Dynamic cross-linking by alpha-actinin determines the mechanical properties of actin filament networks

We used smooth muscle alpha-actinin to evaluate the contribution of cross-linker dynamics to the mechanical properties of actin filament networks. Recombinant actin-binding domain (residues 2-269) binds actin filaments with a Kd of 1 microM at 25 degrees C, 20 times stronger than actin-binding domain produced by thermolysin digestion of native alpha-actinin (residues 25-257). Between 8 and 25 degrees C the rate constants for recombinant actin-binding domain to bind to (0.8-2.7 microM-1 s-1) and dissociate from (0.2-2.4 s-1) actin filaments depend on temperature. At 8 degrees C actin filaments cross-linked with alpha-actinin are stiff and nearly solid, whereas at 25 degrees C the mechanical properties approach those of actin filaments alone. In these experiments, high actin concentrations kept most of the alpha-actinin bound to actin and temperature varied a single parameter, cross-linker dynamics, because the mechanical properties of pure actin filaments (a viscoelastic gel) or biotinylated actin filaments cross-linked irreversibly by avidin (a stiff viscoelastic solid) depend little on temperature. These results show that the rate of exchange of dynamic cross-links between actin filaments is an important determinant of the mechanical properties of the networks.     

Cooperativity in F-actin: binding of gelsolin at the barbed end affects structure and dynamics of the whole filament

We have studied the effect of gelsolin, a Ca-dependent actin-binding protein, on the microsecond rotational dynamics of actin filaments, using time-resolved phosphorescence (TPA) and absorption anisotropy (TAA) of erythrosin iodoacetamide attached to Cys374 on actin. Polymerization of actin in the presence of gelsolin resulted in substantial increases in the rate and amplitude of anisotropy decay, indicating increased rotational motion. Analysis indicates that the effect of gelsolin cannot be explained by increased rates of overall (rigid-body) rotations of shortened filaments, but reflects changes in intra-filament structure and dynamics. We conclude that gelsolin induces (1) a 10 degrees change in the orientation of the absorption dipole of the probe relative to the actin filament, indicating a conformational change in actin, and (2) a threefold decrease in torsional rigidity of the filament. This result, which is consistent with complementary electron microscopic observations on the same preparations, directly demonstrates long-range cooperativity in F-actin, where a conformational change induced by the binding of a single gelsolin molecule to the barbed end is propagated along inter-monomer bonds throughout the actin filament.     

Loop length, intramolecular diffusion and protein folding

Intramolecular diffusion plays a role in protein folding as shown by kinetic experiments on two alpha-spectrin SH3 domain circular permutants (S19-P20s and N47-D48s), with different poly-glycine loop lengths. Insertion of up to 10 Gly residues does not alter the structure of the folded state nor the overall characteristics of the denatured ensemble. The apparent level of the energy barrier between the denatured and folded species increases linearly with the number of inserted glycines. This suggests that the transition state itself and/or possibly previous transient unstable intermediates are accessed with more difficulty when loop length is increased. The fact that the induced impediment is directly proportional to the number of Gly residues and not to the free energy difference in the folded state indicates that diffusion of different parts of the molecule relative to each other is taking place on going from the denatured ensemble to the transition state. Our results also suggest that transition state ensembles could be more homogenous than recently postulated.     

Nerve length and volume in synaptic vs diffusion neurotransmission: a model

Non-synaptic diffusion neurotransmission (NDN) may be an important factor in brain space and energy conservation, especially within cell assemblies and for mass sustained functions. We have illustrated the extreme cases of total synaptic and total ND neurotransmission for the purpose of noting the differences between the two. For these modeling studies, in which we assume assemblies of 1000 to 100 000 cells supplied by at least one fiber and a single synapse from each of the other cells, each cell assembly would have approximately 200 m to 8000 km of nerve fibers more than when innervated by diffusion. For coeruleo-cortical synaptic innervation, linking each to a common origin (the locus coeruelus), the fiber lengths are 38 cm (1000 cells) to 170 m (100,000 cells). It is likely , however, that neuronal arrays include both 'wireless' (NDN) as well as synaptic intercellular communication systems.     

Tracer diffusion of63Ni in Fe-17 wt pct Cr-12 wt pct Ni

The tracer diffusion of⁶³Ni in Fe-17 Cr-12 Ni by both volume and grain boundary transport has been studied from 600° to 1250°C. The use of an RF sputtering technique for serial sectioning allowed the determination of very small volume diffusion coefficients at the lower temperatures. Volume diffusion of nickel in this alloy was observed to be much slower than in pure iron or austenitic stainless steel at comparable temperatures. The volume diffusion coefficient is described byD _v =8.8 exp (−60,000/RT) cm²/s and grain boundary diffusion is described by σD _gb =3.7×10⁻⁹ exp (−32,000/RT) cm³/s. R. A. PERKINS, formerly Presidential Intern, Metals and Ceramics Division, Oak Ridge National Laboratory, Oak, Ridge, Tenn. 37830, is     

Modeling diffusion through nonuniform concentrations of traps

Cases in which traps are nonuniformly distributed in a solid can be treated by using a modification of the Caskey-Pillinger finite difference method of obtaining solutions to the McNabb-Foster diffusion and trapping equations. The special case of low mobility impurity atom traps that have been previously diffused into one surface of a flat membrane was studied by math modeling with regard to the effect of the traps on the time dependent permeation of faster diffusing atoms. Results obtained for uniformly distributed traps are compared with those expected for nouniformly distributed traps.     

Thomas McKeown and Archibald Cochrane: a journey through the diffusion of their ideas

In the 1970s Thomas McKeown and Archibald L Cochrane were two of the most influential voices in criticizing the dominance of medical thinking. A bibliometric study of the citations to McKeown's The Role of Medicine: Dream, Mirage or Nemesis and Cochrane's Effectiveness and Efficiency: Random Reflections on Health Services was performed from the publication of each book until 1988 to study how their ideas have been disseminated. During the study period 430 papers in the Science Citation Index or the Social Sciences Citation Index cited Cochrane's book, 133 cited McKeown's, and 166 cited both. The citations came mainly from original papers published in journals of internal medicine or public health and epidemiology (35%) and written by authors from the United States or the United Kingdom. Cochrane's book was cited most frequently in medical journals, suggesting a higher degree of penetration of his ideas among medical scientists. Although the dominance of original papers among the citations suggests that these books have been important in stimulating new knowledge, the main problems that McKeown and Cochrane identified--namely, the relatively small impact of clinical medicine on health outcomes and the poor use of scientific methods in clinical practice--are still with us.     

弥散供氧流动特性及其富氧效果

研究了大空间局部圆形出氧口弥散供氧流动特性及其富氧效果.弥散供氧轴向最大速度和氧气轴向最大浓度均随轴向距离增加而衰减,且在轴向x=0-0.6m范围内具有很大速度梯度和浓度梯度.不同出流速度下弥散形成的富氧区域形状是相似的,较小管径下富氧区域下游的浓度轮廓更接近＂半椭圆＂形,弥散范围更大;拟合得到富氧区域外边界扩展半宽度随轴向距离变化的关系式及富氧面积随出流流量变化的关系式.相同流量的富氧采用双出氧口弥散形成的富氧面积比单出氧口弥散形成的富氧面积减少约10%;相同流量的富氧以6mm管径弥散形成的富氧面积比8mm管径的富氧面积增加约10%.