EVALUATION OF DIFFERENT TECHNIQUES FOR THE DIFFERENTIATION OF PATHOGENIC AND NON PATHOGENIC STRAINS OF YERSINIA-ENTEROCOLITICA

Different methods and tests have been used to evaluate the pathogenic potential of distinct Y. enterocolitica serotypes and biotypes. We tested a total of 60 Y. enterocolitica strains, being 25 of human origin (serotype O3 biotype 4 and serotype O5 biotype 1); 6 of animal origin (serotype O3 biotype 4); 19 isolated from the environment (serotype O5.27 biotypes 1 and 2); and 8 isolated from food (serotype O5 biotype 1 and serotype 05.27 biotype 1). The methods used were based on plasmid gene expression (autoagglutination, calcium-dependence at 37 degrees C and Congo Red absorption tests), chromosomal gene expression (assays for pyrazinamidase activity, salicin fermentation and esculin hydrolysis), and invasion of HEp-2 cells. All but one of the Y. enterocolitica O3 strains, were found to be potentially pathogenic when submitted to the pyrazinamidase-salicin-esculin tests. In contrast, the results obtained with the assays related to plasmidial gene expression were not so uniform, probably due to plasmid loss. The least homogeneous results were obtained with the HEp-2 cell invasion test. Y. enterocolitica O5 behaved in a uniform manner when tested with the first two groups of tests (based on chromosomal and plasmidial gene expression), but not when tested with the HEp-2 invasion assay. The strains of serotype O5.27 biotype 1 presented a uniform behavior hen submitted to the chromosomic-related tests, showing no pathogenicity. However, they did not provide conclusive results with the tests related to plasmidial gene expression or HEp-2 cell invasion. We conclude that the tests related to chromosomal gene expression (esculin-salicin-pyrazinamidase) are simple and highly effective for the detection of potentially pathogenic Y. enterocolitica isolated from clinical cases.

Isolation and structure elucidation of two new flavonoid glycosides from the infusion of Maytenus aquifolium leaves. Evaluation of the antiulcer activity of the infusion

Droplet countercurrent chromatography and high-performance liquid chromatography fractionation of the aqueous infusion from Maytenus aquifolium Martius leaves afforded two flavonoid tetrasaccharides: quercetin 3-O-alpha-L-rhamnopyranosyl(1-->6)-O-[beta-D-glucopyranosyl(1-->3)-O-alpha-L-rhamnopranosyl( 1-->2)-O-beta-D-galactopyranoside and kaempferol 3-O-alpha-L-rhamnopyranosyl(1-->6)-O-[beta-D-glucopyranosyl( 1-->3)-O-alpha-L-rhamnopyranosy(1-->2-2)-O-beta-D-galactopyranoside. All structures were elucidated by spectroscopic methods. Pharmacological essays of the infusion showed antiulcer activity in rats.

DOES SNATCHING FREQUENCY REALLY INDICATE FOOD INGESTION IN THE NILE TILAPIA

The fitness of the snatching frequency as an indicator of food intake in Nile tilapia finger-lings, Oreochromis niloticus (L), was studied. Five groups of four individuals each were used after a two-day starvation period. The hierarchical rank among individuals in the same group was registered. Food in the form of tiny pellets (ranging from 1.30 to 1.95 mm in diameter) was offered, and the individual snatching frequency was observed during a 20-min period. The animals were then sacrificed for evaluation of stomach contents. It was concluded that snatching frequency is not a good parameter to indicate individual food intake in this species when fed as a group with pellets crushed into tiny particles. This raises a problem for investigations that require evaluation of the cumulative effect of competition on food intake, such as growth or conversion efficiency studies. Furthermore, a very low correlation between snatching frequency and food intake was shown in the third hierarchical rank. It is suggested that the linearity assumed in such hierarchies should be reconsidered.

Evaluation of water and sucrose diffusion coefficients in potato tissue during osmotic concentration

The water and sucrose effective diffusion coefficients behavior were studied in potato tubers immersed in aqueous sucrose solution, 50% (w/,A), at 27 degreesC. Water and sucrose concentration profiles were measured as function of the position for 3, 6 and 12 h of immersion. These were adjusted to a mathematical model for three components that take into account the bulk flow in a shrinking tissue and the concentration dependence of the diffusion coefficients.The binary effective coefficients were an order of magnitude lower than those for pure solutions of sucrose. These coefficients show an unusual concentration dependence. Analysis of these coefficients as functions of the concentration and position demonstrates that, cellular tissue promotes high resistance to diffusion in the tuber and also the elastic contraction of material influences the species diffusion. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

EVALUATION OF DIFFUSION-COEFFICIENTS IN OSMOTIC CONCENTRATION OF BANANAS (MUSA-CAVENDISH LAMBERT)

The effect of time of exposure, solution concentration and temperature on the osmotic concentration of banana (slices of 11 mm thickness) was studied in aqueous sucrose solutions. The selectivity of the cellular tissues was reduced by steam blanching the banana slices before osmotic treatment. Effective diffusion coefficients for the loss of water and the increase in sucrose content were determined according to Fick's Law applied to a two-dimensional body; calculated on the basis of the concentration of various components in the liquid phase impenetrating the fruit. These coefficients revealed values similar to binary diffusion coefficients for pure sucrose solutions.

Evaluation of sample treatment for the simultaneous flow injection hydride generation and determination of As, Bi, Sb and Se by GRAS

A new method was developed for the simultaneous determination of As, Bi, Sb, and Se by flow injection hydride generation graphite furnace atomic absorption spectrometry. An alternative two-step sample treatment procedure was used. The sample was heated (80degreesC) for 10 min in 6 M HCl to reduce Se(VI) to Se(IV), followed by the addition of 1% (m/v) thiourea solution to reduce arsenic and antimony from the pentavalent to the trivalent states.With this procedure, all analytes were converted to their most favorable and sensitive oxidation states to generate the corresponding hydrides. The pre-treated sample solution was then processed in the flow system for in situ trapping and atomization in a graphite tube coated with iridium. The impermanent modifier remained stable up to 300 firings and new coating out significant were possible wit changes in the analytical performance.The accuracy was checked for As, Bi, Sb, and Se determination in water standard reference materials NIST 1640 and 1643d and the results were in agreement with the certified values at a 95% confidence level. Good recoveries (94-104%.) of spiked mineral waters and synthetic As(V), Sb(Ill), mixtures of As(Ill), Sb(V), Se(VI), and Se(IV) were also found. Calculated characteristic masses were 32 mug As, 79 mug Bi, 35 mug Sb, and 130 pg Se, and the corresponding limits of detection were 0.06, 0.16, 0.19, and 0.59 mug L-1, respectively. The repeatability for a typical solution containing 5 mug L-1 As, Bi, Sb, and Se was in the 1-3% range.

Modal representation of three-phase lines applying two transformation matrices: Evaluation of its eigenvectors

The objective of this paper is to show an alternative representation in time domain of a non-transposed three-phase transmission line decomposed in its exact modes by using two transformation matrices. The first matrix is Clarke's matrix that is real, frequency independent, easily represented in computational transient programs (EMTP) and separates the line into Quasi-modes alpha, beta and zero. After that, Quasi-modes a and zero are decomposed into their exact modes by using a modal transformation matrix whose elements can be synthesized in time domain through standard curve-fitting techniques. The main advantage of this alternative representation is to reduce the processing time because a frequency dependent modal transformation matrix of a three-phase line has nine elements to be represented in time domain while a modal transformation matrix of a two-phase line has only four elements. This paper shows modal decomposition process and eigenvectors of a nontransposed three-phase line with a vertical symmetry plane whose nominal voltage is 440 kV and line length is 500 km.

Sufentanil-2-hydroxypropyl-beta-cyclodextrin inclusion complex for pain treatment: Physicochemical, cytotoxicity, and pharmacological evaluation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaluation of uranium incorporation from contaminated areas using teeth as bioindicators - a case study

The Southwest region of the Bahia state in Brazil hosts the largest uranium reserve of the country (100 kton in uranium, only), plus the cities of Caetite, Lagoa Real and Igapora. In this work, aim was at the investigation of uranium burdens on residents of these cities by using teeth as bioindicators, as a contribution for possible radiation protection measures. Thus, a total of 41 human teeth were collected, plus 50 from an allegedly uranium free area (the control region). Concentrations of uranium in teeth from residents of 5- to 87-y old were determined by means of a high-resolution inductively coupled plasma mass spectrometer (ICP-MS). The highest uranium concentration in teeth was measured from samples belonging to residents of Caetite (median equal to 16 ppb). Assuming that the uranium concentrations in teeth and bones are similar within 10-20% (for children and young adults), it concluded that uranium body levels in residents of Caetite are at least one order of magnitude higher than the worldwide average. This finding led to conclude that daily ingestion of uranium, from food and water, is equally high.

Application of methylotrophic yeast Pichia pastoris in the field of food industry - A review

Since ancient times, the utilization of yeasts by the man has a great impact on the socio-economic development. After the advent of the technology of recombinant DNA, great advances have occurred due to the acquisition of strains of mutant yeasts in the field of applied research, and Saccharomyces cerevisiae has soon been outstanding as an interesting candidate for the expression of heterologous proteins of biotechnological interest. As the time goes by other alternative systems of expression have been shown because they have advantages over Saccharomyces cerevisiae. Among those new systems, Pichia pastoris is outstanding as methylotrophic yeast capable of growing in a culture medium containing methanol as the only source of carbon and energy. The induction of production of glycerol-3-phosphate dehydrogenase (GPD, NAD(+): oxido-redutase EC 1.1. 1.8) by Pichia pastoris was accomplished in the medium containing methanol. One of the most important key parameters in Pichia pastoris expression system is the methanol concentration. Bibliographic reviews on the Pichia pastoris production system have shown that the best culture conditions vary according to the strain used and/or kind of heterologous protein desired to be expressed. Therefore, we have sought to develop a system, involving expression of glycerol-3-phosphate dehydrogenase in the yeast Pichia pastoris, for generating sufficient quantities of the enzyme in order to asses its potential value for use in various food bioanalytical determination. Dehydrogenases have been widely used in the enzymatic assays of diverse composites of industrial interest, being enclosed among them glycerol and a number of important bioanalytical applications.

Evaluation of modifiers for determination of V in parsley by GF AAS

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In Vitro Evaluation of the Genotoxic Activity and Apoptosis Induction of the Extracts of Roots and Leaves from the Medicinal Plant Coccoloba mollis (Polygonaceae)

Coccoloba mollis (Family Polygonaceae) is a medicinal plant popularly used in cases of memory loss, stress, insomnia, anemia, impaired vision, and sexual impotence, but the scientific literature, to date, lacks studies on the biological effects of this species, particularly with regard to cytotoxicity and induction of DNA damage. The aim of the present study was to assess in vitro (in hepatic HTC cells) ethanolic extracts of the roots and leaves of C. mollis for cytotoxicity, genotoxicity, and induction of apoptosis. For these evaluations the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay, comet assay, micronucleus test with cytokinesis block, and an in situ test for detection of apoptotic cells with acridine orange staining were used. The results showed that the extract obtained from the roots of C. mollis is more cytotoxic than that obtained from the leaves and that the reduction in cell viability observed in the MTT assay was a result, at least in part, from the induction of apoptosis. Both extracts induced DNA damage at a concentration of 20 mu g/mL in the comet assay, but no genotoxicity was detected with any of the treatments carried out in the micronucleus test.

Isolation and evaluation of the antioxidant activity of phenolic constituents of the Garcinia brasiliensis epicarp

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Morfina nasal: Avaliacao da analgesia e dos efeitos colaterais no tratamento da dor pos-operatoria em criancas

Background and Objectives - Postoperative pain is one of the major discomforts but often under treated, especially in the pediatric patient. The aim of this study was to evaluate nasal morphine postoperative analgesia as an alternative drug administration route and show its applicability, effectiveness, tolerability and side effects. Methods - Participated in this study 20 patients aged 3 to 13 years, physical status ASA I and II sequentially submitted the different small and medium-size surgeries. Analgesia was obtained with nasal morphine hydrochloride in aqueous solution in variable concentrations of 2%, 1%, 0.5%, 0.25% and 0.125%. The dose for each instillation has been 0.1 mg.kg -1 at three-hour intervals for 36 postoperative hours. Quality of analgesia in pre-verbal age patients was evaluated by a pain intensity scale based on facial expression and crying, sleep, motor activity, sociability and food ingestion was used. Standardized evaluations were performed at 3-hour intervals. A four-grade scale was used to evaluate tolerability, where: 1) Good; 2) Regular; 3) Bad; 4) Very bad. Result - Postoperative analgesia results have proven to be good and safe, especially from the third evaluation on (6 hours). Drug tolerability has been good, although side effects were observed, especially nausea and vomiting. Conclusions - Patients and relatives accepted the method very well. The nasal route was considered an adequate way for opioid administration although more studies are needed to accept it as a routine for postoperative morphine analgesia.

Expanded food choice as a possible factor in the evolution of sociality of Vespidae (Hymenoptera)

A recent theory suggests that economic considerations are more important than genetic ones in the emergence and maintenance of social behavior. Evolution of social behavior in wasps, thus, could be based on the development of worker castes, which increase the efficiency of brood care and energy use of the colony. If so, social wasps should collect a larger range of prey, favoring polyethism, as social behavior should increase the adaptive value of social species among wasps by increasing the range of prey accessible. We explored the literature and showed that the Eumeninae, which are mostly solitary, draw prey from significantly fewer orders of arthropods than wasps in the subfamily Vespinae and Polistinae, which are mainly social, supporting the hypothesis that social behavior may have emerged as a more efficient way to feed and care for the young by opening a wider range of food sources, increasing the amount of food and quality of care provided to the young. Two alternative explanations of this data are also discussed.