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PubMed2006

A practical approach to optimization and validation of a HPLC assay for analysis of polyribosyl-ribitol phosphate in complex combination vaccines.

The use of multi-factor statistical experimental design methodology minimized the vaccine material and laboratory resources required for optimization and validation of an HPLC assay for quantitation of depolymerized and total PRP. Components of the assay selected for optimization were adjuvant dissolution, ultracentrifuge conditions including ultracentrifuge model, sample diluent, mobile phase and column oven temperature. Previous experience has shown these components of the assay to be most troublesome and therefore required optimization prior to validation. Specificity, linearity, precision, accuracy and ruggedness were confirmed through a validation of the optimized assay. The validation also established the assay to be stability indicating, by showing that changes to the integrity of the PRP-OMPC conjugate could be detected.

Chromatography, High Pressure Liquid↗

PubMed2004

Formulation optimization of water-in-oil-water multiple emulsion for intestinal insulin delivery.

Formulation optimization of the water-in-oil-in-water multiple emulsion incorporating insulin was performed based on statistical methods such as the orthogonal experimental design and the response surface evaluation. As model formulations, 16 types of emulsions were prepared according to the orthogonal experimental design. To optimize the formulation, the influence of causal factors such as amounts of gelatin, insulin, oleic acid (OA) and the volume ratio of the outer aqueous phase to total and agitation time of the second emulsification process on individual characteristics of the emulsion, such as inner droplet size, viscosity, stability and pharmacological effect, was evaluated first. Based on the analysis of ANOVA, it was concluded that the droplet size of the emulsion was influenced by the volume ratio of the outer aqueous phase significantly. The viscosity of the emulsion was affected by these causal factors and their interactions; however, the most predominant contribution of all causal factors was the volume ratio of the outer aqueous phase. Similarly, one of the most important characteristics in the design of the formulation, stability, was affected by the causal factors. With regard to the hypoglycemic effect, the most influential factor was the content of OA in the emulsion. By means of a novel optimization technique involving a multivariate spline interpolation (MSI), formulation optimization was performed with respect to pharmacological effect and stability, and the optimum formulation with a desirable pharmacological effect and high stability was successfully estimated.

Animals↗

PubMed2004

Wavefront-optimized ablation profiles: theoretical background.

PURPOSE: To describe a method for calculating wavefront-optimized ablation profiles to precompensate for the spherical aberration and higher-order astigmatism induced by myopic, hyperopic, and astigmatic corneal laser corrections. SETTING: IROC-Institut für Refraktive und Ophthalmo-Chirurgie, and Institute for Biomedical Engineering, Swiss Federal Institute of Technology, Zürich, Switzerland. METHODS: The basic ablation profile for myopic, hyperopic, and astigmatic correction is derived from the 2nd-order Zernike representation of wavefront aberrations. Including 4th-order spherical aberration and higher-order astigmatism in the theoretical calculation of the ablation profile allows precompensation for the expected amount of higher-order aberrations (HOAs). The shapes of wavefront-optimized ablation profiles are compared with the shapes of "classic" ablation profiles for myopic and astigmatic corrections. RESULTS: The introduction of precompensating spherical aberration and higher-order astigmatism leads to a more aspheric ablation profile with a significant increase in ablation depth (up to 35%) in the midperiphery of the optical zone. The central ablation depth remains unchanged in the myopic correction but increases by 3% in cylinder correction. CONCLUSIONS: Wavefront-optimized ablation profiles provide a simple method to precompensate for the expected 4th-order spherical aberration and higher-order astigmatism in the average eye. Further clinical studies must be performed to prove the theoretical results; demonstrate the reduction in HOAs; and predict safety, predictability, and stability of wavefront-optimized ablation profiles.

Algorithms↗

PubMed2006

An inexact optimization approach for river water-quality management.

A previously developed fuzzy waste load allocation model (FWLAM) for a river system is extended to address uncertainty involved in fixing the membership functions for the fuzzy goals of the pollution control agency (PCA) and the dischargers using the concept of grey systems. The model provides flexibility for the PCA and the dischargers to specify their goals independently, as the parameters for membership functions are considered as interval grey numbers instead of deterministic real numbers. An inexact or a grey fuzzy optimization model is developed in a multiobjective framework, to maximize the width of the interval valued fractional removal levels for providing latitude in decision-making and to minimize the width of the goal fulfillment level for reducing the system uncertainty. The concept of an acceptability index for order relation between two partially or fully overlapping intervals is used to get a deterministic equivalent of the grey fuzzy optimization model developed. The improvement of the optimal solutions over a previously developed grey fuzzy waste load allocation model (GFWLAM) is shown through an application to a hypothetical river system. The fuzzy multiobjective optimization and fuzzy goal programming techniques are used to solve the deterministic equivalent of the GFWLAM.

Algorithms↗

PubMed2005

Identifying optimal agricultural countermeasure strategies for a hypothetical contamination scenario using the strategy model.

A spatially implemented model designed to assist the identification of optimal countermeasure strategies for radioactively contaminated regions is described. Collective and individual ingestion doses for people within the affected area are estimated together with collective exported ingestion dose. A range of countermeasures are incorporated within the model, and environmental restrictions have been included as appropriate. The model evaluates the effectiveness of a given combination of countermeasures through a cost function which balances the benefit obtained through the reduction in dose with the cost of implementation. The optimal countermeasure strategy is the combination of individual countermeasures (and when and where they are implemented) which gives the lowest value of the cost function. The model outputs should not be considered as definitive solutions, rather as interactive inputs to the decision making process. As a demonstration the model has been applied to a hypothetical scenario in Cumbria (UK). This scenario considered a published nuclear power plant accident scenario with a total deposition of 1.7x10(14), 1.2x10(13), 2.8x10(10) and 5.3x10(9)Bq for Cs-137, Sr-90, Pu-239/240 and Am-241, respectively. The model predicts that if no remediation measures were implemented the resulting collective dose would be approximately 36 000 person-Sv (predominantly from 137Cs) over a 10-year period post-deposition. The optimal countermeasure strategy is predicted to avert approximately 33 000 person-Sv at a cost of approximately 160 million pounds. The optimal strategy comprises a mixture of ploughing, AFCF (ammonium-ferric hexacyano-ferrate) administration, potassium fertiliser application, clean feeding of livestock and food restrictions. The model recommends specific areas within the contaminated area and time periods where these measures should be implemented.

Agriculture↗

PubMed2006

Biodegradation of dye solution containing Malachite Green: optimization of effective parameters using Taguchi method.

In this paper, optimization of biological decolorization of synthetic dye solution containing Malachite Green was investigated. The effect of temperature, initial pH of the solution, type of algae, dye concentration and time of the reaction was studied and optimized using Taguchi method. Sixteen experiments were required to study the effect of parameters on biodegradation of the dye. Each of experiments was repeated three times to calculate signal/noise (S/N). Our results showed that initial pH of the solution was the most effective parameter in comparison with others and the basic pH was favorable. In this study, we also optimized the experimental parameters and chose the best condition by determination effective factors. Based on the S/N ratio, the optimized conditions for dye removal were temperature 25 degrees C, initial pH 10, dye concentration 5 ppm, algae type Chlorella and time 2.5h. The stability and efficiency of Chlorella sp. in long-term repetitive operations were also examined.

Animals↗

PubMed2004

Optimization of intracellular cytokine staining for the quantitation of antigen-specific CD4+ T cell responses in rhesus macaques.

Standard proliferation assays used for analysis of CD4+ T cell function have significant shortcomings, including limited sensitivity, lack of truly quantitative readouts and significant variability. We have optimized an intracellular cytokine staining (ICS) assay in rhesus macaques which allows us to identify virus-specific CD4+ T cells at the single-cell level with high sensitivity while reducing background staining to a minimum. A variety of parameters were tested to determine the optimal experimental conditions necessary for the detection of antigen-specific CD4+ T cells in macaques. Central to our optimized protocol was the addition of cross-linked costimulatory anti-CD28 and anti-CD49d Mabs, a modification which resulted in up to threefold enhancement of the frequency of TNF-alpha-secreting CD4+ T cells following superantigen- or antigen-specific stimulation. The ICS protocol was also optimized with respect to antigen concentration and duration of antigenic stimulation. These modifications resulted in a convenient and highly reproducible assay with intra- and inter-assay variability of less than 10%. Although cryopreservation of PBMC generally led to a 40% to 80% decrease in the frequency of antigen-specific CD4+ T cells detected by ICS using stimulation with viral proteins, the use of overlapping peptide pools minimized the effects of cryopreservation on ICS responses. The use of more sensitive techniques such as ICS permits delineation of antigen-specific cells at the single cell level and should provide new insights into pathogen-specific immune responses in the rhesus macaque model.

Animals↗

PubMed2005

MEKC-LIF of gamma-amino butyric acid in microdialysate: systematic optimization of the separation conditions by factorial analysis.

Micellar electrokinetic chromatography allows the efficient separation of biogenic amines and amino acids in biological samples. Analytes of interest, sample composition, and sample matrix may vary between studies, which necessitates optimization of separations to meet the requirements and conditions of an experiment. Factorial analysis is an efficient tool to accomplish this type of optimization involving multiple interacting factors. The present study describes an optimization procedure for separation of the inhibitory neurotransmitter GABA utilizing capillary electrophoresis with laser induced fluorescence detection. Standards labeled with the flourogenic reagent 3-(2-furoyl)quinoline-2 carboxaldehyde were separated with varying concentrations of borate buffer, beta-cyclodextrin, sodium dodecyl sulfate and pH. The optimized separation method had a correlation coefficient between concentration of GABA and fluorescent signal of 0.98, and was linear in the desired concentration range of 25 nM-10 microM. Glutamic acid, aspartic acid and taurine were also quantified using this separation. When applied to microdialysate collected from the region of the suprachiasmatic nucleus, this separation was able to measure daily variations in GABA levels. The factorial design experiment has proven to be a useful tool, allowing adjustments in the separation of neurotransmitters based on individual requirements.

Chromatography, Micellar Electrokinetic Capillary↗

PubMed2006

Optimization of GC-MS conditions based on resolution and stability of analytes for simultaneous determination of nine sesquiterpenoids in three species of Curcuma rhizomes.

GC-MS is a powerful tool for analysis of volatile oil, and resolutions of analytes were exclusively used as marker for optimization of the conditions. However, volatile oil usually contains heat labile components which may degrade and result in wrong results during GC analysis. In present study, based on both resolutions and stabilities of 11 sesquiterpenoids, GC-MS conditions were optimized for simultaneously quantitative determination of nine compounds including beta-elemene, curzerene, curcumol, isocurcumenol, germacrone, curdione, curcumenol, neocurdione and curcumenone in Ezhu. However, the other two compounds, i.e. furanodienone and furanodiene, were still thermal sensitive and not available for GC analysis. The results showed that both resolutions and stabilities of analytes should be considered for optimization of GC conditions because the properties of most components in volatile oil are unknown. Under optimum conditions, a capillary column (30 m x 0.25 mm i.d.) coated with 0.25 microm film 5% phenyl methyl siloxane was used for separation. Pulsed splitless inlet with temperature of 190 degrees C was selected for sample injection (0.2 microl). The calibration curves of nine sesquiterpenoids showed good linearity (r2>0.9989) within test ranges. The optimized method showed good repeatability for quantification of these nine components in Ezhu with intra- and inter-day variations of less than 1.42% and 2.79%, respectively. The validated method was successfully applied to quantify 9 sesquiterpenoids in 18 samples of 3 species of Curcuma used as Ezhu.

Calibration↗

PubMed2007

Construction of an autologous tissue-engineered venous conduit from bone marrow-derived vascular cells: optimization of cell harvest and seeding techniques.

BACKGROUND: Currently available vascular grafts for pediatric cardiovascular operations are limited by their inability to grow. Tissue-engineering techniques can be used to create vascular grafts with the potential for repair, remodeling, and growth. This study demonstrates the feasibility of constructing an autologous tissue-engineered venous conduit from bone marrow-derived vascular cells (BMVCs) in the ovine animal model. METHODS: Ovine mononuclear cells were isolated from the bone marrow, cultured in endothelial growth medium, and characterized with immunocytochemistry. Biodegradable tubular scaffolds were constructed from polyglycolic acid mesh coated with a copolymer of poly[epsilon-caprolactone-L-lactide]. Scaffolds were seeded at various cell concentrations and incubation times to optimize seeding conditions for the construction of an autologous venous conduit. Using optimized conditions, 6 tissue-engineered vascular grafts were implanted as inferior vena cava interposition grafts in juvenile lambs. Grafts were assessed for patency at days 1 to 30 postoperatively and explanted for histological and immunohistochemical analysis. RESULTS: A mixed cell population of BMVCs consisting of smooth muscle cells and endothelial cells was cultured from ovine sternal bone marrow. A seeding concentration of 2 x 10(6) cells/cm2 and 7 days of postseeding incubation were optimal for creating a confluent cellular layer on the polyglycolic acid/poly[epsilon-caprolactone-L-lactide]) scaffold. Grafts were explanted up to 4 weeks postoperatively. All grafts were patent without evidence of thrombosis. Histological evaluation of the explanted grafts demonstrated neo-endothelialization. Graft wall was composed of neo-tissue made up of residual polymer matrix, mesenchymal cells, and extracellular matrix without evidence of calcification. CONCLUSIONS: Bone marrow-derived vascular cells, containing endothelial and smooth muscle cells, can be isolated and cultured from ovine sternal bone marrow and used as a cell source for vascular tissue engineering. Our optimized techniques for BMVC harvest and seeding onto biodegradable scaffolds can be used for studying autologous tissue-engineered vascular grafts in the ovine animal model.

Animals↗

PubMedDate not supplied

Role of statistical symmetries in sensory coding: an optimal scale invariant code for vision.

The visual system is the most studied sensory pathway, which is partly because visual stimuli have rather intuitive properties. There are reasons to think that the underlying principle ruling coding, however, is the same for vision and any other type of sensory signal, namely the code has to satisfy some notion of optimality--understood as minimum redundancy or as maximum transmitted information. Given the huge variability of natural stimuli, it would seem that attaining an optimal code is almost impossible; however, regularities and symmetries in the stimuli can be used to simplify the task: symmetries allow predicting one part of a stimulus from another, that is, they imply a structured type of redundancy. Optimal coding can only be achieved once the intrinsic symmetries of natural scenes are understood and used to the best performance of the neural encoder. In this paper, we review the concepts of optimal coding and discuss the known redundancies and symmetries that visual scenes have. We discuss in depth the only approach which implements the three of them known so far: translational invariance, scale invariance and multiscaling. Not surprisingly, the resulting code possesses features observed in real visual systems in mammals.

Animals↗

PubMed2004

Dynamic optimization of host defense, immune memory, and post-infection pathogen levels in mammals.

When attacked by pathogens, higher vertebrates produce specific immune cells that fight against them. We here studied the host's optimal schedule of specific immune cell production. The damage caused by the pathogen increases with the pathogen amount in the host integrated over time. On the other hand, there is also a cost incurred by the production of specific immune cells, not only in terms of the energy needed to produce and maintain the cells, but also with respect to damages sustained by the host's body as a result of immune activity. The optimal strategy of the host is the one that minimizes the total cost, defined as a weighted sum of the damage caused by pathogens and the costs caused by the specific immune cells. The problem is solved by using Pontryagin's maximum principle and dynamic programming. The optimal defense schedule is typically as follows: In the initial phase after infection, immune cells are produced at the fastest possible rate. The amount of pathogen increases temporarily but is eventually suppressed. When the pathogen amount is suppressed to a sufficiently low level, the immune cell number decreases and converges to a low steady level, which is maintained by alternately switching between fastest production and no production. We examine the effect of time delay required to have fully active immune cells by comparing cases with different number of rate limiting steps before producing immune cells. We examine the effect of the duration of time (time delay) required before full-scale production of active immune cells by comparing cases with different numbers of rate-limiting steps before immune-cell production. We also discuss the role of immune memory based on the results of the optimal immune reaction.

Animals↗

PubMed2004

Mate choice when males are in patches: optimal strategies and good rules of thumb.

In standard mate-choice models, females encounter males sequentially and decide whether to inspect the quality of another male or to accept a male already inspected. What changes when males are clumped in patches and there is a significant cost to travel between patches? We use stochastic dynamic programming to derive optimum strategies under various assumptions. With zero costs to returning to a male in the current patch, the optimal strategy accepts males above a quality threshold which is constant whenever one or more males in the patch remain uninspected; this threshold drops when inspecting the last male in the patch, so returns may occur only then and are never to a male in a previously inspected patch. With non-zero within-patch return costs, such a two-threshold rule still performs extremely well, but a more gradual decline in acceptance threshold is optimal. Inability to return at all need not decrease performance by much. The acceptance threshold should also decline if it gets harder to discover the last males in a patch. Optimal strategies become more complex when mean male quality varies systematically between patches or years, and females estimate this in a Bayesian manner through inspecting male qualities. It can then be optimal to switch patch before inspecting all males on a patch, or, exceptionally, to return to an earlier patch. We compare performance of various rules of thumb in these environments and in ones without a patch structure. A two-threshold rule performs excellently, as do various simplifications of it. The best-of-N rule outperforms threshold rules only in non-patchy environments with between-year quality variation. The cutoff rule performs poorly.

Animals↗

PubMed2005

A mathematical consideration for the optimal shell change of hermit crab.

Shell of the adult hermit crab has some important roles for its fitness. In the same time, the shell size often limits the body growth of its owner. To grow the body size larger, the individual must change the shell to another larger shell. If the individual cannot get another larger one, the individual has to suppress the body size growth as the occupied shell size allows. Growth suppression would result in the lower fitness. With a simple mathematical model, we consider the criterion about whether the individual should try to change the shell or not in order to get the higher fitness. We show that the optimality of a shell change behavior has a relation with the body size and the season length for the shell change. They also affect the optimal timing for the shell change. It is implied that the probability of the success in a shell change and the cost for the shell change behavior do not affect the optimal timing for the shell change at all but significantly do the optimality of the behavioral choice.

Animals↗

PubMed2005

Optimized arterial trees supplying hollow organs.

Computer models of arterial trees can be generated from optimization principles using the algorithm of constrained constructive optimization (CCO). Up to now this algorithm could handle only tissue areas of convex shape, without concavities. CCO is now generalized to cope also with non-convex organ shapes, possibly featuring external as well as internal concavities. This allows the modeling of a much larger class of interesting real arterial systems. The concept of a generalized domain-potential was developed to represent arbitrary non-convex shapes mathematically and incorporate them as boundary conditions to optimization. Domain-potentials may be derived from analytical representations as well as from finite element triangulations obtained from organ images. To demonstrate the feasibility of the concept, the optimized growth of an arterial tree model is confined to some part of an elliptical shell, representing the free wall of the left ventricle of the heart.

Algorithms↗

PubMed2004

Development and validation of evolutionary algorithm software as an optimization tool for biological and environmental applications.

A flexible, extendable tool for the optimization of (micro)biological processes and protocols using evolutionary algorithms was developed. It has been tested using three different theoretical optimization problems: 2 two-dimensional problems, one with three maxima and one with five maxima and a river autopurification optimization problem with boundary conditions. For each problem, different evolutionary parameter settings were used for the optimization. For each combination of evolutionary parameters, 15 generations were run 20 times. It has been shown that in all cases, the evolutionary algorithm gave rise to valuable results. Generally, the algorithms were able to detect the more stable sub-maximum even if there existed less stable maxima. The latter is, from a practical point of view, generally more desired. The most important factors influencing the convergence process were the parameter value randomization rate and distribution. The developed software, described in this work, is available for free.

Algorithms↗

PubMed2005

Optimal imaging parameters for fiber-orientation estimation in diffusion MRI.

This study uses Monte Carlo simulations to investigate the optimal value of the diffusion weighting factor b for estimating white-matter fiber orientations using diffusion MRI with a standard spherical sampling scheme. We devise an algorithm for determining the optimal echo time, pulse width, and pulse separation in the pulsed-gradient spin-echo sequence for a specific value of b. The Monte Carlo simulations provide an estimate of the optimal value of b for recovering one and two fiber orientations. We show that the optimum is largely independent of the noise level in the measurements and the number of gradient directions and that the optimum depends only weakly on the diffusion anisotropy, the maximum gradient strength, and the spin-spin relaxation time. The optimum depends strongly on the mean diffusivity. In brain tissue, the optima we estimate are in the ranges [0.7, 1.0] x 10(9) s m(-2) and [2.2, 2.8] x 10(9) s m(-2) for the one- and two-fiber cases, respectively. The best b for estimating the fractional anisotropy is slightly higher than for estimating fiber directions in the one-fiber case and slightly lower in the two-fiber case. To estimate Tr(D) in the one-fiber case, the optimal setting is higher still. Simulations suggest that a ratio of high to low b measurements of 5 to 1 is a good compromise for measuring fiber directions and size and shape indices.

Algorithms↗

PubMed2004

Intranasal administration of an Escherichia coli-expressed codon-optimized rotavirus VP6 protein induces protection in mice.

We are developing rotavirus vaccines based on the VP6 protein of the human G1P[8] [corrected] [J. Virol. 73 (1999) 7574] CJN strain of rotavirus. One prototype candidate consisting of MBP::VP6::His6, a chimeric protein of maltose-binding protein, VP6 and hexahistidine, was expressed mainly as truncated polypeptides in Escherichia coli BL21(DE3) cells. A possible reason for this extensive truncation is the high frequencies of rare bacterial codons within the rotavirus VP6 gene. Expression of truncated recombinant VP6 was found to be reduced, and expression of complete VP6 protein was simultaneously increased, when the protein was expressed in Rosetta(DE3)pLacI E. coli cells that contain increased amounts of transfer RNAs for a selection of rare codons. The same observation was made when a synthetic codon-optimized CJN-VP6 gene was expressed in E. coli BL21 or Rosetta cells. To increase protein recovery, recombinant E. coli cells were treated with 8M urea. Denatured, full-length MBP::VP6::His6 protein was then purified and used for intranasal vaccination of BALB/c mice (2 doses administered with E. coli heat-labile toxin LT(R192G) as adjuvant). Following oral challenge with the G3P[16] [corrected] [J. Virol. 76 (2002) 560] EDIM strain of murine rotavirus, protection levels against fecal rotavirus shedding were comparable (P>0.05) between groups of mice immunized with denatured codon-optimized or native (not codon-optimized) immunogen with values ranging from 87 to 99%. These protection levels were also comparable to those found after immunization with non-denatured CJN VP6. Thus, expression of complete rotavirus VP6 protein was greatly enhanced by codon optimization, and the protection elicited was not affected by denaturation of recombinant VP6.

Administration, Intranasal↗