Mechanisms of ion channel activation in primary cultured and clonal cell lines

The effects of hypotonic shock upon membrane C1 permeability of ROS 17/2.8 osteoblast-like cells was investigated using the patch-clamp technique. Hypotonic shock produced cell swelling that was accompanied by large amplitude, outwardly rectifying, currents that were active across the entire physiological range of membrane potentials (-80 to +100 mV). At strong depolarisations (> +50 mV) the currents exhibited time-dependent inactivation that followed a monoexponential time course. The currents were anion selective and exhibited a selectivity sequence of SCN- > I > Br- > Cl- > F- > gluconate. Current activation was unaffected by inhibitors of protein kinase (A (H-89) and tyrosine kinase (tyrphostin A25), and could not be mimicked by elevation of intracellular Ca2+ or activation of protein kinase C. Similarly, disruption of actin filaments by dihydrocytochalsin B, or generation of membrane tension by dipyridamole failed to elicit significant increases in cell chloride permeability. The mechanism of current activation is as yet undetermined. The currents were effectively inhibited by the chloride channel inhibitors NPPB and DIDS but resistant to DPC. A Cl- conductance with similar characteristics was found to be present in mouse primary cultured calvarial osteoblasts. The volume-sensitive Cl- current in ROS 17/2.8 cells was inhibited by arachidonic acid in two distinct phases. A rapid block that developed within 10 s, preceding a slower developing inhibitory phase that occurred approximately 90 s after onset of arachidonate superfusion. Arachidonic acid also induced kinetic modifications of the current which were evident as an acceleration of the time-dependent· inactivation exhibited at depolarised potentials. Inhibitors of cyclo-oxygenases, lipoxygenases and cytochrome P-4S0 were ineffectual against arachidonic acid's effects sugtgesting that arachidonic acid may elicit it's effects directly. Measurements of cell volume under hypotonic conditions showed that ROS 17/2,8 cells could effectively regulate their volume, However, effective inhibitors of the volume-sensitive CI" current drastically impaired this response suggesting that physiologically this current may have a vital role in cell volume regulation, In L6 skeletal myocytes, vasopressin was found to rapidiy hyperpolarise cells. This appears to occur as the result of activation of Ca2+ -sensitive K+ channels in a process dependent upon the presence of extracellular Ca2+.

A preliminary investigation into the impact of a pesticide combination on human neuronal and glial cell lines in vitro

Many pesticides are used increasingly in combinations during crop protection and their stability ensures the presence of such combinations in foodstuffs. The effects of three fungicides, pyrimethanil, cyprodinil and fludioxonil, were investigated together and separately on U251 and SH-SY5Y cells, which can be representative of human CNS glial and neuronal cells respectively. Over 48h, all three agents showed significant reductions in cellular ATP, at concentrations that were more than tenfold lower than those which significantly impaired cellular viability. The effects on energy metabolism were reflected in their marked toxic effects on mitochondrial membrane potential. In addition, evidence of oxidative stress was seen in terms of a fall in cellular thiols coupled with increases in the expression of enzymes associated with reactive species formation, such as GSH peroxidase and superoxide dismutase. The glial cell line showed significant responsiveness to the toxin challenge in terms of changes in antioxidant gene expression, although the neuronal SH-SY5Y line exhibited greater vulnerability to toxicity, which was reflected in significant increases in caspase-3 expression, which is indicative of the initiation of apoptosis. Cyprodinil was the most toxic agent individually, although oxidative stress-related enzyme gene expression increases appeared to demonstrate some degree of synergy in the presence of the combination of agents. This report suggests that the impact of some pesticides, both individually and in combinations, merits further study in terms of their impact on human cellular health.

Development of a neurotoxicity test-system, using human post-mitotic, astrocytic and neuronal cell lines in co-culture

Astrocytes are essential for neuronal function and survival, so both cell types were included in a human neurotoxicity test-system to assess the protective effects of astrocytes on neurons, compared with a culture of neurons alone. The human NT2.D1 cell line was differentiated to form either a co-culture of post-mitotic NT2.N neuronal (TUJ1, NF68 and NSE positive) and NT2.A astrocytic (GFAP positive) cells (∼2:1 NT2.A:NT2.N), or an NT2.N mono-culture. Cultures were exposed to human toxins, for 4 h at sub-cytotoxic concentrations, in order to compare levels of compromised cell function and thus evidence of an astrocytic protective effect. Functional endpoints examined included assays for cellular energy (ATP) and glutathione (GSH) levels, generation of hydrogen peroxide (H2O2) and caspase-3 activation. Generally, the NT2.N/A co-culture was more resistant to toxicity, maintaining superior ATP and GSH levels and sustaining smaller significant increases in H2O2 levels compared with neurons alone. However, the pure neuronal culture showed a significantly lower level of caspase activation. These data suggest that besides their support for neurons through maintenance of ATP and GSH and control of H2O2 levels, following exposure to some substances, astrocytes may promote an apoptotic mode of cell death. Thus, it appears the use of astrocytes in an in vitro predictive neurotoxicity test-system may be more relevant to human CNS structure and function than neuronal cells alone. © 2007 Elsevier Ltd. All rights reserved.

Optimization and characterization of femtosecond laser inscribed in-fiber microchannels for liquid sensing

In-fiber microchannels were fabricated directly in standard single mode fiber using the femtosecond laser inscribe and etch technique. This method of creating in-fiber microchannels offers great versatility since it allows complex three dimensional structures to be inscribed and then etched with hydrofluoric acid. Four in-fiber microchannel designs were experimentally investigated using this technique. Device characteristics were evaluated through monitoring the spectral change while inserting index matching oils into each microchannel - a R.I. sensitivity up to 1.55 dB/RIU was achieved. Furthermore, a simple Fabry-Pérot based refractometer with a R.I. sensitivity of 2.75 nm/RIU was also demonstrated. © 2014 SPIE.

Optimization of gain and SNR in bi-directionally pumped dispersion compensating amplifier modules

In this paper, we present an analysis and optimisation of the performance of bi-directionally pumped dispersion compensation modules acting as simultaneous Raman amplifiers, with optimal configurations for operation with different fibers commercially available. The ratio between forward and backward pump powers for minimum noise influence is obtained in each case, with improvements in the SNR of up to 8 dB when compared to a purely backward-pumped case.

The toxicity of hexanedione isomers in neural and astrocytic cell lines

The metabolite 2,5-hexanedione (HD) is the cause of neurotoxicity linked with chronic n-hexane exposure. Acute exposure to high levels of 2,5-HD, have also shown toxic effects in neuronal cells and non-neuronal cells. Isomers of 2,5-HD, 2,3- and 3,4-HD, added to foodstuffs, are reported to be non-toxic. The acute cytotoxic effects of 2,5-, 2,3- and 3,4-HD were evaluated in neural (NT2.N, SK-N-SH), astrocytic (CCF-STTG1) and non-neural (NT2.D1) cell lines. All the cell lines were highly resistant to 2,5-HD (34-426 mM) at 4-h exposure, although sensitivity was greatest with NT2.D1, then SK-N-SH, NT2.N and finally the CCF-STTG1 line. At 24-h exposure, cell vulnerability increased 5-10-fold. The NT2.D1 cells were again the most sensitive, followed by NT2.N, SK-N-SH and then the CCF-STTG1 cells. 2,3- and 3,4-HD (8-84 mM), were significantly more toxic towards all four cell lines compared with 2,5-HD, after 4-h exposure. After 24-h exposure there was a 12-fold increase in inhibition of MTT turnover in the SK-N-SH cells and a 4-fold increase in the CCF-STTG1 cells, compared with 2,5-HD exposure. 2,3- and 3,4-HD, were significantly less toxic to the NT2.N cells than the SK-N-SH cells after 24-h exposure to the compounds, demonstrating a differing toxin vulnerability between these neural and neuroblastoma cell lines. This study indicates that these non-neuronal and neuronal cells are acutely resistant to 2,5-HD cytotoxicity, whilst the previously unreported sensitivity of all four cell lines to the 2,3- and 3,4- isomers of HD to has been shown to be significantly greater than that of 2,5-HD. © 2006 Elsevier B.V. All rights reserved.

Generalized Scalarizing Problems GENS and GENSLex of Multicriteria Optimization

* This paper is partially supported by the National Science Fund of Bulgarian Ministry of Education and Science under contract № I–1401\2004 "Interactive Algorithms and Software Systems Supporting Multicriteria Decision Making".

Optimization and characterization of femtosecond laser inscribed in-fiber microchannels for liquid sensing

In-fiber microchannels were fabricated directly in standard single mode fiber using the femtosecond laser inscribe and etch technique. This method of creating in-fiber microchannels offers great versatility, since it allows complex three-dimensional structures to be inscribed and then preferentially etched with hydrofluoric acid. In addition, inscription does not require a photosensitive fiber; the modification is induced through nonlinear processes triggered by an ultrashort laser pulse. Four in-fiber microchannel designs were experimentally investigated using this technique - microhole, microslot channel along the core, microslot channel perpendicular to the core and helical channel around the core. Each device design was evaluated through monitoring the optical spectral change while inserting a range of index matching oils into each microchannel; an R.I. sensitivity up to 1.55 dB/RIU was achieved in these initial tests. Furthermore, an all femtosecond laser inscribed Fabry-Pérot-based refractometer with an R.I. sensitivity of 2.75 nm/RIU was also demonstrated. The Fabry-Pérot refractometer was formed by positioning a microchannel between two femtosecond laser inscribed point-by-point fiber Bragg gratings.

Well-Posedness and Conditioning of Optimization Problems

AMS subject classification: 49K40, 90C31.

Power and spectral optimization of random distributed feedback fiber lasers

We present the optimization of power and spectral performances of the random DFB fiber laser using the balance equation set. The numerical results are in good in agreement with experiments. © 2012 OSA.

Multiobjective design optimization of IGBT power modules considering power cycling and thermal cycling

Insulated-gate bipolar transistor (IGBT) power modules find widespread use in numerous power conversion applications where their reliability is of significant concern. Standard IGBT modules are fabricated for general-purpose applications while little has been designed for bespoke applications. However, conventional design of IGBTs can be improved by the multiobjective optimization technique. This paper proposes a novel design method to consider die-attachment solder failures induced by short power cycling and baseplate solder fatigue induced by the thermal cycling which are among major failure mechanisms of IGBTs. Thermal resistance is calculated analytically and the plastic work design is obtained with a high-fidelity finite-element model, which has been validated experimentally. The objective of minimizing the plastic work and constrain functions is formulated by the surrogate model. The nondominated sorting genetic algorithm-II is used to search for the Pareto-optimal solutions and the best design. The result of this combination generates an effective approach to optimize the physical structure of power electronic modules, taking account of historical environmental and operational conditions in the field.

Improved stochastic optimization of railway timetables

We present a general model to find the best allocation of a limited amount of supplements (extra minutes added to a timetable in order to reduce delays) on a set of interfering railway lines. By the best allocation, we mean the solution under which the weighted sum of expected delays is minimal. Our aim is to finely adjust an already existing and well-functioning timetable. We model this inherently stochastic optimization problem by using two-stage recourse models from stochastic programming, building upon earlier research from the literature. We present an improved formulation, allowing for an efficient solution using a standard algorithm for recourse models. We show that our model may be solved using any of the following theoretical frameworks: linear programming, stochastic programming and convex non-linear programming, and present a comparison of these approaches based on a real-life case study. Finally, we introduce stochastic dependency into the model, and present a statistical technique to estimate the model parameters from empirical data.

Optimization of pavement maintenance and rehabilitation programming using shuffled complex evolution algorithm

The major barrier to practical optimization of pavement preservation programming has always been that for formulations where the identity of individual projects is preserved, the solution space grows exponentially with the problem size to an extent where it can become unmanageable by the traditional analytical optimization techniques within reasonable limit. This has been attributed to the problem of combinatorial explosion that is, exponential growth of the number of combinations. The relatively large number of constraints often presents in a real-life pavement preservation programming problems and the trade-off considerations required between preventive maintenance, rehabilitation and reconstruction, present yet another factor that contributes to the solution complexity. In this research study, a new integrated multi-year optimization procedure was developed to solve network level pavement preservation programming problems, through cost-effectiveness based evolutionary programming analysis, using the Shuffled Complex Evolution (SCE) algorithm.^ A case study problem was analyzed to illustrate the robustness and consistency of the SCE technique in solving network level pavement preservation problems. The output from this program is a list of maintenance and rehabilitation treatment (M&R) strategies for each identified segment of the network in each programming year, and the impact on the overall performance of the network, in terms of the performance levels of the recommended optimal M&R strategy. ^ The results show that the SCE is very efficient and consistent in the simultaneous consideration of the trade-off between various pavement preservation strategies, while preserving the identity of the individual network segments. The flexibility of the technique is also demonstrated, in the sense that, by suitably coding the problem parameters, it can be used to solve several forms of pavement management programming problems. It is recommended that for large networks, some sort of decomposition technique should be applied to aggregate sections, which exhibit similar performance characteristics into links, such that whatever M&R alternative is recommended for a link can be applied to all the sections connected to it. In this way the problem size, and hence the solution time, can be greatly reduced to a more manageable solution space. ^ The study concludes that the robust search characteristics of SCE are well suited for solving the combinatorial problems in long-term network level pavement M&R programming and provides a rich area for future research. ^

Eyewitness memory: Balancing the accuracy, precision, and quantity of information through metacognitive monitoring and control

Historically, memory has been evaluated by examining how much is remembered, however a more recent conception of memory focuses on the accuracy of memories. When using this accuracy-oriented conception of memory, unlike with the quantity-oriented approach, memory does not always deteriorate over time. A possible explanation for this seemingly surprising finding lies in the metacognitive processes of monitoring and control. Use of these processes allows people to withhold responses of which they are unsure, or to adjust the precision of responses to a level that is broad enough to be correct. The ability to accurately report memories has implications for investigators who interview witnesses to crimes, and those who evaluate witness testimony. ^ This research examined the amount of information provided, accuracy, and precision of responses provided during immediate and delayed interviews about a videotaped mock crime. The interview format was manipulated such that a single free narrative response was elicited, or a series of either yes/no or cued questions were asked. Instructions provided by the interviewer indicated to the participants that they should either stress being informative, or being accurate. The interviews were then transcribed and scored. ^ Results indicate that accuracy rates remained stable and high after a one week delay. Compared to those interviewed immediately, after a delay participants provided less information and responses that were less precise. Participants in the free narrative condition were the most accurate. Participants in the cued questions condition provided the most precise responses. Participants in the yes/no questions condition were most likely to say “I don’t know”. The results indicate that people are able to monitor their memories and modify their reports to maintain high accuracy. When control over precision was not possible, such as in the yes/no condition, people said “I don’t know” to maintain accuracy. However when withholding responses and adjusting precision were both possible, people utilized both methods. It seems that concerns that memories reported after a long retention interval might be inaccurate are unfounded. ^