U18666A-mediated apoptosis in cultured murine cortical neurons : role of caspases, calpains and kinases

Studies have suggested that cholesterol imbalance in the brain might be related to the development of neurological disorders such as Alzheimer's disease and Niemann–Pick disease type C. Previously, we have reported that U18666A, a cholesterol transport-inhibiting agent, leads to apoptosis and intracellular cholesterol accumulation in primary cortical neurons. In this study, we examined the effects of U18666A-mediated neuronal apoptosis, and found that chronic exposure to U18666A led to the activation of caspases and calpains and hyperphosphorylation of tau. Tau hyperphosphorylation is regulated by several kinases that phosphorylate specific sites of tau in vitro. Surprisingly, the kinase activity of cyclin-dependent kinase 5 decreased in U18666A-treated cortical neurons whereas its protein level remained unchanged. The amount of glycogen synthase kinase 3 and mitogen-activated protein kinases were found to decrease in their phosphorylated states by Western blot analysis. Gene transcription was further studied using microarray analysis. Genes encoding for kinases and phosphatases were differentially expressed with most up-regulated and some down-regulated in expression upon U18666A treatment. The activation of cysteine proteases and cholesterol accumulation with tauopathies may provide clues to the cellular mechanism of the inhibition of cholesterol transport-mediated cell death in neurodegenerative diseases.

Copper/zinc superoxide dismutase is phosphorylated and modulated specifically by granulocyte-colony stimulating factor in myeloid cells.

Using two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2-D SDS-PAGE) of 32P-labeled cytosolic and membrane extracts, we identified a 21.5 kDa phosphoprotein with an isoelectric point of 6.0 in NFS-60 cells that was phosphorylated maximally at 15 min by treatment with granulocyte-colony stimulating factor (G-CSF) but not with interlevkin-3 (IL-3) or colony-stimulating factor-1 (macrophage-colony stimulating factor (CSF-1 (M-CSF)). The phosphorylation of this protein, designated 21.5/6.0, was unaffected by a series of antiproliferative agents [32]. These findings suggested that the 21.5/6.0 phosphoprotein may be involved in specific G-CSF-mediated biological responses such as activation and/or differentiation. We sought to characterize this 21.5/6.0 by a novel combination of 2-D SDS-PAGE and hydroxyapatite (HTP)-chromatography. Amino acid sequence determination of 21.5/6.0 revealed it to share a high level of homology with copper/zinc superoxide dismutase (Cu/Zn-SOD), indicating that a Cu/Zn-SOD is phosphorylated following treatment with G-CSF. This is the first report of the phosphorylation and possible involvement of Cu/Zn-SOD protein in granulocyte activation/differentiation events. In addition, Cu/Zn-SOD levels and activity were diminished by G-CSF but not IL-3 treatment. This new protocol combining 2-D SDS-PAGE and HTP-chromatography allows the characterization of low abundance phosphoproteins involved in the cellular responses to G-CSF and presumably to other cytokines/growth factors.

Time-to-Collision (TTC) judgements with offsize objects show that Tau needs to be intergrated with familiar size to explain TTC performance

Observers judged TTC with computer-generated displays simulating an approaching object in three familiar-size conditions:

(i) Real-size (smaller, larger objects depicted as tennis, soccer balls respectively).
(ii) Off-size (smaller, larger objects depicted as soccer, tennis balls respectively).
(iii) Ambiguous-size (smaller, larger objects depicted as texture-less black balls of different size).

Displays simulated objects approaching observersí viewpoint from 24.96 m, and disappearing at 5.76 m. Manipulation of approach velocities (4.8-19.2 msec-1) produced viewing times from 1.0 to 4.0 sec, and delays between object disappearance and tau-based TTC ranging from 0.3 to 1.2 sec. Motion characteristics of smaller and larger objects in the three familiar-size conditions simulated those of approaching real-sized tennis and soccer balls respectively; that is, for each approach velocity, tau‚-based TTC was the same across the three conditions for smaller and larger objects.

Results showed that, consistent with the proposition of tau-determined TTC, TTC estimates in the real-size condition were uninfluenced by object size. This is contrary to previous reports that TTC for larger objects is underestimated relative to TTC for smaller objects. However, such size-dependent TTC differences were found in the ambiguous-size condition, with even larger differences in the off-size condition; TTCs for the ëlargerí tennis ball were much less than TTCs to the ësmallerí soccer ball compared to corresponding TTCs in the ambiguous-size condition. These results are problematic for the proposition that tau solely determines TTC. We discuss the role of perceptual learning in resolving this problem.

Improving the kinematic performance of the SCARA-Tau PKM

One well acknowledged drawback of traditional parallel kinematic machines (PKMs) is that the ratio of accessible workspace to robot footprint is small for these structures. This is most likely a contributing reason why relatively few PKMs are used in industry today. The SCARA-Tau structure is a parallel robot concept designed with the explicit goal of overcoming this limitation and developing a PKM with a workspace similar to that of a serial type robot of the same size. This paper shows for the first time how a proposed variant of the SCARA-Tau PKM can improve the usability of this robot concept further by significantly reducing the dependence between tool platform position and orientation of the original concept. The inverse kinematics of the proposed variant is derived and a comparison is made between this structure and the original SCARA-Tau concept, both with respect to platform orientation changes and workspace.

A comparison of the yaw constraining performance of SCARA-tau parallel manipulator variants via screw theory

The SCARA-Tau parallel manipulator was derived with the objective to overcome the limited workspace-to-footprint ratio of the DELTA parallel manipulator while maintaining its many benefits. The SCARA-Tau family has later been extended and a large number of variants have been proposed. In this paper, we analyse four of these variants, which together encompass the main differences between all the proposed SCARA-Tau manipulators. The analysed manipulator variants utilise an identical arrangement of five of the six linkages connecting the actuated arms and the manipulated platform and exhibit the same input-output Jacobian. The normalised reciprocal product between the wrench of the sixth linkage and the twist of the platform occurring without this linkage provides a measure on how effectively the sixth linkage constrains the manipulated platform. A comparison of the manipulator variants with respect to this measure demonstrates each variants suitability for specific applications.

Effects of approach velocity and foot-target characteristics on the visual regulation of step length

Two questions emerge from the literature concerning the perceptual-motor processes underlying the visual regulation of step length. The first concerns the effects of velocity on the onset of visual control (VCO), when visual regulation of step length begins during goal-directed locomotion. The second concerns the effects of different obstacles such as a target or raised surface on step length regulation. In two separate experiments, participants (Experiment 1 & 2: n=12, 6 female, 6 male) walked, jogged, or sprinted towards an obstacle along a 10 m walkway, consisting of two marker-strips with alternating black and white 0.50 m markings. Each experiment consisted of three targeting or obstacle tasks with the requirement to both negotiate and continue moving (run-through) through the target. Five trials were conducted for each task and approach speed, with trials block randomised between the six participants of each gender. One 50 Hz video camera panned and filmed each trial from an elevated position, adjacent to the walkway. Video footage was digitized to deduce the gait characteristics. Results for the targeting tasks indicate a linear relationship between approach velocity and accuracy of final foot placement (r=0.89). When foot placement was highly constrained by the obstacle step length shortened during the entire approach. VCO was found to occur at an earlier tau-margin for lower approach velocities for both experiments, indicating that the optical variable ‘tau' is affected by approach velocity. A three-phase kinematic profile was found for all tasks, except for the take-off board condition when sprinting. Further research is needed to determine whether this velocity affect on VCO is due to ‘whole-body' approach velocity or whether it is a function of the differences between gait modes.

The effects of target length on the visual control of step length for hard and soft impacts

Adjustments to gait were examined when positioning the foot within a narrow target at the end of an approach for two impact conditions, hard and soft. Participants (6 M, 6 F) ran toward a target of three lengths along a 10-m walkway consisting of two marker strips with alternating black and white 0.5-m markings. Five trials were conducted for each target length and impact task, with trials block randomized between the 6 participants of each gender. A 50-Hz digital video camera panned and filmed each trial from an elevated position adjacent to the walkway. Video footage was digitized to deduce the gait characteristics. A linear speed/accuracy tradeoff between target length and approach time was found for both impact tasks (hard, r = 0.99, p < 0.01; soft, r = 0.96, p < 0.05). For the hard-impact task, visual control time increased linearly (r = 0.99, p < 0.05) when whole-body approach velocity decreased. Visual control time was unaffected by whole-body approach velocity in the soft-impact task. A constant tau-margin of 1.08 describes the onset of visual control when approaching a target while running, with the control of braking during visual control described by a tau-dot of –0.85. Further research is needed to examine the control of braking in different targeting tasks.

The salt wedge position in a bar-blocked estuary subject to pulsed inflows

A series of laboratory experiments were carried out to investigate the response of a bar-blocked, saltwedge estuary to the imposition of both steady freshwater inflows and transient inflows that simulate storm events in the catchment area or the regular water releases from upstream reservoirs. The trapped salt water forms a wedge within the estuary, which migrates downstream under the influence of the freshwater inflow. The experiments show that the wedge migration occurs in two stages, namely (i) an initial phase characterized by intense shear-induced mixing at the nose of the wedge, followed by (ii) a relatively quiescent phase with significantly reduced mixing in which the wedge migrates more slowly downstream.

Provided that the transition time t_T between these two regimes satisfies t_T>g′h⁴L/q³α, as was the case for all our experiments and is likely to be the case for most estuaries, then the transition occurs at time t_T=1.2(gα³L⁶/g′³q²)^1/6, where g′=gΔρ/ρ0 is the reduced gravity, g the acceleration due to gravity, Δρ the density excess of the saline water over the density ρ₀ of the freshwater, q the river inflow rate per unit width, and L and α are the length and bottom slope of the estuary, respectively.

A simple model, based on conversion of the kinetic energy of the freshwater inflow into potential energy to mix the salt layer, was developed to predict the displacement xw over time t of the saltwedge nose from its initial position. For continuous inflows subject to t<t_T, the model predicts the saltwedge displacement as x_w/h=1.1 (t/&tau;)^1/3, where the normalizing length and time scales are h=(q²/g)^1/3 and &tau;=g′α²h4L/q³, respectively. For continuous inflows subject to t>tT, the model predicts the displacement as xw/h=0.45N1/6(t/&tau;)1/6/α, where N=q²/g′h²L is a non-dimensional number for the problem. This model shows very good agreement with the experiments. For repeated, pulsed discharges subject to t<t_T, the saltwedge displacement is given by (xw/h)3−(x0/h)(xw/h)2=1.3t/&tau;, where x₀ is the initial displacement following one discharge event but prior to the next event. For pulsed discharges subject to t>t_T, the displacement is given by (xw/h)⁶−(x0/h)(xw/h)⁵=0.008N(t/&tau;)/α⁶. This model shows very good agreement with the experiments for the initial discharge event but does systematically underestimate the wedge position for the subsequent pulses. However, the positional error is less than 15%.

Blind source separation based on phase and frequency redundancy of cyclostationary signals

This paper presents a new method for blind source separation by exploiting phase and frequency redundancy of cyclostationary signals in a complementary way. It requires a weaker separation condition than those methods which only exploit the phase diversity or the frequency diversity of the source signals. The separation criterion is to diagonalize a polynomial matrix whose coefficient matrices consist of the correlation and cyclic correlation matrices, at time delay .TAU. = 0, of multiple measurements. An algorithm is proposed to perform the blind source separation. Computer simulation results illustrate the performance of the new algorithm in comparison with the existing ones.