Therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia using Fe2O3 nanoparticles

This study aimed to investigate the therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia (MFH) using Fe2O3 nanoparticles. Hepatocarcinoma SMMC-7721 cells cultured in vitro were treated with ferrofluid containing Fe2O3 nanoparticles and irradiated with an alternating radio frequency magnetic field. The influence of the treatment on the cells was examined by inverted microscopy, MTT and flow cytometry. To study the therapeutic mechanism of the Fe2O3 MFH, Hsp70, Bax, Bcl-2 and p53 were detected by immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). It was shown that Fe2O3 MFH could cause cellular necrosis, induce cellular apoptosis, and significantly inhibit cellular growth, all of which appeared to be dependent on the concentration of the Fe2O3 nanoparticles. Immunocytochemistry results showed that MFH could induce high expression of Hsp70 and Bax, decrease the expression of mutant p53, and had little effect on Bcl-2. RT-PCR indicated that Hsp70 expression was high in the early stage of MFH (<24 h) and became low or absent after 24 h of MFH treatment. It can be concluded that Fe2O3 MFH significantly inhibited the proliferation of in vitro cultured liver cancer cells (SMMC-7721), induced cell apoptosis and arrested the cell cycle at the G2/M phase. Fe2O3 MFH can induce high Hsp70 expression at an early stage, enhance the expression of Bax, and decrease the expression of mutant p53, which promotes the apoptosis of tumor cells.

THE EFFECTIVE DIFFUSIVITY OF CLOVE(Eugenia caryophyllus) ESSENTIAL OIL IN PRESSURIZED CO2

The effective diffusivity of clove essential oil in subcritical liquid CO2 was estimated. The experimental apparatus employed was a fixed-bed extractor. The fixed bed was formed with grounded (mesh -32 + 65) and compacted clove buds which were considered a solid element. The effective diffusion coefficient was evaluated by fitting the experimental concentration profile to the unsteady state mass balance equation for unidirectional diffusion in a finite solid medium. The diffusion coefficient was related to the concentration of oil in the solid by an exponential function. The estimated values of the effective diffusion coefficient varied from 3.64 to 5.22x10-10 m2/s. The average relative errors were lower than 3.1%.

COMPARISON OF GINGER (Zingiber officiale Roscoe) OLEORESIN OBTAINED WITH ETHANOL AND ISOPROPANOL WITH THAT OBTAINED WITH PRESSURIZED CO2

Ginger (Zingiber officinale Roscoe) belongs to the Zingiberacea family. It is a spice of great commercial importance. In this work ginger oleoresin was obtained with ethanol, isopropanol and liquid carbon dioxide. The chemical compositions of the extract were compared with each other. All oleoresin samples had monoterpenes and sesquiterpenes. Carboxylic acids were found in organic solvent extracts for an extraction time of 2 hours. The component responsible the for pungent characteristic of the oleoresin, gingerois, were detected in samples obtained with organic solvent for extraction times of 6 hours and in samples obtained with CO2 liquid for extraction times of 2 hours.

NOVAS APLICAÇÕES DE SISTEMAS SFE "HOME MADE".: I. PLANTAS MEDICINAIS BRASILEIRAS

Neste trabalho, apresenta-se uma visão geral da evolução do uso da SFE (extração com fluido supercrítico) na análise de plantas medicinais brasileiras dentro das pesquisas desenvolvidas pelo Laboratório de Cromatografia do IQSC-USP. Utilizou-se como fluido extrator o CO2, puro ou modificado com solventes de polaridade baixa a alta (pentano, solventes halogenados, álcoois, água), para a extração seletiva de princípios ativos de Maytenus aquifolium ("espinheira santa") e Mikania glomerata ("guaco"), respectivamente contendo triterpenos e cumarina. Os extratos obtidos por SFE foram analisados por CGAR (cromatografia gasosa de alta resolução), CLAE-DAD (cromatografia líquida de alta eficiência com detector" photodiodearray") ou CCD (cromatografia de camada delgada). A SFE forneceu resultados similares ou melhores do que os obtidos com métodos convencionais de extração (Soxhlet, maceração, etc), indicando o potencial uso da SFE para análise e/ou produção de preparações contendo estas plantas medicinais.

NOVAS APLICAÇÕES DE SISTEMAS SFE" HOME MADE".: II. PLANTAS DA AMÉRICA DO SUL

O Laboratório de Cromatografia do IQSC-USP tem desenvolvido pesquisas em conjunto com outros grupos de pesquisa do Brasil e da América Latina, visando a futura implantação da SFE (extração com fluido supercrítico) nestes laboratórios. No presente trabalho, apresenta-se alguns dos resultados obtidos na cooperação com pesquisadores do Chile e da Venezuela, direcionados para a SFE de substâncias de potencial interesse econômico, princípios ativos ou substâncias tóxicas: triglicerídeos, ácidos graxos, flavonóides, alcalóides, etc. Foram estudadas plantas nativas do Chile e da Venezuela, algumas delas nunca estudadas por métodos fitoquímicos convencionais e pertencentes a várias famílias vegetais: Leguminosae, Chrysobalanaceae, Annonnaceae, etc. O uso da SFE em conjunto com CGAR-EM (cromatografia gasosa de alta resolução - espectrometria de massas) ou CLAE-DAD (cromatografia líquida de alta eficiência com detector" photodiodearray") permitiu a identificação de algumas substâncias presentes em baixa concentração, a partir de pequenas quantidades de material vegetal; estes dados são de grande importância para a caracterização química das espécies estudadas.

NOVAS APLICAÇÕES DE SISTEMAS SFE "HOME MADE": III. ENERGIA QUÍMICA E ELÉTRICA

Este trabalho apresenta três aplicações da SFE com diferentes fluídos supercríticos, matrizes e sistemas de extração. Um sistema estático "home made" foi utilizado na pesquisa de fontes alternativas de insumos químicos e energéticos tais como o bagaço de cana-de-açucar e o carvão mineral. Foram usados, respectivamente, o etanol (sem e com catalisador) e o tolueno como fluídos extratores e hidrogênio como gás de pressurização. Na extração dos contaminantes do óleo mineral adsorvidos em bauxita, foi utilizado o dióxido de carbono como fluído extrator em um sistema "home made"/dinâmico. Estudos visando a maximização do rendimento de alguns dos processos também foram realizados, utilizando o planejamento fatorial e a metodologia da superfície de resposta. Os produtos de maior interesse neste trabalho (fração de resinas), obtidos do processo de extração do bagaço de cana-de-açucar e aqueles contaminantes do óleo isolante adsorvidos em bauxita extraídos com fluídos supercríticos foram caracterizados usando cromatografia gasosa de alta resolução com detector de ionização de chama (CGAR - DIC) e cromatografia gasosa de alta resolução com detector seletivo de massas (CGAR - EM) após fracionamento por cromatografia líquida preparativa em 8 frações distintas.

NOVAS APLICAÇÕES DE SISTEMAS SFE "HOME-MADE".: IV. QUALIDADE DOS ALIMENTOS

São apresentadas algumas aplicações de um sistema "Home-Made" para Extração com Fluido Supercrítico (EFS) na análise de resíduos de pesticidas e de óleos essenciais em alimentos. Foram desenvolvidos diversos estudos com os parâmetros: temperatura, pressão, escolha e concentração de modificadores. Os resultados são apresentados para amostras de maracujá (resíduos de pesticidas organofosforados e piretróides), mel (resíduos de pesticidas organoclorados) e laranja (óleo essencial).

NOVAS APLICAÇÕES DE SISTEMAS DE PARA EXTRAÇÃO COM FLUIDO SUPERCRÍTICO (SFE)" HOME-MADE".: VI. CONTROLE DE AGROTÓXICOS EM PRODUTOS ALIMENTÍCIOS

No presente trabalho apresentamos um sistema para extração com fluido supercrítico (SFE)" home-made" utilizado em extrações de resíduos de pesticidas de diversas matrizes. Os pesticidas estudados foram extraídos usando-se como fluido extrator CO2 no estado supercrítico com ou sem modificadores. Os resultados obtidos mostraram-se mais vantajosos do que aqueles obtidos por métodos convencionais de extração.

THE GENERALIZED MAXIMUM LIKELIHOOD METHOD APPLIED TO HIGH PRESSURE PHASE EQUILIBRIUM

The generalized maximum likelihood method was used to determine binary interaction parameters between carbon dioxide and components of orange essential oil. Vapor-liquid equilibrium was modeled with Peng-Robinson and Soave-Redlich-Kwong equations, using a methodology proposed in 1979 by Asselineau, Bogdanic and Vidal. Experimental vapor-liquid equilibrium data on binary mixtures formed with carbon dioxide and compounds usually found in orange essential oil were used to test the model. These systems were chosen to demonstrate that the maximum likelihood method produces binary interaction parameters for cubic equations of state capable of satisfactorily describing phase equilibrium, even for a binary such as ethanol/CO2. Results corroborate that the Peng-Robinson, as well as the Soave-Redlich-Kwong, equation can be used to describe phase equilibrium for the following systems: components of essential oil of orange/CO2.

HIGH PRESSURE PHASE EQUILIBRIUM: PREDICTION OF ESSENTIAL OIL SOLUBILITY

This work describes a method to predict the solubility of essential oils in supercritical carbon dioxide. The method is based on the formulation proposed in 1979 by Asselineau, Bogdanic and Vidal. The Peng-Robinson and Soave-Redlich-Kwong cubic equations of state were used with the van der Waals mixing rules with two interaction parameters. Method validation was accomplished calculating orange essential oil solubility in pressurized carbon dioxide. The solubility of orange essential oil in carbon dioxide calculated at 308.15 K for pressures of 50 to 70 bar varied from 1.7± 0.1 to 3.6± 0.1 mg/g. For same the range of conditions, experimental solubility varied from 1.7± 0.1 to 3.6± 0.1 mg/g. Predicted values were not very sensitive to initial oil composition.

Effect of blanching time on selective mineral elements extraction from the spinach substitute (Tetragonia expansa) commonly used in Brazil

The true spinach (Spinacia oleracea) does not grow well in warm climates and for that reason is not commercialized in Brazil. Instead, a spinach substitute (Tetragonia expansa), originally from New Zealand, is widely used in the country. There is scant information on the mineral profile and none on the soluble mineral fraction of this vegetable. The solubility of a mineral is one of the important factors for its absorption. For this reason, the calcium, magnesium, iron, manganese, copper, zinc, potassium, and sodium soluble fractions in the raw spinach substitute were determined and the effect of blanching times on the solubility of these minerals was investigated. Blanching times of 1, 5, and 15 minutes were employed. The magnesium, manganese, potassium, and sodium soluble fractions increased sizably with shorter blanching time. Longer blanching time (15 minutes) caused large losses of minerals. The soluble mineral fractions can contribute poorly to diet in terms of potassium, magnesium, manganese, and zinc. The spinach substitute cannot be considered a dietary source of calcium, iron and copper due to the insolubility of these minerals in the vegetable, possibly caused by the large oxalate content.

Using Computational Fluid-Dynamics (CFD) for the evaluation of beer pasteurization: effect of orientation of cans

In-package pasteurization is the most used method for beer microbiological stabilization. The search for safer and better quality food has created a need to better understand the processes involved in producing it. However, little is known about the temperature and velocity profiles during the thermal processes of liquid foods in commercial packaging, which results in over-dimensioned processes to guarantee safety, decreasing the sensorial and nutritional characteristics of the product and increasing process costs. Simulations using Computational Fluid-Dynamics (CFD) have been used by various authors to evaluate those processes. The objective of the present paper was to evaluate the effect of packaging orientation in the pasteurization of beer in a commercial aluminum can using CFD. A heating process was simulated at 60 ºC up to 15 PUs (a conventional beer process, in which 1 Pasteurization Unit (PU) is equivalent to 1minute at 60 ºC). The temperature profile and convection current velocity along the process and the variation of the PUs were evaluated in relation to time considering the cans in the conventional, inverted, and horizontal positions. The temperature and velocity profiles were similar to those presented in the literature. The package position did not result in process improvement.

Head Space Solid Phase Micro-Extraction (HS - SPME) of volatile organic compounds produced by Sporidiobolus salmonicolor (CBS 2636)

The aim of the present study was the assessment of volatile organic compounds produced by Sporidiobolus salmonicolor (CBS 2636) using methyl and ethyl ricinoleate, ricinoleic acid and castor oil as precursors. The analysis of the volatile organic compounds was carried out using Head Space Solid Phase Micro-Extraction (HS - SPME). Factorial experimental design was used for investigating extraction conditions, verifying stirring rate (0-400 rpm), temperature (25-60 ºC), extraction time (10-30 minutes), and sample volume (2-3 mL). The identification of volatile organic compounds was carried out by Gas Chromatography with Mass Spectrum Detector (GC/MSD). The conditions that resulted in maximum extraction were: 60 ºC, 10 minutes extraction, no stirring, sample volume of 2.0 mL, and addition of saturated KCl (1:10 v/v). In the bio-production of volatile organic compounds the effect of stirring rate (120-200 rpm), temperature (23-33 ºC), pH (4.0-8.0), precursor concentration (0.02-0.1%), mannitol (0-6%), and asparagine concentration (0-0.2%) was investigated. The bio-production at 28 ºC, 160 rpm, pH 6,0 and with the addition of 0.02% ricinoleic acid to the medium yielded the highest production of VOCs, identified as 1,4-butanediol, 1,2,2-trimethylciclopropilamine, beta-ionone; 2,3-butanodione, pentanal, tetradecane, 2-isononenal, 4-octen-3-one, propanoic acid, and octadecane.

Use of real-time PCR to evaluate two DNA extraction methods from food

The DNA extraction is a critical step in Genetically Modified Organisms analysis based on real-time PCR. In this study, the CTAB and DNeasy methods provided good quality and quantity of DNA from the texturized soy protein, infant formula, and soy milk samples. Concerning the Certified Reference Material consisting of 5% Roundup Ready® soybean, neither method yielded DNA of good quality. However, the dilution test applied in the CTAB extracts showed no interference of inhibitory substances. The PCR efficiencies of lectin target amplification were not statistically different, and the coefficients of correlation (R²) demonstrated high degree of correlation between the copy numbers and the threshold cycle (Ct) values. ANOVA showed suitable adjustment of the regression and absence of significant linear deviations. The efficiencies of the p35S amplification were not statistically different, and all R² values using DNeasy extracts were above 0.98 with no significant linear deviations. Two out of three R² values using CTAB extracts were lower than 0.98, corresponding to lower degree of correlation, and the lack-of-fit test showed significant linear deviation in one run. The comparative analysis of the Ct values for the p35S and lectin targets demonstrated no statistical significant differences between the analytical curves of each target.

Computational fluid dynamics evaluation of liquid food thermal process in a brick shaped package

Food processes must ensure safety and high-quality products for a growing demand consumer creating the need for better knowledge of its unit operations. The Computational Fluid Dynamics (CFD) has been widely used for better understanding the food thermal processes, and it is one of the safest and most frequently used methods for food preservation. However, there is no single study in the literature describing thermal process of liquid foods in a brick shaped package. The present study evaluated such process and the influence of its orientation on the process lethality. It demonstrated the potential of using CFD to evaluate thermal processes of liquid foods and the importance of rheological characterization and convection in thermal processing of liquid foods. It also showed that packaging orientation does not result in different sterilization values during thermal process of the evaluated fluids in the brick shaped package.