Development of Cationic Solid Lipid Nanoparticles with Factorial Design-Based Studies for Topical Administration of Doxorubicin

Topical chemotherapy using doxorubicin, a powerful anticancer drug, can be used as an alternative with reduced systemic toxicity when treating skin cancer. The aim of the present work was to use factorial design-based studies to develop cationic solid lipid nanoparticles containing doxorubicin; further investigations into the influence of these particles on the drug's cytotoxicity and cellular uptake in B16F10 murine melanoma cells were performed. A 3(2) full factorial design was applied for two different lipid phases; one phase used stearic acid and the other used a 1:2 mixture of stearic acid and glyceryl behenate. The two factors investigated included the ratio between the lipid and the water phase and the ratio between the surfactant (poloxamer) and the co-surfactant (cetylpyridinium chloride). It was observed that the studied factors did not affect the mean diameter or the polydispersity of the obtained nanoparticles; however, they did significantly affect the zeta potential values. Optimised formulations with particle sizes ranging from 251 to 306 nm and positive zeta potentials were selected for doxorubicin incorporation. High entrapment efficiencies were achieved (97%) in formulations with higher amounts of stearic acid, suggesting that cationic charges on doxorubicin molecules may interact with the negative charges in stearic acid. Melanoma culture cell experiments showed that cationic solid lipid nanoparticles without drug were not cytotoxic to melanoma cells. The encapsulation of doxorubicin significantly increased cytotoxicity, indicating the potential of these nanoparticles for the treatment of skin cancer.

An optimization method for conceptual design in engineering systems based on high order singular value decomposition

Se presenta un nuevo método de diseño conceptual en Ingeniería Aeronáutica basado el uso de modelos reducidos, también llamados modelos sustitutos (‘surrogates’). Los ingredientes de la función objetivo se calculan para cada indiviudo mediante la utilización de modelos sustitutos asociados a las distintas disciplinas técnicas que se construyen mediante definiciones de descomposición en valores singulares de alto orden (HOSVD) e interpolaciones unidimensionales. Estos modelos sustitutos se obtienen a partir de un número limitado de cálculos CFD. Los modelos sustitutos pueden combinarse, bien con un método de optimización global de tipo algoritmo genético, o con un método local de tipo gradiente. El método resultate es flexible a la par que mucho más eficiente, computacionalmente hablando, que los modelos convencionales basados en el cálculo directo de la función objetivo, especialmente si aparecen un gran número de parámetros de diseño y/o de modelado. El método se ilustra considerando una versión simplificada del diseño conceptual de un avión. Abstract An optimization method for conceptual design in Aeronautics is presented that is based on the use of surrogate models. The various ingredients in the target function are calculated for each individual using surrogates of the associated technical disciplines that are constructed via high order singular value decomposition and one dimensional interpolation. These surrogates result from a limited number of CFD calculated snapshots. The surrogates are combined with an optimization method, which can be either a global optimization method such as a genetic algorithm or a local optimization method, such as a gradient-like method. The resulting method is both flexible and much more computationally efficient than the conventional method based on direct calculation of the target function, especially if a large number of free design parameters and/or tunablemodeling parameters are present. The method is illustrated considering a simplified version of the conceptual design of an aircraft empennage.

Comments and remarks over classic linear loop-gain method for oscillator design and analysis. New proposed method based on NDF/RRT

Abstract. This paper describes a new and original method for designing oscillators based on the Normalized Determinant Function (NDF) and Return Relations (RRT)- Firstly, a review of the loop-gain method will be performed. The loop-gain method pros, cons and some examples for exploring wrong solutions provided by this method will be shown. This method produces in some cases wrong solutions because some necessary conditions have not been fulfilled. The required necessary conditions to assure a right solution will be described. The necessity of using the NDF or the Transpose Return Relations (RRT), which are related with the True Loop-Gain, to test the additional conditions will be demonstrated. To conclude this paper, the steps for oscillator design and analysis, using the proposed NDF/RRj method, will be presented. The loop-gain wrong solutions will be compared with the NDF/RRj and the accuracy of this method to estimate the oscillation frequency and QL will be demonstrated. Some additional examples of plane reference oscillators (Z/Y/T), will be added and they will be analyzed with the new NDF/RRj proposed method, even these oscillators cannot be analyzed using the classic loop gain method.

Application of the Equivalent Static Analysis method to the design of a steel frame structure with added viscous dampers

All the structures designed by engineers are vulnerable to natural disasters including floods and earthquakes. The energy released during strong ground motions should be dissipated by structural elements. Before 1990’s, this energy was expected to be dissipated through the beams and columns which at the same time were a part of gravity-load-resisting system. However, the main disadvantage of this idea was that gravity-resisting-frame was not repairable. Hence, during 1990’s, the idea of designing passive energy dissipation systems, including dampers, emerged. At the beginning, main problem was lack of guidelines for passive energy dissipation systems. Although till 2000 many guidelines and procedures where published, yet most of them were based on complicated analysis which was not so convenient for engineers and practitioners. In order to solve this problem recently some alternative design methods are proposed including 1. Lopez Garcia (2001) simple procedure for optimal damper configuration in MDOF structures 2. Christopoulos and Filiatrault (2006) trial and error procedure 3. Silvestri et al. (2010) Five-Step Method. 4. Palermo et al. (2015) Direct Five-Step Method. 5. Palermo et al. (2016) Simplified Equivalent Static Analysis (ESA). In this study, effectiveness and differences between last three alternative methods have been evaluated.

Comparison of the implicit enumeration method and the branch-and-bound method for logical design

Bibliography: p. 91-94.

Fractional factorial experiment designs for factors at two levels.

Bibliography: p. 4.

A structural method for the design of fault tolerant distributed control systems

Distributed digital control systems provide alternatives to conventional, centralised digital control systems. Typically, a modern distributed control system will comprise a multi-processor or network of processors, a communications network, an associated set of sensors and actuators, and the systems and applications software. This thesis addresses the problem of how to design robust decentralised control systems, such as those used to control event-driven, real-time processes in time-critical environments. Emphasis is placed on studying the dynamical behaviour of a system and identifying ways of partitioning the system so that it may be controlled in a distributed manner. A structural partitioning technique is adopted which makes use of natural physical sub-processes in the system, which are then mapped into the software processes to control the system. However, communications are required between the processes because of the disjoint nature of the distributed (i.e. partitioned) state of the physical system. The structural partitioning technique, and recent developments in the theory of potential controllability and observability of a system, are the basis for the design of controllers. In particular, the method is used to derive a decentralised estimate of the state vector for a continuous-time system. The work is also extended to derive a distributed estimate for a discrete-time system. Emphasis is also given to the role of communications in the distributed control of processes and to the partitioning technique necessary to design distributed and decentralised systems with resilient structures. A method is presented for the systematic identification of necessary communications for distributed control. It is also shwon that the structural partitions can be used directly in the design of software fault tolerant concurrent controllers. In particular, the structural partition can be used to identify the boundary of the conversation which can be used to protect a specific part of the system. In addition, for certain classes of system, the partitions can be used to identify processes which may be dynamically reconfigured in the event of a fault. These methods should be of use in the design of robust distributed systems.

Improved production of industrially relevant recombinant proteins: application of multi-factorial design of experiments and scalable process modelling

The work described in this thesis focuses on the use of a design-of-experiments approach in a multi-well mini-bioreactor to enable the rapid establishments of high yielding production phase conditions in yeast, which is an increasingly popular host system in both academic and industrial laboratories. Using green fluorescent protein secreted from the yeast, Pichia pastoris, a scalable predictive model of protein yield per cell was derived from 13 sets of conditions each with three factors (temperature, pH and dissolved oxygen) at 3 levels and was directly transferable to a 7 L bioreactor. This was in clear contrast to the situation in shake flasks, where the process parameters cannot be tightly controlled. By further optimisating both the accumulation of cell density in batch and improving the fed-batch induction regime, additional yield improvement was found to be additive to the per cell yield of the model. A separate study also demonstrated that improving biomass improved product yield in a second yeast species, Saccharomyces cerevisiae. Investigations of cell wall hydrophobicity in high cell density P. pastoris cultures indicated that cell wall hydrophobin (protein) compositional changes with growth phase becoming more hydrophobic in log growth than in lag or stationary phases. This is possibly due to an increased occurrence of proteins associated with cell division. Finally, the modelling approach was validated in mammalian cells, showing its flexibility and robustness. In summary, the strategy presented in this thesis has the benefit of reducing process development time in recombinant protein production, directly from bench to bioreactor.

Analysing Conjoint Analysis Data by a Random Coefficient Regression Model

2000 Mathematics Subject Classification: 62J12, 62K15, 91B42, 62H99.

Extração, caracterização e avaliação bioativa do extrato de Arrabidaea chica

The use of plants for medicinal purposes is ancient, with widespread application in medicinal drugs. Although plants are promising sources for the discovery of new molecules of pharmacological interest, estimates show that only 17% of them have been studied for their possible use in medicine. Thus, biodiversity of Brazilian flora represents an immense potential for economic use by the pharmaceutical industry. The plant Arrabidaea chica, popularly known as “pariri”, is common in the Amazon region, and it is assigned several medicinal properties. The leaves of this plant are rich in anthocyanins, which are phenolic compounds with high antioxidant power. Antioxidant compounds play a vital role in the prevention of neurological and cardiovascular diseases, cancer and diabetes, among others. Within the anthocyanins found in Arrabidaea chica, stands out Carajurin (6,7-dihydroxy-5,4’- dimethoxy-flavilium), which is the major pigment encountered in this plant. The present work aimed to study on supercritical extraction and conventional extraction (solid-liquid extraction) in leaves of Arrabidaea chica, evaluating the efficiency of the extractive processes, antioxidant activity and quantification of Carajurin contained in the extracts. Supercritical extraction used CO2 as solvent with addition of co-solvent (ethanol/water mixture) and were conducted by the dynamic method in a fixed bed extractor. The trials followed a 24-1 fractional factorial design, the dependent variables were: process yield, concentration of Carajurin and antioxidant activity; and independent variables were: pressure, temperature, concentration of co-solvent (v/v) and concentration of water in the co-solvent mixture (v/v). Yields (mass of dry extract/mass of raw material used) obtained from supercritical extraction ranged from 15.1% to 32%, and the best result was obtained at 250 bar and 40 °C, co-solvent concentration equal to 30% and concentration of water in the co-solvent mixture equal to 50%. Through statistical analysis, it was found that the concentration of co-solvent revealed significant effect on the yield. Yields obtained from conventional extractions were of 8.1% (water) and 5.5% (ethanol). Through HPLC (High-performance liquid chromatography) analysis, Carajurin was quantified in all the extracts and concentration values (Carajurin mass/mass of dry extract) ranged between 1% and 2.21% for supercritical extraction. For conventional extraction, Carajurin was not detected in the aqueous extract, while the ethanol extract showed Carajurin content of 7.04%, and therefore, more selective in Carajurin than the supercritical extraction. Evaluation of antioxidant power (radical 2,2-diphenyl-1-picrylhydrazyl – DPPH – sequestration method) of the supercritical extracts resulted in EC50 values (effective concentration which neutralizes 50% of free radicals) ranged from 38.34 to 86.13 μg/mL, while conventional extraction resulted in EC50 values of 167.34 (water) and 42.58 (ethanol) μg/mL. As for the quantification of total phenolic content (Folin-Ciocalteau analysis) of the supercritical extracts resulted in values ranged from 48.93 and 88.62 mg GAE/g extract (GAE = Gallic Acid Equivalents), while solid-liquid extraction resulted in values of 37.63 (water) and 80.54 (ethanol) mg GAE/g extract. The good antioxidant activity cannot be attributed solely to the presence of Carajurin, but also the existence of other compounds and antioxidants in Arrabidaea chica. By optimizing the experimental design, it was possible to identify the experiment that presented the best result considering the four dependent variables together. This experiment was performed under the following conditions: pressure of 200 bar, temperature of 40 °C, co-solvent concentration equal to 30% and concentration of water in the co-solvent mixture equal to 30%. It is concluded that, within the studied range, it is possible to purchase the optimum result using milder operating conditions, which implies lower costs and greater ease of operation.

Optimization of microwave-assisted extration of phenolic compounds from tomao by full factorial design coupled with response surface methodology

Tomato (Lycopersicon esculentum Mill.), apart from being a functional food rich in carotenoids, vitamins and minerals, is also an important source of phenolic compounds [1 ,2]. As antioxidants, these functional molecules play an important role in the prevention of human pathologies and have many applications in nutraceutical, pharmaceutical and cosmeceutical industries. Therefore, the recovery of added-value phenolic compounds from natural sources, such as tomato surplus or industrial by-products, is highly desirable. Herein, the microwave-assisted extraction of the main phenolic acids and flavonoids from tomato was optimized. A S-Ieve! full factorial Box-Behnken design was implemented and response surface methodology used for analysis. The extraction time (0-20 min), temperature (60-180 "C), ethanol percentage (0-100%), solidlliquid ratio (5-45 g/L) and microwave power (0-400 W) were studied as independent variables. The phenolic profile of the studied tomato variety was initially characterized by HPLC-DAD-ESIIMS [2]. Then, the effect of the different extraction conditions, as defined by the used experimental design, on the target compounds was monitored by HPLC-DAD, using their UV spectra and retention time for identification and a series of calibrations based on external standards for quantification. The proposed model was successfully implemented and statistically validated. The microwave power had no effect on the extraction process. Comparing with the optimal extraction conditions for flavonoids, which demanded a short processing time (2 min), a low temperature (60 "C) and solidlliquid ratio (5 g/L), and pure ethanol, phenolic acids required a longer processing time ( 4.38 min), a higher temperature (145.6 •c) and solidlliquid ratio (45 g/L), and water as extraction solvent. Additionally, the studied tomato variety was highlighted as a source of added-value phenolic acids and flavonoids.

Encapsulation of Local Anesthetic Bupivacaine in Biodegradable Poly(DL-lactide-co-glycolide) Nanospheres: Factorial Design, Characterization and Cytotoxicity Studies

Local anesthetic agents cause temporary blockade of nerve impulses productiong insensitivity to painful stimuli in the area supplied by that nerve. Bupivacaine (BVC) is an amide-type local anesthetic widely used in surgery and obstetrics for sustained peripheral and central nerve blockade. in this study, we prepared and characterized nanosphere formulations containing BVC. To achieve these goals, BVC loaded poly(DL-lactide-co-glycolide) (PLGA) nanospheres (NS) were prepared by nanopreciptation and characterized with regard to size distribution, drug loading and cytotoxicity assays. The 2(3-1) factorial experimental design was used to study the influence of three different independent variables on nanoparticle drug loading. BVC was assayed by HPLC, the particle size and zeta potential were determined by dynamic light scattering. BVC was determined using a combined ultrafiltration-centrifugation technique. The results of optimized formulations showed a narrow size distribution with a polydispersivity of 0.05%, an average diameter of 236.7 +/- 2.6 nm and the zeta potential -2.93 +/- 1,10 mV. In toxicity studies with fibroblast 3T3 cells, BVC loaded-PLGA-NS increased cell viability, in comparison with the effect produced by free BVC. In this way, BVC-loaded PLGA-NS decreased BVC toxicity. The development of BVC formulations in carriers such as nanospheres could offer the possibility of controlling drug delivery in biological systems, prolonging the anesthetic effect and reducing toxicity.

Partial microwave-assisted wet digestion of animal tissue using a baby-bottle sterilizer for analyte determination by inductively coupled plasma optical emission spectrometry

A procedure for partial digestion of bovine tissue is proposed using polytetrafluoroethylene (PTFE) microvessels inside a baby-bottle sterilizer under microwave radiation for multi-element determination by inductively coupled plasma optical emission spectrometry (ICP OES). Samples were directly weighed in laboratory-made polytetrafluoroethylene vessels. Nitric acid and hydrogen peroxide were added to the uncovered vessels, which were positioned inside the baby-bottle sterilizer, containing 500 mL of water. The hydrogen peroxide volume was fixed at 100 mu L The system was placed in a domestic microwave oven and partial digestion was carried out for the determination of Ca, Cu, Fe. Mg, Mn and Zn by inductively coupled plasma optical emission spectrometry. The single-vessel approach was used in the entire procedure, to minimize contamination in trace analysis. Better recoveries and lower residual carbon content (RCC) levels were obtained under the conditions established through a 2(4-1) fractional factorial design: 650 W microwave power, 7 min digestion time, 50 mu L nitric acid and 50 mg sample mass. The digestion efficiency was ascertained according to the residual carbon content determined by inductively coupled plasma optical emission spectrometry. The accuracy of the proposed procedure was checked against two certified reference materials. (C) 2009 Elsevier B.V. All rights reserved.

Extractive Fermentation of Clavulanic Acid by Streptomyces DAUFPE 3060 Using Aqueous Two-Phase System

The influence of four variables, specifically PEG molar mass (400, 1,000, and 8,000 g/mol), concentrations of PEG and phosphate salts (15, 20, and 25% for both), and agitation intensity (110, 150, and 200 rpm), on clavulanic acid (CA) extraction by extractive fermentation with PEG/phosphate salts aqueous two-phase system was investigated in shaken flasks using a 2(4-1)-fractional factorial design. After selection of the two most significant variables (agitation intensity and PEG molar mass), an optimization study conducted according to a 2(2)-central composite design revealed that 25% PEG 8,000 g/mol and phosphate salts at 240 rpm (run 6) were the best conditions for the extractive fermentation, leading to the best results in terms of partition coefficient (k = 8.2), yield of CA in the PEG-rich phase (eta(T) = 93%) and productivity (P = 5.3 mg/Lh). As a first attempt to make a scale-up of these results, the effectiveness of the extractive fermentation was then checked in a bench-scale bioreactor under conditions as close as possible to the optimum ones determined in flasks. The highest CA concentration obtained in the PEG-rich phase (691 mg/L) was 30% higher than in flasks, thus demonstrating the potential of such a new process, integrating the production and extraction steps, as a promising, low-cost tool to obtain high yields of this and similar products. (C) 2010 American Institute of Chemical Engineers Biotechnol. Prog., 27: 95-103, 2011