loss. The riser liquid superficial velocity, the friction coefficient and the parameters of the drift-flux model were satisfactorily correlated with the bottom spatial ratio (B), gas separation ratio (Y) and downcomer flow resistance ratio (A d /A D ), resulting empirical models, with correlation coefficients greater than 0.85. 相似文献
10.
T I Lin 《Archives of biochemistry and biophysics》1978,185(2):285-299
F-actin has been specifically labeled with a fluorescent probe, dansyl aziridine, at cysteine-373 of the protein. The fluorescence property of the conjugated probe serves as a spectroscopic indicator of several processes in which actin participates. The sulfhydryl modification does not impair the G-F transformation of actin, nor does it affect the complex formation of actin and myosin or the dissociation of the complex by ATP as judged by viscosity measurements. However, both labeled actin and actin modified by N-ethylmaleimide, which also reacts at cysteine-373, stimulate the Mg2+-ATPase of myosin only about 75% as well as unmodified actin. The probe attached to actin exhibits a 65-nm blue shift of its emission maximum from 560 to 495 nm and a sixfold fluorescence enhancement indicating that it is located in a hydrophobic environment. The excitation spectrum of labeled actin indicates that a tryptophan and a tyrosine residue are close to the probe and transfer excitation energy to the dansyl fluorophore. Upon depolymerization of F-actin, the fluorescence intensity of labeled actin increases about 20%. The fluorescence of labeled actin is also enhanced by the addition of EDTA, ATP, and pyrophosphate, but Mg2+ antagonizes this effect reversibly. However, in the presence of 10 mm orthophosphate buffer (pH 7.4) these effects disappear. When labeled F-actin binds with myosin subfragment-1 (SF-1) or heavy meromyosin (HMM), the fluorescence of the actin adduct is enhanced. The fluorescence properties of labeled acto-SF-1 and acto-HMM become insensitive to EDTA and polyphosphates even in the absence of orthophosphate. These results suggest that the two-stranded helical structure of the F-actin filament is stabilized by the presence of phosphate and/or the binding of the myosin “head”. 相似文献
11.
Hanneke Borgdorff Stuart D. Armstrong Hanne L. P. Tytgat Dong Xia Gilles F. Ndayisaba Jonathan M. Wastling Janneke H. H. M. van de Wijgert 《PloS one》2016,11(3)
Background
A Lactobacillus-dominated cervicovaginal microbiota (VMB) protects women from adverse reproductive health outcomes, but the role of L. iners in the VMB is poorly understood. Our aim was to explore the association between the cervicovaginal L. iners and L. crispatus proteomes and VMB composition.Methods
The vaginal proteomes of 50 Rwandan women at high HIV risk, grouped into four VMB groups (based on 16S rDNA microarray results), were investigated by mass spectrometry using cervicovaginal lavage (CVL) samples. Only samples with positive 16S results for L. iners and/or L. crispatus within each group were included in subsequent comparative protein analyses: Lactobacillus crispatus-dominated VMB cluster (with 16S-proven L. iners (ni) = 0, and with 16S-proven L. crispatus (nc) = 5), L. iners-dominated VMB cluster (ni = 11, nc = 4), moderate dysbiosis (ni = 12, nc = 2); and severe dysbiosis (ni = 8, nc = 2). The relative abundances of proteins that were considered specific for L. iners and L. crispatus were compared among VMB groups.Results
Forty Lactobacillus proteins were identified of which 7 were specific for L. iners and 11 for L. crispatus. The relative abundances of L. iners DNA starvation/stationary phase protection protein (DPS), and the glycolysis enzymes glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and glucose-6-phosphate isomerase (GPI), were significantly decreased in women with L. iners-containing dysbiosis compared to women with a L. iners-dominated VMB, independent of vaginal pH and L. iners abundance. Furthermore, L. iners DPS, GAPDH, GPI, and fructose-bisphosphate aldolase (ALDO) were significantly negatively associated with vaginal pH. Glycolysis enzymes of L. crispatus showed a similar negative, but nonsignificant, trend related to dysbiosis.Conclusions
Most identified Lactobacillus proteins had conserved intracellular functions, but their high abundance in CVL supernatant might imply an additional extracellular (moonlighting) role. Our findings suggest that these proteins can be important in maintaining a Lactobacillus-dominated VMB. Functional studies are needed to investigate their roles in vaginal bacterial communities and whether they can be used to prevent vaginal dysbiosis. 相似文献12.
Cooperativity of actin-activated ATPase of gizzard heavy meromyosin in the presence of gizzard tropomyosin 总被引:2,自引:0,他引:2
The mechanism for the potentiation of the actin-activated ATPase of smooth muscle myosin by tropomyosin is investigated using smooth muscle actin, tropomyosin, and heavy meromyosin. In the presence of tropomyosin, an increase in Vmax occurs with no effect on KATPase and Kbinding at 20 mM ionic strength. Utilizing N-ethylmaleimide-treated subfragment-1, which forms rigor complexes with actin in the presence of ATP but does not have ATPase activity, experiments were carried out to determine if the tropomyosin-actin complex exists in both the turned-off and turned-on forms as in the skeletal muscle system. At both 60 and 100 mM ionic strengths, the presence of rigor complexes on the smooth muscle actin filament containing bound tropomyosin causes a 2-3-fold increase in Vmax and about a 3-fold increase in KATPase, resulting in about a 4-fold increase in ATPase activity at moderate actin concentration. The increase in KATPase is correlated with an increase in Kbinding. The finding that rigor complexes increase Vmax and the binding constant for heavy meromyosin to tropomyosin-actin at an ionic strength close to physiological conditions indicates that the tropomyosin-actin complex can be turned on by rigor complexes in a cooperative manner. However, in contrast to the situation in the skeletal muscle system, the increase in KATPase is associated with a corresponding increase in Kbinding. Furthermore, there is only a 3-fold increase in KATPase in the smooth muscle system rather than a 10-fold increase as in the skeletal muscle system. 相似文献
13.
Δ53β hydroxysteroid dehydrogenase activity transforms biologically inactive Δ53β hydroxy steroids into the active Δ43-keto products (e.g. pregnenolone to progesterone). Using a cytochemical procedure which allows for the continuous microdensitometric monitoring of an enzyme reaction as it proceeds and a well described cytochemical assay for Δ53β HSD we have analysed the initial velocity rates (Vo) for dehydroepiandrosterone (DHEA) binding to this enzyme in regressing (i.e. 20α hydroxy steroid dehydrogenase positive) corpus luteum (CL) cells in unfixed tissue sections (5 μm) of the dioestrous and proestrous rat ovary. The results are mean ± S.E.M. The relationship between DHEA concentration (0 to 50 μM) and Δ53β HSD activity in the dioestrous corpora lutea was sigmoidal and had an atypical 1/Vo versus 1/S plot, the x intercept being positive. Using a 1/Vo versus 1/S2 plot the Vmax was determined to be 1·0 ± 0·08 μmol min?1 mg?1 CL (n = 6). The Hill constant was 2·7 ± 0·02 (n = 6) suggesting a high degree of positive co-operativity for DHEA binding. The S concentration for half maximal activity was 17 ± 1 μmoles (n = 6). In the corpora lutea cells of the proestrous ovary, the Vmax for DHEA transformation was unchanged (0·95 ± 0·04 μmol min?1 mg?1, n = 3) whilst the S0·5 was significantly increased to 27 ± 0·1 (p < 0·01, n = 3). The Hill constant remained positive being 2·9 ± 0·2 (n = 3). NAD+ binding to 3β HSD in regressing corpora lutea of the proestrous ovary has been demonstrated previously to be hyperbolic and fit the classical Michaelis-Menten model.1 Extending the analysis of NAD+ binding to the regressing corpus luteum of the dioestrous rat ovary revealed similar kinetic characteristics to that seen with the proestrous enzyme, the apparent Vmax and Km being 0·84 ± 0·04 μmol min?1 mg?1 CL (n = 3) and 27 ± 7 μmol 1?1 (n = 3) respectively. The Hill constant was 1·1 ± 0·03 (n = 3), indicating no co-operativity of co-factor binding. 相似文献
14.
Lilian Motta Cantanhêde Cipriano Ferreira da Silva Júnior Marcos Massayuki Ito Kátia Paula Felipin Roberto Nicolete Juan Miguel Villalobos Salcedo Renato Porrozzi Elisa Cupolillo Ricardo de Godoi Mattos Ferreira 《PLoS neglected tropical diseases》2015,9(9)
Tegumentary Leishmaniasis (TL) is endemic in Latin America, and Brazil contributes approximately 20 thousand cases per year. The pathogenesis of TL, however, is still not fully understood. Clinical manifestations vary from cutaneous leishmaniasis (CL) to more severe outcomes, such as disseminated leishmaniasis (DL), mucosal leishmaniasis (ML) and diffuse cutaneous leishmaniasis (DCL). Many factors have been associated with the severity of the disease and the development of lesions. Recent studies have reported that the presence of Leishmania RNA virus 1 infecting Leishmania (Leishmania RNA virus 1, LRV1) is an important factor associated with the severity of ML in experimental animal models. In the present study, 156 patients who attended Rondonia''s Hospital of Tropical Medicine with both leishmaniasis clinical diagnoses (109 CL; 38 ML; 5 CL+ML; 3 DL and 1 DCL) and molecular diagnoses were investigated. The clinical diagnosis were confirmed by PCR by targeting hsp70 and kDNA DNA sequences and the species causing the infection were determined by HSP70 PCR-RFPL. The presence of LVR1 was tested by RT-PCR. Five Leishmania species were detected: 121 (77.6%) samples were positive for Leishmania (Viannia) braziliensis, 18 (11.5%) were positive for Leishmania (V.) guyanensis, 3 (1.8%) for Leishmania (V.) lainsoni, 2 (1.3%) for Leishmania (Leishmania) amazonensis and 2 (1.3%) for Leishmania (V.) shawi. Six (3.9%) samples were positive for Leishmania sp. but the species could not be determined, and 4 (2.6%) samples were suggestive of mixed infection by L. (V.) braziliensis and L. (V.) guyanensis. The virus was detected in L. braziliensis (N = 54), L. guyanensis (N = 5), L. amazonensis (N = 2), L. lainsoni (N = 1) and inconclusive samples (N = 6). Patients presenting with CL+ML, DL and DCL were excluded from further analysis. Association between the presence of the virus and the disease outcome were tested among the remaining 147 patients (CL = 109 and ML = 38). Of them, 71.1% (n = 27) mucosal lesions were positive for LRV1, and 28.9% (n = 11) were negative. In cutaneous lesions, 36.7% (n = 40) were positive and 63.3% (n = 69) were negative for LRV1. The ratio P(ML|LRV1+)/P(ML|LRV1-) was 2.93 (CI95% 1.57…5.46; p<0.001), thus corroborating the hypothesis of the association between LRV1 and the occurrence of mucosal leishmaniasis, as previously described in animal models; it also indicates that LRV1 is not the only factor contributing to the disease outcome. 相似文献
15.
We describe the first dynamic programming algorithm that computes the expected degree for the network, or graph G = (V, E) of all secondary structures of a given RNA sequence a = a
1, …, a
n. Here, the nodes V correspond to all secondary structures of a, while an edge exists between nodes s, t if the secondary structure t can be obtained from s by adding, removing or shifting a base pair. Since secondary structure kinetics programs implement the Gillespie algorithm, which simulates a random walk on the network of secondary structures, the expected network degree may provide a better understanding of kinetics of RNA folding when allowing defect diffusion, helix zippering, and related conformation transformations. We determine the correlation between expected network degree, contact order, conformational entropy, and expected number of native contacts for a benchmarking dataset of RNAs. Source code is available at http://bioinformatics.bc.edu/clotelab/RNAexpNumNbors. 相似文献
16.
Theoretical models for cooperative steady-state ATPase activity of myosin subfragment-1 on regulated actin 总被引:3,自引:0,他引:3
下载免费PDF全文
![点击此处可从《Biophysical journal》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Recent theoretical work on the cooperative equilibrium binding of myosin subfragment-1-ADP to regulated actin, as influenced by Ca2+, is extended here to the cooperative steady-state ATPase activity of myosin subfragment-1 on regulated actin. Exact solution of the general steady-state problem will require Monte Carlo calculations. Three interrelated special cases are discussed in some detail and sample computer (not Monte Carlo) solutions are given. The eventual objective is to apply these considerations to in vitro experimental data and to in vivo muscle models. 相似文献
17.
Background
Severe impairment of the major respiratory muscles resulting from tetraplegia reduces respiratory function, causing many people with tetraplegia to require mechanical ventilation during the acute stage of injury. Abdominal Functional Electrical Stimulation (AFES) can improve respiratory function in non-ventilated patients with sub-acute and chronic tetraplegia. The aim of this study was to investigate the clinical feasibility of using an AFES training program to improve respiratory function and assist ventilator weaning in acute tetraplegia.Methods
AFES was applied for between 20 and 40 minutes per day, five times per week on four alternate weeks, with 10 acute ventilator dependent tetraplegic participants. Each participant was matched retrospectively with a ventilator dependent tetraplegic control, based on injury level, age and sex. Tidal Volume (VT) and Vital Capacity (VC) were measured weekly, with weaning progress compared to the controls.Results
Compliance to training sessions was 96.7%. Stimulated VT was significantly greater than unstimulated VT. VT and VC increased throughout the study, with mean VC increasing significantly (VT: 6.2 mL/kg to 7.8 mL/kg VC: 12.6 mL/kg to 18.7 mL/kg). Intervention participants weaned from mechanical ventilation on average 11 (sd: ± 23) days faster than their matched controls.Conclusion
The results of this study indicate that AFES is a clinically feasible technique for acute ventilator dependent tetraplegic patients and that this intervention may improve respiratory function and enable faster weaning from mechanical ventilation.Trial Registration
ClinicalTrials.gov NCT02200393 相似文献18.
Jingyi Ren Jing Zhang Ning Xu Guanping Han Qiang Geng Junxian Song Sufang Li Jianqing Zhao Hong Chen 《PloS one》2013,8(12)
Aims
MicroRNAs (miRNAs) play important roles in the pathogenesis of cardiovascular diseases. Circulating miRNAs were recently identified as biomarkers for various physiological and pathological conditions. In this study, we aimed to identify the circulating miRNA fingerprint of vulnerable coronary artery disease (CAD) and explore its potential as a novel biomarker for this disease.Methods and Results
The Taqman low-density miRNA array and coexpression network analyses were used to identify distinct miRNA expression profiles in the plasma of patients with typical unstable angina (UA) and angiographically documented CAD (UA group, n = 13) compared to individuals with non-cardiac chest pain (control group, n = 13). Significantly elevated expression levels of miR-106b/25 cluster, miR-17/92a cluster, miR-21/590-5p family, miR-126*, and miR-451 were observed in UA patients compared to controls. These findings were validated by real-time PCR in another 45 UA patients, 31 stable angina patients, and 37 controls. In addition, miR-106b, miR-25, miR-92a, miR-21, miR-590-5p, miR-126* and miR-451 were upregulated in microparticles (MPs) isolated from the plasma of UA patients (n = 5) compared to controls (n = 5). Using flow cytometry and immunolabeling, we further found that Annexin V+ MPs were increased in the plasma samples of UA patients compared to controls, and the majority of the increased MPs in plasma were shown to be Annexin V+ CD31+ MPs. The findings suggest that Annexin V+ CD31+ MPs may contribute to the elevated expression of the selected miRNAs in the circulation of patients with vulnerable CAD.Conclusion
The circulating miRNA signature, consisting of the miR-106b/25 cluster, miR-17/92a cluster, miR-21/590-5p family, miR-126* and miR-451, may be used as a novel biomarker for vulnerable CAD.Trial Registration
Chinese Clinical Trial Register, ChiCTR-OCH-12002349. 相似文献19.
Lennart Hilbert Shivaram Cumarasamy Nedjma B. Zitouni Michael C. Mackey Anne-Marie Lauzon 《Biophysical journal》2013
Naturally occurring groups of muscle myosin behave differently from individual myosins or small groups commonly assayed in vitro. Here, we investigate the emergence of myosin group behavior with increasing myosin group size. Assuming the number of myosin binding sites (N) is proportional to actin length (L) (N = L/35.5 nm), we resolve in vitro motility of actin propelled by skeletal muscle myosin for L = 0.2–3 μm. Three distinct regimes were found: L < 0.3 μm, sliding arrest; 0.3 μm ≤ L ≤ 1 μm, alternation between arrest and continuous sliding; L > 1 μm, continuous sliding. We theoretically investigated the myosin group kinetics with mechanical coupling via actin. We find rapid actin sliding steps driven by power-stroke cascades supported by postpower-stroke myosins, and phases without actin sliding caused by prepower-stroke myosin buildup. The three regimes are explained: N = 8, rare cascades; N = 15, cascade bursts; N = 35, continuous cascading. Two saddle-node bifurcations occur for increasing N (mono → bi → mono-stability), with steady states corresponding to arrest and continuous cascading. The experimentally measured dependence of actin sliding statistics on L and myosin concentration is correctly predicted. 相似文献
20.
The fluorescence lifetime of 1,N6-ethenoadenosine diphosphate (?-ADP) is 33 ns when bound to F-actin at 4 °C. When heavy meromyosin or myosin subfragment-1 binds to the F-actin filament, the lifetime of ?-ADP drops, reaching 29 ns when every actin monomer is bound to a myosin head. The change in lifetime is a consequence of cooperative conformational changes among the actin monomers. The results of these experiments support the contention that there are differences in the ways in which the two heads of the myosin molecule interact with the actin filament. 相似文献