Traffic of dynamin within individual Drosophila synaptic boutons relative to compartment-specific markers

Presynaptic terminals contain several specialized compartments, which have been described by electron microscopy. We show in an identified Drosophila neuromuscular synapse that several of these compartments-synaptic vesicle clusters, presynaptic plasma membrane, presynaptic cytosol, and axonal cytoskeleton-labeled by specific reagents may be resolved from one another by laser scanning confocal microscopy. Using a panel of compartment-specific markers and Drosophila shibire(ts1) mutants to trap an intermediate stage in synaptic vesicle recycling, we have examined the localization and redistribution of dynamin within single synaptic varicosities at the larval neuromuscular junction. Our results suggest that dynamin is not a freely diffusible molecule in resting nerve terminals; rather, it appears localized to synaptic sites by association with yet uncharacterized presynaptic components. In shi(ts1) nerve terminals depleted of synaptic vesicles, dynamin is quantitatively redistributed to the plasma membrane. It is not, however, distributed uniformly over presynaptic plasmalemma; instead, fluorescence images show "hot spots" of dynamin on the plasma membrane of vesicle-depleted nerve terminals. We suggest that these dynamin-rich domains may mark the active zones for synaptic vesicle endocytosis first described at the frog neuromuscular junction.     

Refining the level for anticipated hepatotoxicity in acetaminophen poisoning

Treatment of an acetaminophen overdose with N-acetyl cysteine usually is based on the position of the 4-h acetaminophen (APAP) level on the Rumack-Matthew nomogram; however, there is disagreement on the level at which clinically relevant hepatotoxicity occurs. A retrospective review of all acute adult formulation APAP exposures reported to our poison center between 1986 and 1993 was performed and cases corresponding to the "possible risk or toxicity" range on the nomogram were identified. Our current poison center protocol for APAP poisoning does not recommend treatment with N-acetylcysteine (NAC) in low-risk patients if the 4-h serum APAP level or the extrapolated equivalent falls within the possible toxicity range on the nomogram. Seventeen cases met the inclusion criteria for the study and received no NAC; six additional patients met inclusion criteria but received one or two doses of NAC before therapy was discontinued. No patients in either group demonstrated clinical evidence of hepatotoxicity. This pilot study suggests that patients with no risk factors and APAP levels in the "possible risk" range may not require NAC therapy.     

Hamstrings and psoas lengths during normal and crouch gait: implications for muscle-tendon surgery

OBJECTIVE: To study the relative contribution of the lung and the chest wall on the total respiratory system mechanics, gas exchange, and work of breathing in sedated-paralyzed normal subjects and morbidly obese patients, in the postoperative period. SETTING: Policlinico Hospital, University of Milan, Italy. METHODS: In ten normal subjects (normal) and ten morbidly obese patients (obese), we partitioned the total respiratory mechanics (rs) into its lung (L) and chest wall (w) components using the esophageal balloon technique together with airway occlusion technique, during constant flow inflation. We measured, after abdominal surgery, static respiratory system compliance (Cst,rs), lung compliance (Cst,L), chest wall compliance (Cst,w), total lung (Rmax,L) and chest wall (Rmax,w) resistance. Rmax,L includes airway (Rmin,L) and "additional" lung resistance (DR,L). DR,L represents the component due to viscoelastic phenomena of the lung tissue and time constant inequalities (pendelluft). Functional residual capacity (FRC) was measured by helium dilution technique. RESULTS: We found that morbidly obese patients compared with normal subjects are characterized by the following: (1) reduced Cst,rs (p < 0.01), due to lower Cst,L (55.3 +/- 15.3 mL x cm H2O-1 vs 106.6 +/- 31.7 mL x cm H2O-1; p < 0.01) and Cst,w (112.4 +/- 47.4 mL x cm H2O-1 vs 190.7 +/- 45.1 mL x cm H2O-1; p < 0.01); (2) increased Rmin,L (4.7 +/- 3.1 mL x cm H2O x L-1 x s; vs 1.0 +/- 0.8 mL x cm H2O x L-1 x s; p < 0.01) and DR,L (4.9 +/- 2.6 mL x cm H2O x L-1 x s; vs 1.5 +/- 0.8 mL x cm H2O x L-1 x s; p < 0.01); (3) reduced FRC (0.665 +/- 0.191 L vs 1.691 +/- 0.325 L; p < 0.01); (4) increased work performed to inflate both the lung (0.91 +/- 0.25 J/L vs 0.34 +/- 0.08 J/L; p < 0.01) and the chest wall (0.39 +/- 0.13 J/L vs 0.18 +/- 0.04 J/L; p < 0.01); and (5) a reduced pulmonary oxygenation index (PaO2/PAO2 ratio). CONCLUSION: Sedated-paralyzed morbidly obese patients, compared with normal subjects, are characterized by marked derangements in lung and chest wall mechanics and reduced lung volume after abdominal surgery. These alterations may account for impaired arterial oxygenation in the postoperative period.     

<Emphasis Type="Italic">New-Age</Emphasis>: A Negative Bias Temperature Instability-Estimation Framework for Microarchitectural Components

Degradation of device parameters over the lifetime of a system is emerging as a significant threat to system reliability. Among the aging mechanisms, wearout resulting from Negative Bias Temperature Instability (NBTI) is of particular concern in deep submicron technology generations. While there has been significant effort at the device and circuit level to model and characterize the impact of NBTI, the analysis of NBTI’s impact at the architectural level is still at its infancy. To facilitate architectural level aging analysis, a tool capable of evaluating NBTI vulnerabilities early in the design cycle has been developed that evaluates timing degradation due to NBTI. The tool includes workload-based temperature and performance degradation analysis across a variety of technologies and operating conditions, revealing a complex interplay between factors influencing NBTI timing degradation.     

时变滞后Lurie型系统的改进稳定性准则

In this technical note, we present a new stability analysis procedure for ascertaining the delay-dependent stability of a class of Lurie systems with time-varying delay and sector-bounded nonlinearity using Lyapunov-Krasovskii (LK) functional approach. The proposed analysis, owing to the candidate LK functional and tighter bounding of its time-derivative, yields less conservative absolute and robust stability criteria for nominal and uncertain systems respectively. The effectiveness of the proposed criteria over some of the recently reported results is demonstrated using a numerical example.     

Surface Reconstruction based on Lower Dimensional Localized Delaunay Triangulation

We present a fast, memory efficient algorithm that generates a manifold triangular mesh S passing through a set of unorganized points P R ³. Nothing is assumed about the geometry, topology or presence of boundaries in the data set except that P is sampled from a real manifold surface. The speed of our algorithm is derived from a projection-based approach we use to determine the incident faces on a point. We define our sampling criteria to sample the surface and guarantee a topologically correct mesh after surface reconstruction for such a sampled surface. We also present a new algorithm to find the normal at a vertex, when the surface is sampled according our given criteria. We also present results of our surface reconstruction using our algorithm on unorganized point clouds of various models.     

On the pH dependence of protein stability

This paper treats the free energy contribution of ionizable groups to protein stability. A method is presented for the calculation of the pH dependence of the denaturation free energy of a protein, which yields results that can be compared directly to experiment. The first step in the treatment is the determination of the average charges of all the ionizable groups in both the folded and unfolded protein. An expression due to Tanford then relates the pH dependence of the unfolding free energy to the difference in net charge between the two states. In order to determine absolute rather than relative unfolding free energies, it is necessary to calculate the total contribution of ionizable groups to protein stability at some reference pH. This is accomplished through a statistical mechanical treatment similar to the one used previously in the calculation of pKas. The treatment itself is rigorous but it suffers from uncertainties in the pKa calculations. Nevertheless, the overall shape of experimentally observed plots of denaturation free energy as a function of pH are reasonably well reproduced by the calculations. A number of general conclusions that arise from the analysis are: (1) knowledge of titration curves and/or effective pKa values of ionizable groups in proteins is sufficient to calculate the pH dependence of the denaturation free energy with respect to some reference pH value. However, in order to calculate the absolute contribution of ionizable groups to protein stability, it is necessary to also know the intrinsic pKa of each group. This is defined as the pKa of a group in a hypothetical state of the protein where all other groups are neutral. (2) Due to desolvation effects, ionizable groups destabilize proteins, although the effect is strongly dependent on pH. There are however, strongly stabilizing pairwise Coulombic interactions on the surface of proteins. (3) Plots of stability versus pH should not be interpreted in terms of a group whose pKa corresponds to the titration midpoint, but rather to a group with different pKas (that correspond approximately to the titration end points) in each state. (4) Any residual structure in the GuHCl-denatured state of proteins appears to have little effect on the pH dependence of stability. (5) pH-dependent unfolding, for example to the "molten globule" state, appears due to individual groups with anomalous pKas whose locations on the protein surface may determine the nature of the unfolded state.     

Development and use of a two-dimensional interferometer to measure mass flow from a multi-shell Z-pinch gas puff

For gas puff Z-pinches, the K-shell x-ray yield is maximized with the use of a multi-shell nozzle. Optimization of the yield, verification of hydrodynamic models of the nozzle flows, and plausible MHD code modeling of the implosions require data on the radial and axial (R,Z) distribution of mass in the nozzle's flow field. Interferometry is a well-established technique for acquiring such data. We describe the development and use of a two-dimensional interferometer with emphasis on the required data reduction methods. We also show that the instrument can derive the flow from each individual nozzle in a multi-shell system.     

Observations of NBTI-induced atomic-scale defects

A combination of MOSFET gate-controlled diode measurements and a very sensitive electron spin resonance technique called spin-dependent recombination was utilized to observe and identify defect centers generated by a negative bias temperature stress in fully processed SiO/sub 2/-based pMOSFETs. In SiO/sub 2/ devices, the defects include two Si/SiO/sub 2/ interface silicon dangling bond centers (P/sub b0/ and P/sub b1/) and may also include an oxide silicon dangling bond center (E'). The observations indicate that both P/sub b0/ and P/sub b1/ defects play major roles in these SiO/sub 2/-based devices and suggest that E' centers could play an important role.     

Skeletal reduction of boundary value problems

Boundary value problems posed over thin solids are amenable to a dimensional reduction in that one or more spatial variables may be eliminated from the governing equation, resulting in significant computational gains with minimal loss in accuracy. Extant dimensional reduction techniques rely on representing the solid as a hypothetical mid‐surface plus a possibly varying thickness. Such techniques are however hard to automate since the mid‐surface is often ill‐defined and ambiguous. We propose here a skeletal representation based dimensional reduction of boundary value problems. The proposed technique has the computational advantages of mid‐surface reduction, but can be easily automated. A systematic methodology is presented for reducing boundary value problems to lower‐dimensional problems over the skeleton of a solid. The theoretical properties of the proposed method are derived, and supported by representative numerical experiments. Copyright © 2005 John Wiley & Sons, Ltd.