Optimization of immunocytochemistry in cytology: comparison of two protocols for fixation and preservation on cytospin and smear preparations

Objective: A new protocol for fixation and slide preservation was evaluated in order to improve the quality of immunocytochemical reactions on cytology slides. Methods: The quality of immunoreactions was evaluated retrospectively on 186 cytology slides (130 direct smears, 56 cytospins) prepared from different cytology samples. Ninety-three of the slides were air dried, stored at -20 °C and fixed in acetone for 10 minutes (Protocol 1), whereas the other 93 were immediately fixed in methanol at -20 °C for at least 30 minutes, subsequently protected with polyethylene glycol (PEG) and stored at room temperature (Protocol 2). Immunocytochemical staining, with eight primary antibodies, was performed on a Ventana BenchMark Ultra instrument using an UltraView Universal DAB Detection Kit. The following parameters were evaluated for each immunoreaction: morphology preservation, intensity of specific staining, background and counterstain. The slides were blinded and independently scored by four observers with marks from 0 to 20. Results: The quality of immunoreactions was better on methanol-fixed slides protected with PEG than on air-dried slides stored in the freezer: X¯ = 14.44 ± 3.58 versus X¯ = 11.02 ± 3.86, respectively (P < 0.001). Conclusion: Immediate fixation of cytology slides in cold methanol with subsequent application of PEG is an easy and straightforward procedure that improves the quality of immunocytochemical reactions and allows the storage of the slides at room temperature.

Economic, environmental and mixed objective functions in non-linear process optimization using simulated annealing and tabu search

Screening of topologies developed by hierarchical heuristic procedures can be carried out by comparing their optimal performance. In this work we will be exploiting mono-objective process optimization using two algorithms, simulated annealing and tabu search, and four different objective functions: two of the net present value type, one of them including environmental costs and two of the global potential impact type. The hydrodealkylation of toluene to produce benzene was used as case study, considering five topologies with different complexities mainly obtained by including or not liquid recycling and heat integration. The performance of the algorithms together with the objective functions was observed, analyzed and discussed from various perspectives: average deviation of results for each algorithm, capacity for producing high purity product, screening of topologies, objective functions robustness in screening of topologies, trade-offs between economic and environmental type objective functions and variability of optimum solutions.

Entropy diversity in multi-objective particle swarm optimization

Multi-objective particle swarm optimization (MOPSO) is a search algorithm based on social behavior. Most of the existing multi-objective particle swarm optimization schemes are based on Pareto optimality and aim to obtain a representative non-dominated Pareto front for a given problem. Several approaches have been proposed to study the convergence and performance of the algorithm, particularly by accessing the final results. In the present paper, a different approach is proposed, by using Shannon entropy to analyzethe MOPSO dynamics along the algorithm execution. The results indicate that Shannon entropy can be used as an indicator of diversity and convergence for MOPSO problems.

Optimization of QuEChERS procedure coupled to GC-ECD for organochlorine pesticide determination in carrot samples

An optimised version of the Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) method for simultaneous determination of 14 organochlorine pesticides in carrots was developed using gas chromatography coupled with electron-capture detector (GC-ECD) and confirmation by gas chromatography tandem mass spectrometry (GC-MS/MS). A citrate-buffered version of QuEChERS was applied for the extraction of the organochlorine pesticides, and for the extract clean-up, primary secondary amine, octadecyl-bonded silica (C18), magnesium sulphate (MgSO4) and graphitized carbon black were used as sorbents. The GC-ECD determination of the target compounds was achieved in less than 20 min. The limits of detection were below the EUmaximum residue limits (MRLs) for carrots, 10–50 μg kg−1, while the limit of quantification did exceed 10 μg kg−1 for hexachlorobenzene (HCB). The introduction of a sonication step was shown to improve the recoveries. The overall average recoveries in carrots, at the four tested levels (60, 80, 100 and 140 μg kg−1), ranged from 66 to 111% with relative standard deviations in the range of 2– 15 % (n03) for all analytes, with the exception of HCB. The method has been applied to the analysis of 21 carrot samples from different Portuguese regions, and β-HCH was the pesticide most frequently found, with concentrations oscillating between less than the limit of quantification to 14.6 μg kg−1. Only one sample had a pesticide residue (β-HCH) above the MRL, 14.6 μg kg−1. This methodology combines the advantages of both QuEChERS and GC-ECD, producing a very rapid, sensitive and reliable procedure which can be applied in routine analytical laboratories.

Optimization of a microplate-based assay to assess esterase activity in the alga Pseudokirchneriella subcapitata

The present work describes the optimization of a short-term assay, based on the inhibition of the esterase activity of the alga Pseudokirchneriella subcapitata, in a microplate format. The optimization of the staining procedure showed that the incubation of the algal cells with 20 μmolL−1 fluorescein diacetate (FDA) for 40 min allowed discrimination between metabolic active and inactive cells. The shortterm assay was tested using Cu as toxicant. For this purpose, algal cells, in the exponential or stationary phase of growth, were exposed to the heavy metal in growing conditions. After 3 or 6 h, cells were subsequently stained with FDA, using the optimized procedure. For Cu, the 3- and 6-h EC50 values, based on the inhibition of the esterase activity of algal cells in the exponential phase of growth, were 209 and 130 μg L−1, respectively. P. subcapitata cells, in the stationary phase of growth, displayed higher effective concentration values than those observed in the exponential phase. The 3- and 6-h EC50 values for Cu, for cells in the stationary phase, were 443 and 268 μgL−1, respectively. This short-term microplate assay showed to be a rapid endpoint for testing toxicity using the alga P. subcapitata. The small volume required, the simplicity of the assay (no washing steps), and the automatic reading of the fluorescence make the assay particularly well suited for the evaluation of the toxicity of a high number of environmental samples.

Demand response management in power systems using a particle swarm optimization approach

Competitive electricity markets have arisen as a result of power-sector restructuration and power-system deregulation. The players participating in competitive electricity markets must define strategies and make decisions using all the available information and business opportunities.

Application-specific modified particle swarm optimization for energy resource scheduling considering vehicle-to-grid

This paper presents a modified Particle Swarm Optimization (PSO) methodology to solve the problem of energy resources management with high penetration of distributed generation and Electric Vehicles (EVs) with gridable capability (V2G). The objective of the day-ahead scheduling problem in this work is to minimize operation costs, namely energy costs, regarding he management of these resources in the smart grid context. The modifications applied to the PSO aimed to improve its adequacy to solve the mentioned problem. The proposed Application Specific Modified Particle Swarm Optimization (ASMPSO) includes an intelligent mechanism to adjust velocity limits during the search process, as well as self-parameterization of PSO parameters making it more user-independent. It presents better robustness and convergence characteristics compared with the tested PSO variants as well as better constraint handling. This enables its use for addressing real world large-scale problems in much shorter times than the deterministic methods, providing system operators with adequate decision support and achieving efficient resource scheduling, even when a significant number of alternative scenarios should be considered. The paper includes two realistic case studies with different penetration of gridable vehicles (1000 and 2000). The proposed methodology is about 2600 times faster than Mixed-Integer Non-Linear Programming (MINLP) reference technique, reducing the time required from 25 h to 36 s for the scenario with 2000 vehicles, with about one percent of difference in the objective function cost value.

Convex optimization framework for intermediate deadline assignment in soft and hard real-time distributed systems

It is generally challenging to determine end-to-end delays of applications for maximizing the aggregate system utility subject to timing constraints. Many practical approaches suggest the use of intermediate deadline of tasks in order to control and upper-bound their end-to-end delays. This paper proposes a unified framework for different time-sensitive, global optimization problems, and solves them in a distributed manner using Lagrangian duality. The framework uses global viewpoints to assign intermediate deadlines, taking resource contention among tasks into consideration. For soft real-time tasks, the proposed framework effectively addresses the deadline assignment problem while maximizing the aggregate quality of service. For hard real-time tasks, we show that existing heuristic solutions to the deadline assignment problem can be incorporated into the proposed framework, enriching their mathematical interpretation.

Internet based VRS Code Positioning

Absolute positioning – the real time satellite based positioning technique that relies solely on global navigation satellite systems – lacks accuracy for several real time application domains. To provide increased positioning quality, ground or satellite based augmentation systems can be devised, depending on the extent of the area to cover. The underlying technique – multiple reference station differential positioning – can, in the case of ground systems, be further enhanced through the implementation of the virtual reference station concept. Our approach is a ground based system made of a small-sized network of three stations where the concept of virtual reference station was implemented. The stations provide code pseudorange corrections, which are combined using a measurement domain approach inversely proportional to the distance from source station to rover. All data links are established trough the Internet.

Spinning reserve and emission unit commitment through stochastic optimization

This paper proposes a stochastic mixed-integer linear approach to deal with a short-term unit commitment problem with uncertainty on a deregulated electricity market that includes day-ahead bidding and bilateral contracts. The proposed approach considers the typically operation constraints on the thermal units and a spinning reserve. The uncertainty is due to the electricity prices, which are modeled by a scenario set, allowing an acceptable computation. Moreover, emission allowances are considered in a manner to allow for the consideration of environmental constraints. A case study to illustrate the usefulness of the proposed approach is presented and an assessment of the cost for the spinning reserve is obtained by a comparison between the situation with and without spinning reserve.

Shannon information and power law analysis of the chromosome code

This paper studies the information content of the chromosomes of twenty-three species. Several statistics considering different number of bases for alphabet character encoding are derived. Based on the resulting histograms, word delimiters and character relative frequencies are identified. The knowledge of this data allows moving along each chromosome while evaluating the flow of characters and words. The resulting flux of information is captured by means of Shannon entropy. The results are explored in the perspective of power law relationships allowing a quantitative evaluation of the DNA of the species.

Shannon entropy analysis of the genome code

This paper studies the chromosome information of twenty five species, namely, mammals, fishes, birds, insects, nematodes, fungus, and one plant. A quantifying scheme inspired in the state space representation of dynamical systems is formulated. Based on this algorithm, the information of each chromosome is converted into a bidimensional distribution. The plots are then analyzed and characterized by means of Shannon entropy. The large volume of information is integrated by averaging the lengths and entropy quantities of each species. The results can be easily visualized revealing quantitative global genomic information.

On the use of code mobility mechanisms in real-time systems

Applications with soft real-time requirements can benefit from code mobility mechanisms, as long as those mechanisms support the timing and Quality of Service requirements of applications. In this paper, a generic model for code mobility mechanisms is presented. The proposed model gives system designers the necessary tools to perform a statistical timing analysis on the execution of the mobility mechanisms that can be used to determine the impact of code mobility in distributed real-time applications.

Online robust optimization framework for QoS guarantees in distributed soft real-time systems

In distributed soft real-time systems, maximizing the aggregate quality-of-service (QoS) is a typical system-wide goal, and addressing the problem through distributed optimization is challenging. Subtasks are subject to unpredictable failures in many practical environments, and this makes the problem much harder. In this paper, we present a robust optimization framework for maximizing the aggregate QoS in the presence of random failures. We introduce the notion of K-failure to bound the effect of random failures on schedulability. Using this notion we define the concept of K-robustness that quantifies the degree of robustness on QoS guarantee in a probabilistic sense. The parameter K helps to tradeoff achievable QoS versus robustness. The proposed robust framework produces optimal solutions through distributed computations on the basis of Lagrangian duality, and we present some implementation techniques. Our simulation results show that the proposed framework can probabilistically guarantee sub-optimal QoS which remains feasible even in the presence of random failures.

Value of a lower limb immobilization device for SPECT/CT image fusion optimization

The foot and the ankle are small structures commonly affected by disorders, and their complex anatomy represent significant diagnostic challenges. SPECT/CT Image fusion can provide missing anatomical and bone structure information to functional imaging, which is particularly useful to increase diagnosis certainty of bone pathology. However, due to SPECT acquisition duration, patient’s involuntary movements may lead to misalignment between SPECT and CT images. Patient motion can be reduced using a dedicated patient support. We aimed at designing an ankle and foot immobilizing device and measuring its efficacy at improving image fusion. Methods: We enrolled 20 patients undergoing distal lower-limb SPECT/CT of the ankle and the foot with and without a foot holder. The misalignment between SPECT and CT images was computed by manually measuring 14 fiducial markers chosen among anatomical landmarks also visible on bone scintigraphy. Analysis of variance was performed for statistical analysis. Results: The obtained absolute average difference without and with support was 5.1±5.2 mm (mean±SD) and 3.1±2.7 mm, respectively, which is significant (p<0.001). Conclusion: The introduction of the foot holder significantly decreases misalignment between SPECT and CT images, which may have clinical influence in the precise localization of foot and ankle pathology.