"Adaptive" behavior of ligand-gated ion channels: constraints by thermodynamics

The calcium-induced calcium release channel of the cardiac sarcoplasmic reticulum has been reported to inactivate in a novel manner (termed "adaptation"), which permits reactivation by exposure to successively higher concentrations of calcium. I examined the limitations placed by thermodynamics on the possible kinetic mechanisms for such behavior. The mechanism suggested by Gyorke and Fill, in which the affinity of a calcium-binding site decreases during adaptation, is not thermodynamically feasible for a passive system, but requires an external input of free energy. Possible sources of such energy are 1) metabolic energy, which is excluded by the fact that adaptation was observed in isolated channels in the absence of ATP, or 2) coupling of ion permeation to gating, for which there is currently no evidence. I derived a general limit on the thermodynamic feasibility of a sequence of channel activations and adaptations, irrespective of channel kinetics, from the requirement that the free energy must decrease during the spontaneous evolution of the system from the state existing immediately after a step increase in [Ca2+] to the state of maximum open probability that follows. The opening of the channel must involve an increase in free energy, which must be compensated by the free energy released by the incremental binding of calcium. This requirement leads to a complicated system of inequalities, which was simplified and manipulated algebraically into the form of a linear programming problem. Numerical solution of this problem showed that the sequence of adaptations of the SR channel observed by Gyorke and Fill requires the presence of at least 10 calcium-binding sites on the channel if it is to occur in the absence of exogenous sources of free energy. This indicates either that a large number of calcium-binding sites participate in the regulation of the SR calcium release channel, or that the existing data are significantly flawed with respect to the low open probability in the resting state, the importance of "calcium spike" artifacts from flash photolysis, or both.     

The effect of nonablative laser energy on the ultrastructure of joint capsular collagen

This study was designed to evaluate the effect of laser energy at nonablative levels on the ultrastructure of joint capsular collagen. The femoropatellar joint capsules of six mature New Zealand white rabbits were harvested immediately after death. Specimens were divided into three treatment groups (5, 10, and 15 watts) and one control group. Laser energy was applied using a holmium: YAG laser. Transmission electron microscopy showed significant ultrastructural alterations in collagenous architecture for all laser treatment groups, with increased fibril cross-sectional diameter for each of the treated groups. The fibrils began to lose their distinct edges and their periodical cross-striations at subsequently higher energy densities. A morphometric analysis showed that each subsequently higher laser energy caused a significant increase in collagen fibril diameter. Ultrastructural alteration of collagen fibril architecture caused by the thermal effect of laser energy is probably the dominant mechanism of laser-induced tissue shrinkage.     

Defining the positive tilt test: a study of healthy adults with moderate acute blood loss

STUDY OBJECTIVES: To define a set of orthostatic vital signs that minimize the frequency of false-positives among healthy individuals while maximizing sensitivity in detecting acute moderate blood loss and to determine the sensitivity and specificity of this optimized tilt test in detecting acute moderate blood loss. DESIGN AND INTERVENTION: Postural vital signs were recorded in a standardized manner before and after 450-mL phlebotomy. Paired comparisons were done for a variety of criteria for a positive tilt test using receiver-operating characteristic curves. SETTING AND TYPE OF PARTICIPANTS: Three hundred forty-five healthy euvolemic adult volunteer blood donors were tested at three community blood donation centers over a one-year period. Subjects were prospectively divided into group 1 (less than age 65; 301) and group 2 (age 65 or older; 44). MEASUREMENTS AND MAIN RESULTS: For each combination of pulse and blood pressure in group 1, a change in pulse alone had the same or higher sensitivity with at least the same specificity. Pulse alone was similarly superior in group 2 compared with previously published combinations of pulse and blood pressure. Even the optimized tilt test had limited sensitivity in detecting acute moderate blood loss with high specificity. CONCLUSION: In applying the tilt test to young adults without cardiovascular disease, pulse measurement usually is all that is necessary.     

Sex differences in the relative contributions of nitric oxide and EDHF to agonist-stimulated endothelium-dependent relaxations in the rat isolated mesenteric arterial bed

1. We have used the isolated, buffer-perfused, superior mesenteric arterial bed of male and female rats to assess the relative contributions of nitric oxide (NO) and the endothelium-derived hyperpolarizing factor (EDHF) to endothelium-dependent relaxations to carbachol. 2. Carbachol caused dose-related relaxations of methoxamine-induced tone in mesenteric vascular beds from male rats described by an ED50(M) of 0.43+/-0.15 nmol and a maximum relaxation (Rmax(M) of 89.6+/-1.2% (n=28) which were not significantly different from those observed in mesenteries from female rats (ED50(F)=0.72+/-0.19 nmol and Rax(F)=90.7+/-0.9%; n=22). 3. In the males, the addition of 100 microM NG-nitro-L-arginine methyl ester (L-NAME) caused the dose-response curve to carbachol to be significantly (P<0.001) shifted to the right 15 fold (ED50(M)=6.45+/-3.53 nmol) and significantly (P<0.01) reduced Rmax(M) (79.7+/-2.8%, n=13). By contrast, L-NAME had no effect on vasorelaxation to carbachol in mesenteries from female rats (ED50(f)= 0.89+/-0.19 nmol, Rmax(F)=86.9+/-2.3%, n=9). 4. Raising tone with 60 mM KCl significantly reduced the maximum relaxation to carbachol in mesenteries from male rats 2 fold (Rmax(M)=40.3+/-9.2%, n=4; P<0.001) and female rats by 1.5 fold (Rmax(F)=55.3+/-3.3%, n=6; P<0.001), compared with methoxamine-induced tone. The potency of carbachol was also significantly reduced 1.2 fold in preparations from males (ED50(M)=0.87+/-0.26 nmol; P<0.01) but not the females (ED50(F)=4.04+/-1.46 nmol). In the presence of both 60 mM KCl and L-NAME, the vasorelaxation to carbachol was completely abolished in mesenteries from both groups. 5. The cannabinoid receptor antagonist SR141716A (1 microM), which is also a putative EDHF antagonist, had no significant effect on the responses to carbachol in mesenteries from males or females (ED50(M)=1.41+/-0.74 nmol, Rmax(M)=89.4+/-2.5%, n=7; ED50(F)=2.17+/-0.95 nmol, Rmax(F)=89.9+/-1.8%, n=9). In mesenteries from male rats, in the presence of 100 microM L-NAME, SR141716A significantly (P<0.05) shifted the dose-response curve to carbachol 8 fold further to the right than that seen in the presence of L-NAME alone (ED50(M)= 53.8+/-36.8 nmol) without affecting Rmax(M) (72.4+/-4.8%, n=10). In mesenteries from female rats, the combined presence of L-NAME and SR141716A, significantly (P < 0.01) shifted the dose-response curve to carbachol 7.5 fold, (ED50(F)=6.66+/-2.46 nmol), as compared to L-NAME alone and significantly (P<0.001) decreased Rmax(F) (70.1+/-5.5%, n=8). 6. Vasorelaxations to the nitric oxide donor sodium nitroprusside (SNP), to the endogenous cannabinoid, anandamide (a putative EDHF) and to the ATP-sensitive potassium channel activator, levcromakalim, did not differ significantly between male and female mesenteric vascular beds. 7. The continuous presence of sodium nitroprusside (SNP; 20-60 nM) had no effect on vasorelaxation to carbachol in mesenteries from either males or females. In the presence of L-NAME, SNP significantly (P<0.05) reduced the potency of carbachol 6 fold, without affecting the maximal relaxation in mesenteries from male rats (ED50(M)=40.9+/-19.6 nmol, Rmax(M)=79.4+/-2.5%, n=11). Similarly in mesenteries from female rats, the ED50(F) was also significantly (P<0.01) increased 7 fold (6.24+/-2.02 nmol), while the Rmax(F) was unaffected (81.9+/-11.0%; n=4). 8 The results of the present investigation demonstrate that the relative contributions of agonist-stimulated NO and EDHF to endothelium-dependent relaxations in the rat isolated mesenteric arterial bed, differ between males and females. Specifically, although both NO and EDHF appear to contribute towards endothelium-dependent relaxations in males and females, blockade of NO synthesis alone has no effect in the female. This suggests that EDHF is functionally more important in females; one possible explanation for this is that in the absence of NO, the recently identified ability of EDHF to compensate for the loss of NO, is functio     

Homeless youth and their exposure to and involvement in violence while living on the streets

Sick building syndrome is the term given to a heterogeneous constellation of symptoms that affects workers in modern mechanically ventilated office buildings. Although the cause is unknown, there is evidence that the local environment of the work station is an important determinant of symptoms. In this study, investigators examined the effect of a new, individually controlled ventilation system on workers' symptoms. Investigators studied two groups of workers in one mechanically ventilated office building: (1) a control group at whose worksite no intervention was made and (2) an intervention group. The intervention consisted of installation of a device that allowed each worker control over the ventilation supplied to his or her worksite. Just before, and 4 and 16 mo after installation of this device, workers completed self-administered questionnaires regarding occurrence of symptoms. The new ventilation system resulted in higher air velocities, more variable temperatures, and higher concentrations of airborne dust and fungal spores. Four months after installation, workers with the new ventilation system reported fewer symptoms that were (a) work-related (p < .05) and that were work-related and frequent (p < .05); in addition, they reported fewer symptoms that reduced their capacity to work (p < .01). Sixteen months after installation, workers with the new device reported fewer symptoms than at baseline (although not as significantly), and they indicated that the indoor air quality improved their productivity by 11%, compared with a 4% reduction of productivity among the control group of workers (p < .001). Investigators concluded that the new ventilation system, which provided the workers with individual control over ventilation, was associated with important and sustained reduction in symptoms.     

Preparation of ultrathin reverse osmosis membranes and the attainment of theoretical salt rejection

Cellulose acetate reverse osmosis membranes, 600–2800 A. thick, have been prepared on glass surfaces by dipping a clean glass plate into a dilute solution of cellulose acetate. After drying, the membranes are floated of onto a water surface and placed on molecular filter supports. Theoretical salt rejections, as calculated from the solution-diffusion model of membrane transport for cellulose acetate, were obtained with imperfection-free membranes.     

Colloidal Processing of Zirconium Diboride Ultra‐High Temperature Ceramics

Colloidal processing of the Ultra‐High Temperature Ceramic (UHTC) zirconium diboride (ZrB₂) to develop near?net‐shaping techniques has been investigated. The use of the colloidal processing technique produces higher particle packing that ultimately enables achieving greater densification at lower temperatures and pressures, even pressureless sintering. ZrB₂ suspension formulations have been optimized in terms of rheological behavior. Suspensions were shaped into green bodies (63% relative density) using slip casting. The densification was carried out at 1900°C, 2000°C, and 2100°C, using both hot pressing at 40 MPa and pressureless sintering. The colloidally processed materials were compared with materials prepared by a conventional dry processing route (cold pressed at 50 MPa) and subjected to the same densification procedures. Sintered densities for samples produced by the colloidal route are higher than produced by the dry route (up to 99.5% relative density by hot pressing), even when pressureless sintering is performed (more than 90% relative density). The promising results are considered as a starting point for the fabrication of complex‐shaped components that can be densified at lower sintering temperatures without pressure.     

Conversion of Nuclear Waste to Molten Glass: Cold‐Cap Reactions in Crucible Tests

The feed‐to‐glass conversion, which comprises complex chemical reactions and phase transitions, occurs in the cold cap during nuclear waste vitrification. To investigate the conversion process, we analyzed heat‐treated samples of a simulated high‐level waste feed using X‐ray diffraction, electron probe microanalysis, leaching tests, and residual anion analysis. Feed dehydration, gas evolution, and borate phase formation occurred at temperatures below 700°C before the emerging glass‐forming melt was completely connected. Above 700°C, intermediate aluminosilicate phases and quartz particles gradually dissolved in the continuous borosilicate melt, which expanded with transient foam. Knowledge of the chemistry and physics of feed‐to‐glass conversion will help us control the conversion path by changing the melter feed makeup to maximize the glass production rate.     

Nucleation and crystallization of polypropylene by mineral fillers: relationship to impact strength

The mechanical properties of mineral-filled polypropylene (PP) are determined not only by the size, shape and modulus of the filler particles, but also by microstructure. For example, poor impact strength is correlated with a high capacity for nucleation of crystallization. In the present study, optical microscopy has been used to measure the growth rate of spherulites in PP. In tandem with this, isothermal d.s.c. measurements have been made of the crystallization of the PP filled with talc, calcium carbonate and stearate-coated carbonate at different loading levels. Computer simulation of spherulite growth has been used to derive the number of nucleating sites per unit volume of polymer and, using surface area measurements, the number of sites per unit area of mineral surface was obtained. Values for talc were, as expected, considerably higher than those for carbonate (and especially coated carbonate) fillers. The presence of filler affects not only the nucleation and kinetics of the crystallization process but also the crystallinity and orientation indices and the proportion of β-phase crystallites present. The connection between these factors and impact strength is discussed. It is concluded that impact properties are determined by inter alia crack pinning and blocking by filler particles, stress concentrations at the edges of the filler particles, and the nucleating ability of the filler.