The cytotoxicity of methacryloxylethyl cetyl ammonium chloride, a cationic antibacterial monomer, is related to oxidative stress and the intrinsic mitochondrial apoptotic pathway

Antibacterial monomers incorporated in dentin bonding systems may have toxic effects on the pulp. Thus, the cytotoxicity of antibacterial monomers and its underlying mechanisms must be elucidated to improve the safety of antibacterial monomer application. The influence of an antibacterial monomer, methacryloxylethyl cetyl ammonium chloride (DMAE-CB), on the vitality of L929 mouse fibroblasts was tested using MTT assay. Cell cycle progression was studied using flow cytometry. Production of intracellular reactive oxygen species (ROS) after DMAE-CB treatment was measured using 2,7-dichlorodihydrofluorescein diacetate staining and flow cytometry analysis. Loss of mitochondrial membrane potential, disturbance of Bcl-2 and Bax expression, as well as release of cytochrome C were also measured using flow cytometry analysis or Western blot to explore the possible involvement of the mitochondrial-related apoptotic pathway. DMAE-CB elicited cell death in a dose-dependent manner and more than 50% of cells were killed after treatment with 30 µM of the monomer. Both necrosis and apoptosis were observed. DMAE-CB also induced G1- and G2-phase arrest. Increased levels of intracellular ROS were observed after 1 h and this overproduction was further enhanced by 6-h treatment with the monomer. DMAE-CB may cause apoptosis by disturbing the expression of Bcl-2 and Bax, reducing the mitochondrial potential and inducing release of cytochrome C. Taken together, these findings suggest that the toxicity of the antibacterial monomer DMAE-CB is associated with ROS production, mitochondrial dysfunction, cell cycle disturbance, and cell apoptosis/necrosis.

The presence of qacA/B gene in Brazilian methicillin-resistant Staphylococcus aureus

A total of 74 methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from three government hospitals in 2002 and 2003 were examined concerning the distribution of qacA/B gene, which is the determinant of resistance to quaternary ammonium compounds largely employed in hospital disinfection. By polymerase chain reaction the qacA/B gene was found in 80% of the isolates, which is a significant result considering it is the first time that qacA/B gene is being reported for Brazilian MRSA strains and it is presented at a high rate.

Estudo de adsorção de compostos sulfurados utilizando um diesel comercial dopado com benzotiofeno e dibenzotiofeno

Desulphurization process by adsorption was studied employing a commercial diesel dooped with 1000 mg/L of benzothiophene and dibenzothiophene. The adsorbents materials employed were three types of activated alumina (acid, basic and neutral). For comparison, adsorption process was made also using oxidized diesel sample. The results showed that the adsorbents were selective for sulphur compounds removal from fuels. The contact time have influence in adsorption process achieving 80% of removal for not oxidized dibenzothiophene. The three studied alumina types showed similar behavior and a greater selective in dibenzothiophene adsorption than benzothiophene. Dibenzothiophene removal is more effective in samples not oxidized, whereas the benzothiophene was almost totally removed in oxidized sample.

Evaluation of the efficacy of commercial sanitizers against adhered and planktonic cells of Listeria monocytogenes and Salmonella spp.

Antimicrobial activities of two commercial disinfectants, alone or combined with heat, against three Salmonella strains and three Listeria monocytogenes strains were studied. The efficacy of disinfectants against planktonic bacteria and bacteria attached to three food contact industrial surfaces (stainless steel, polytetraflourethylene, and rubber) was investigated. The tests were conducted using the sanitizer (quaternary ammonium compounds, and alquyldiethylenediamineglycine and di-alquyldiamineethylglycine) concentrations recommended by the manufacturers, and concentrations twice and four times higher than those values. The recommended concentrations were not effective to kill bacteria, especially when they were attached to surfaces. Concentrations of disinfectants twice and four times higher than those recommended were needed to fully eliminate planktonic bacteria. These same sanitizer concentrations were not sufficient to remove attached bacteria. To remove them from the surfaces, a treatment with recommended concentrations in combination with heat was needed. Our results indicate that these two pathogenic bacteria could survive common sanitation programs used in the food industry.

Adsorption capacity of phenolic compounds onto cellulose and xylan

The interaction between three phenolic compounds (catechin, caffeic acid and ferulic acid) onto two dietary fibres (cellulose and xylan) has been evaluated to inquire possible interferences on the biodisponibility of phenolic compounds. The adsorption kinetics were performed using solutions containing 100 mg/L of phenolic compounds during a contact time ranging between 10 and 120 minutes at pH 2.0, 4.5, and 7.0. After the kinetics, isotherms were obtained using phenolic compounds concentration ranging between 10 and 80 mg/L during 60 minutes, at pH 2.0 and 7.0 and temperature of 36 °C. Results indicate that adsorbed quantities mainly changed in function of pH, however the maximum adsorption was only of 0.978 mg of caffeic acid/g of xylan at pH 2 and after 60 min. Redlich-Peterson model were able to predict the adsorption isotherms of all phenolic compounds onto cellulose, except for caffeic acid at pH 7.0. The low adsorption capacities observed suggest that both dietary fibres are unable to compromise the biodisponibility of phenolic compounds, especially in the small intestine, where they are partially absorbed.

Application of mathematical models for the prediction of adsorption isotherms in solid mixture for mango powder refreshment

Solid mixtures for refreshment are already totally integrated to the Brazilian consumers' daily routine, because of their quick preparation method, yield and reasonable price - quite lower if compared to 'ready-to-drink' products or products for prompt consumption, what makes them economically more accessible to low-income populations. Within such a context, the aim of this work was to evaluate the physicochemical and mineral composition, as well as the hygroscopic behavior of four different brands of solid mixture for mango refreshment. The BET, GAB, Oswim and Henderson mathematical models were built through the adjustment of experimental data to the isotherms of adsorption. Results from the physiochemical evaluation showed that the solid mixtures for refreshments are considerable sources of ascorbic acid and reductor sugar; and regarding mineral compounds, they are significant sources of calcium, sodium and potassium. It was also verified that the solid mixtures for refreshments of the four studied brands are considered highly hygroscopic.

Initial performance of pineapple and utilization of rock phosphate applied in combination with organic compounds to leaf axils

Rock phosphates have low solubility in water, but good solubility in acid. The use of organic compounds together with these phosphorus sources applied to the basal leaf axils of pineapple can increase the solubility of this phosfate source and increase the P availability to the crop. A greenhouse experiment was conducted using Araxá rock phosphate (10 g) in combination or not with solutions containing increasing concentrations of humic acids (0 to 40 mmol L-1 of carbon), with or without citric acid (0.005 mmol L-1), applied to basal leaf axils of pineapple cv. Pérola. Growth and nutritional characteristics of aerial plant parts were assessed. Growth rates of aerial parts and N, P, K, Ca and Mg contents increased curvilinearly with increasing concentration of carbon in the form of humic acids. Maximum values were found for the concentration of 9.3 mmol L-1 of carbon combined with 0.005 mmol L-1 of citric acid and natural phosphate.

Health promoting and sensory properties of phenolic compounds in food

Phenolic compounds have been extensively studied in recent years. The presence of these compounds in various foods has been associated with sensory and health promoting properties. These products from the secondary metabolism of plants act as defense mechanisms against environmental stress and attack by other organisms. They are divided into different classes according to their chemical structures. The objective of this study was to describe the different classes of phenolic compounds, the main food sources and factors of variation, besides methods for the identification and quantification commonly used to analyze these compounds. Moreover, the role of phenolic compounds in scavenging oxidative stress and the techniques of in vitro antioxidant evaluation are discussed. In vivo studies to evaluate the biological effects of these compounds and their impact on chronic disease prevention are presented as well. Finally, it was discussed the role of these compounds on the sensory quality of foods.

Influence of strawberry jam color and phenolic compounds on acceptance during storage

The objective of this study was to evaluate the influence of the color and phenolic compounds of strawberry jam on acceptance during storage. Jams were processed, stored for 120 days and evaluated monthly for chromatic characteristics, total phenolic compounds, total anthocyanins (ANT), total ellagic acid (TEA), flavonoids and free ellagic acid (FEA), and sensory acceptance as well. Data were submitted to analysis of variance (ANOVA) and the means were compared by the Least Significant Difference (LSD). Cluster Analysis and Partial Least Square Regression (PLS) were performed to investigate the relationships between instrumental data and acceptance. Contents of ANT, TEA and redness decreased during storage. Other chemical characteristics and sensory acceptance showed a nonlinear behavior. Higher acceptance was observed after 60 days, suggesting a trend of quality improvement followed by decline to the initial levels. The same trend was observed for lightness, non-pigment flavonoids and FEA. According to PLS map, for consumers in cluster 2, acceptance was associated to jams at 60 days and to luminosity, FEA, and non-pigment flavonoids. For cluster 1, a positive association between flavor liking, jam at initial storage, and the contents of TEA and ANT was indicated. Jams at 120 days were positively associated to hue and negatively associated to color liking, for cluster 1. Color and texture were positively correlated to overall liking for cluster 2, whereas for cluster 1, overall acceptance seemed to be more associated to flavor liking. Changes in color and phenolic compounds slightly influenced the acceptance of strawberry jams, but in different ways for consumers clusters.

Cumulative mortality of Aedes aegypti larvae treated with compounds

OBJECTIVE To evaluate the larvicidal activity of Azadirachta indica, Melaleuca alternifolia, carapa guianensis essential oils and fermented extract of Carica papaya against Aedes aegypti (Linnaeus, 1762) (Diptera: Culicidae). METHODS The larvicide test was performed in triplicate with 300 larvae for each experimental group using the third larval stage, which were exposed for 24h. The groups were: positive control with industrial larvicide (BTI) in concentrations of 0.37 ppm (PC1) and 0.06 ppm (PC2); treated with compounds of essential oils and fermented extract, 50.0% concentration (G1); treated with compounds of essential oils and fermented extract, 25.0% concentration (G2); treated with compounds of essential oils and fermented extract, 12.5% concentration (G3); and negative control group using water (NC1) and using dimethyl (NC2). The larvae were monitored every 60 min using direct visualization. RESULTS No mortality occurred in experimental groups NC1 and NC2 in the 24h exposure period, whereas there was 100% mortality in the PC1 and PC2 groups compared to NC1 and NC2. Mortality rates of 65.0%, 50.0% and 78.0% were observed in the groups G1, G2 and G3 respectively, compared with NC1 and NC2. CONCLUSIONS The association between three essential oils from Azadirachta indica, Melaleuca alternifolia, Carapa guianensis and fermented extract of Carica papaya was efficient at all concentrations. Therefore, it can be used in Aedes aegypti Liverpool third larvae stage control programs.

Effect of bioactive compounds extracted from euphorbious plants on hematological and biochemical parameters of Channa punctatus

Channa punctatus was exposed to four different concentrations of Rutin, Taraxerol and Apigenin. Changes in some hematological parameters of Channa punctatus were assessed to determine the influence of these compounds on test fish. Fish were exposed to sublethal concentrations (80% of LC50 of 24h) of these compounds for one week. Control fish were also administered for one week. Thereafter, blood samples were obtained from the control and experimental fish. Blood was assayed for selected hematological parameters (hematocrit, hemoglobin, red blood cell count, white blood cell count total plasma protein and plasma glucose concentration). The derived hematological indices of mean corpuscular hemoglobin concentration (MCHC), mean corpuscular hemoglobin (MCH) and mean corpuscular volume (MCV) were calculated. Sublethal concentrations of these compounds caused a dose dependent decrease in hemoglobin values coupled with a decrease in hematocrit values and red blood cell counts are an obvious indication of anemia. The total white blood cell counts and the differential white blood cell counts were decreased except for the lymphocytes, where there was a slight increase. Plasma protein and glucose were also lower in exposed fish when compared with control. The hematological indices MCH, MCHC, MCV were also lowered. The result from this study reveals high mortality rate and deleterious consequences on the health of fish subjected to acute exposure of Rutin, Taraxerol and Apigenin and therefore, should not be used directly in aquaculture without having the proper knowledge.

Leishmanicidal activity and cytotoxicity of compounds from two Annonacea species cultivated in Northeastern Brazil

INTRODUCTION: Visceral leishmaniasis is endemic in 88 countries, with a total of 12 million people infected and 350 million at risk. In the search for new leishmanicidal agents, alkaloids and acetogenins isolated from leaves of Annona squamosa and seeds of Annona muricata were tested against promastigote and amastigote forms of Leishmania chagasi. METHODS: Methanol-water (80:20) extracts of A. squamosa leaves and A. muricata seeds were extracted with 10% phosphoric acid and organic solvents to obtain the alkaloid and acetogenin-rich extracts. These extracts were chromatographed on a silica gel column and eluted with a mixture of several solvents in crescent order of polarity. The compounds were identified by spectroscopic analysis. The isolated compounds were tested against Leishmania chagasi, which is responsible for American visceral leishmaniasis, using the MTT test assay. The cytotoxicity assay was evaluated for all isolated compounds, and for this assay, RAW 264.7 cells were used. RESULTS: O-methylarmepavine, a benzylisoquinolinic alkaloid, and a C37 trihydroxy adjacent bistetrahydrofuran acetogenin were isolated from A. squamosa, while two acetogenins, annonacinone and corossolone, were isolated from A. muricata. Against promastigotes, the alkaloid showed an IC50 of 23.3 µg/mL, and the acetogenins showed an IC50 ranging from 25.9 to 37.6 µg/mL; in the amastigote assay, the IC50 values ranged from 13.5 to 28.7 µg/mL. The cytotoxicity assay showed results ranging from 43.5 to 79.9 µg/mL. CONCLUSIONS: These results characterize A. squamosa and A. muricata as potential sources of leishmanicidal agents. Plants from Annonaceae are rich sources of natural compounds and an important tool in the search for new leishmanicidal therapies.

Avaliação da capacidade de fixação de fósforo pelo solo

Phosphate "fixation" is the convertion of soluble into insoluble phosphate in the soil. There are many factors conditioning phosphate fixation by soil such as reactions originating less soluble compounds (phosphates of iron, aluminum, calcium, magnesium, etc.), PO4-3 adsorption by the colloidal fraction of soils, PO4-3 absorption by the soil microflora, etc. Certain soils of the state of São Paulo (Brazil) are relatively rich in both iron and aluminum oxides. PO4-3 fixation, using P31 and P32 has been verified by researchers, specially with "Terra Roxa". The known methods for fixation evaluation are conventional as this depends on phosphate solution concentration, pH, time of contact between soil and solution, relation of sample weight to solution volume, shaking time, etc. In this experiment, the following conventional method was used: 4 g of soil were shaken for 15 minutes at 30-40 rpm, in 300 ml Erlenmeyer flask in a Wagner shaking machine, together with 100 ml of 0,03 normal phosphate solution (being 0,01 normal as PO4-3 contributed by H8PO4 and 0,02 normal as PO4-3 from KH2PO4). After shaking it was set aside for 24 hours and then filtered. Phosphate was determined in a suitable aliquot of both the original solution (blank) and the soil extract, by the vanadomolibidic-phosphoric acid method. From phosphate concentration in the blank minus phosphate concentration in the soil stract the rate of fixation by 100 g of soil was calculated. The data obtained show that "Terra Roxa" and "Terra Roxa Misturada" have a fairly high PO4-3 fixation capacity, varying from 10 to 24 milliequivalents of PO4-3 per 100 g of soil.

Process of fixation, imobilization and mineralization of ammonium in soil using N-15

The organic and inorganic forms of soil nitrogen and how they participate in the process of fixation, immobilization and mineralization of ammonium in soils were evaluated, after different periods of incubaton, utilizing two soils, a Lithic Haplustoll and a Typic Eutrorthox. The results obtained permit to suggest that : 1) The method for determination of the ammonium fixing capacity based on the extraction with 2N KC1, is considered to be subject to interferences of other soil fractions capable of retaining ammonium. 2) The increase in exchangeable ammonium content is related to the decrease in amino acids and hydrolyzable ammonium. 3) The immobilization and mineralization processes are still held under mil microbial. The forms more affected by this condition are amino acids and hydrolyzable ammonium.

Antitumor activity of chemical modified natural compounds

Search of new activity substances starting from chemotherapeutic agents, continously appears in international literature. Perhaps this search has been done more frequently in the field of anti-tumor chemotherapy on account of the unsuccess in saving advanced stage patients. The new point in this matter during the last decade was computer aid in planning more rational drugs. In near future "the accessibility of supercomputers and emergence of computer net systems, willopen new avenues to rational drug design" (Portoghese, P. S. J. Med. Chem. 1989, 32, 1). Unknown pharmacological active compounds synthetized by plants can be found even without this eletronic devices, as tradicional medicine has pointed out in many contries, and give rise to a new drug. These compounds used as found in nature or after chemical modifications have produced successful experimental medicaments as FAA, "flavone acetic acid" with good results as inibitors of slow growing animal tumors currently in preclinical evaluation for human treatment. In this lecture some international contributions in the field of chemical modified compounds as antineoplasic drugs will be examined, particularly those done by Brazilian researches.