Comparative cytogenetic analysis of eleven species of subfamilies Neoplecostominae and Hypostominae (Siluriformes : Loricariidae)

The family Loricariidae with about 690 species divided into six subfamilies, is one of the world's largest fish families. Recent studies have shown the existence of several problems in the definition of natural groups in the family, which has made the characterization of the subfamilies and even of some genera quite difficult. With the main objective of contributing for a better understanding of the relationships between loricariids, cytogenetic analysis were conducted with two species of Neoplecostominae and nine species of Hypostominae that, according to morphological and molecular data, may belong to a new monophyletic unit. The results obtained showed a marked chromosomal conservation with the presence of 2n = 54 chromosomes and single interstitial Ag-NORs in all species analyzed. Considering that Neoplecostominae is the primitive sister-group of all other loricariids, with exception of Lithogeneinae, this karyotypic structure may represent the primitive condition for the family Loricariidae. The cytogenetic characteristics partaken by the species of Neoplecostominae and Hypostominae analyzed in the present study reinforce the hypothesis that the species of both these subfamilies might belong to a natural group.

Development of an operational substrate for ZapA, a metalloprotease secreted by the bacterium Proteus mirabilis

The protease ZapA, secreted by Proteus mirabilis, has been considered to be a virulence factor of this opportunistic bacterium. The control of its expression requires the use of an appropriate methodology, which until now has not been developed. The present study focused on the replacement of azocasein with fluorogenic substrates, and on the definition of enzyme specificity. Eight fluorogenic substrates were tested, and the peptide Abz-Ala-Phe-Arg-Ser-Ala-Ala-Gln-EDDnp was found to be the most convenient for use as an operational substrate for ZapA. A single peptide bond (Arg-Ser) was cleaved with a Km of 4.6 µM, a k cat of 1.73 s-1, and a catalytic efficiency of 376 (mM s)-1. Another good substrate for ZapA was peptide 6 (Abz-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-Gln-EDDnp) which was cleaved at a single bond (Phe-Ser) with a Km of 13.6 µM, a k cat of 3.96 s-1 and a catalytic efficiency of 291 (mM s)-1. The properties of the amino acids flanking the scissile bonds were also evaluated, and no clear requirement for the amino acid residue at P1 was found, although the enzyme seems to have a preference for a hydrophobic residue at P2.

The use of biodiversity as source of new chemical entities against defined molecular targets for treatment of malaria, tuberculosis, and T-cell mediated diseases: a review

The modern approach to the development of new chemical entities against complex diseases, especially the neglected endemic diseases such as tuberculosis and malaria, is based on the use of defined molecular targets. Among the advantages, this approach allows (i) the search and identification of lead compounds with defined molecular mechanisms against a defined target (e.g. enzymes from defined pathways), (ii) the analysis of a great number of compounds with a favorable cost/benefit ratio, (iii) the development even in the initial stages of compounds with selective toxicity (the fundamental principle of chemotherapy), (iv) the evaluation of plant extracts as well as of pure substances. The current use of such technology, unfortunately, is concentrated in developed countries, especially in the big pharma. This fact contributes in a significant way to hamper the development of innovative new compounds to treat neglected diseases. The large biodiversity within the territory of Brazil puts the country in a strategic position to develop the rational and sustained exploration of new metabolites of therapeutic value. The extension of the country covers a wide range of climates, soil types, and altitudes, providing a unique set of selective pressures for the adaptation of plant life in these scenarios. Chemical diversity is also driven by these forces, in an attempt to best fit the plant communities to the particular abiotic stresses, fauna, and microbes that co-exist with them. Certain areas of vegetation (Amazonian Forest, Atlantic Forest, Araucaria Forest, Cerrado-Brazilian Savanna, and Caatinga) are rich in species and types of environments to be used to search for natural compounds active against tuberculosis, malaria, and chronic-degenerative diseases. The present review describes some strategies to search for natural compounds, whose choice can be based on ethnobotanical and chemotaxonomical studies, and screen for their ability to bind to immobilized drug targets and to inhibit their activities. Molecular cloning, gene knockout, protein expression and purification, N-terminal sequencing, and mass spectrometry are the methods of choice to provide homogeneous drug targets for immobilization by optimized chemical reactions. Plant extract preparations, fractionation of promising plant extracts, propagation protocols and definition of in planta studies to maximize product yield of plant species producing active compounds have to be performed to provide a continuing supply of bioactive materials. Chemical characterization of natural compounds, determination of mode of action by kinetics and other spectroscopic methods (MS, X-ray, NMR), as well as in vitro and in vivo biological assays, chemical derivatization, and structure-activity relationships have to be carried out to provide a thorough knowledge on which to base the search for natural compounds or their derivatives with biological activity.

Toward a hierarchical concept of plant stress

A number of attempts have been made to obtain a clear definition of biological stress. However, in spite of the efforts, some controversies on the concept of plant stress remain. The current versions are centered either on the cause (stress factor) or on the effect (stress response) of environmental stress. The objective of this study was to contribute to the definition of stress, using a hierarchical approach. Thus, we have performed an analysis of the most usual stress concepts and tested the relevance of considering different observation scales in a study on plant response to water deficit. Seedlings of Eucalyptus grandis were grown in vitro at water potentials ranging from -0.16 to -0.6 MPa, and evaluated according to growth and biochemical parameters. Data were analyzed through principal component analysis (PCA), which pointed to a hierarchical organization in plant responses to environmental disturbances. Growth parameters (height and dry weight) are more sensitive to water deficit than biochemical ones (sugars, proline, and protein), suggesting that higher hierarchical levels were more sensitive to environmental constraints than lower hierarchical ones. We suggest that before considering an environmental fluctuation as stressful, it is necessary to take into account different levels of plant response, and that the evaluation of the effects of environmental disturbances on an organism depends on the observation scale being used. Hence, a more appropriate stress concept should consider the hierarchical organization of the biological systems, not only for a more adequate theoretical approach, but also for the improvement of practical studies on plants under stress.

Gap size measurement: the proposal of a new field method

In the tropical Atlantic Forest, 42 canopy gaps had their areas estimated using four different field methods of measurement: Runkle, Brokaw and Green [Runkle, J.R., 1981. Gap formation in some old-growth forests of the eastern United States. Ecology 62, 1041-1051; Brokaw, N.V.L., 1982. The definition of treefall gap and its effect on measures of forest dynamics. Biotropica 14, 158-160; Green, P.T., 1996. Canopy Gaps in rain forest on Christmas Island, Indian Ocean: size distribution and methods of measurement. J. Trop. Ecol. 12, 427-434] and a new method proposed in this work. It was found that within the same gap delimitation, average gap size varied from 56.0 up to 88.3 m(3) while total sum of gap area varied from 2351.3 to 3707.9 m(3) Differences among all methods and between pairs of method proved to be statistically significant. As a consequence, gap size-class distribution was also different between methods. When one method is held as a standard, deviation on average values of gap size ranged between 11.8 and 59.7% as deviations on single gap size can reach 172.8%. Implications on forest dynamics were expressed by the forest turnover rate that was 24% faster or 15% slower depending on the method adopted for gap measurement. Based on my results and on methods' evaluation, the use of a new method is proposed here for future research involving the measure of gap size in forest ecosystems. Finally, it is concluded that forest comparisons disregarding the influence of different methods of gap measurement should be reconsidered. (c) 2005 Elsevier B.V. All rights reserved.

Fish as Indicators of Disturbance in Streams Used for Snorkeling Activities in a Tourist Region

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Evaluation of Staining Methods for Cytologic Diagnosis of Oral Lesions

ObjectiveTo compare the efficacy of Papanicolaou, hematoxylin-eosin (H-E), Leishman and periodic acid-Schiff (PAS) staining for Cytologic diagnosis of oral lesions.Study DesignPatients from the Discipline of Stomatology, Sao Jose dos Campos Dental School, from the wards of Hospital Heliopolis and from the dentistry outpatient clinic of the University Hospital, University of São Paulo Medical School, with the following diseases, were selected: erythematous candidiasis (n = 9), pseudomembranous candidiasis (n = 10), squamous cell carcinoma In = 19), herpes simplex (n = 8), paracoccidioidomycosis In = 8) and pemphigus vulgaris (n = 1).ResultsThe different staining methods were compared regarding the quality of definition of cytoplasmic and nuclear morphologic characteristics and the identification of bacteria, fungi, inflammatory cells and secretions. Papanicolaou and H-E staining were considered better methods. In cases of fungal infections, PAS staining is useful and should be applied as a complementary method.ConclusionWithin the limitations of this study, it can be concluded that the cytologic diagnosis of oral lesions along with different staining methods is a useful tool for oral diagnosis. (Acta Cytol 2008;52:697-701)

Orthogonal projections and bootstrap resampling procedures in the study of infraspecific variation

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

NORDSIECK FORMS OF STIFFLY-STABLE ADAMS-TYPE CYCLIC METHODS THAT ARE INSENSITIVE TO STEP-SIZE CHANGES

In this paper we determine a matrix S and a vector l for stiffly-stable Adams-type cyclic methods that are insensitive to step size changes by using the definition of equivalent methods, (see, e.g. [l]), in the Nordsieck notation. The elements S and l, written in a parametric form, permit us to represent in Nordsieck form the methods that were constructed in [7] and the new methods that satisfy the above properties.

Some particularities of dam rehabilitation projects

The number of dams, which need rehabilitation, is growing, not only in countries that have a long tradition in dam building and operation but,also in those regions where the infrastructure is still in full development. Though rehabilitation projects generally deal with problems that are common in dam engineering practice there are some peculiarities which are a characteristic of such projects and which must be duly taken into account to avoid unsuccess and/or unnecessary costs. Regular safety inspection is essential to forestall the development of structural, hydrological and operational unsafety. if need of major repair or overall rehabilitation of a dam becomes apparent design oft he rehabilitation project must be preceded by a comprehensive checkup of the structure and appurtenant works, as well as by an evaluation of its hydrological safety inclusive of all relevant environmental aspects. The availability of complete records on the clam's structural behaviour and on meteorological and hydrological data, as well as the knowledge of the materials properties of the existing structure are important for the successful design of a rehabilitation project. To this end the installation of monitoring devices in the existing structure may be necessary to generate representative data. While the criteria to be used in structural design should correspond to current standards, the definition of hydrological design criteria depends on considerations that vary widely from region to region or even from one country to another. Some basic hydrological safety requirements, however, are recommended for general acceptance. Dam rehabilitation projects demand very careful and detailed construction planning because of their dependence on river flow conditions, operational restrictions and, often, on procedures or limitations imposed to avoid harm to the environment. of utmost importance is the timely availability of the financial funds required to complete the project, in order to avoid delays which could result in structural or operational unsafety. Since every dam sooner or later will have to undergo major repair or updating of safety, rehabilitation may evolve to a speciality of dam engineering.

Serotonergic mechanisms of the lateral parabrachial nucleus and cholinergic-induced sodium appetite

Central cholinergic mechanisms are suggested to participate in osmoreceptor-induced water intake. Therefore, central injections of the cholinergic agonist carbachol usually produce water intake (i.e., thirst) and are ineffective in inducing the intake of hypertonic saline solutions (i.e., the operational definition of sodium appetite). Recent studies have indicated that bilateral injections of the serotonin receptor antagonist methysergide into the lateral parabrachial nucleus (LPBN) markedly increases salt intake in models involving the activation of the renin-angiotensin system or mineralocorticoid hormones. The present studies investigated whether sodium appetite could be induced by central cholinergic activation with carbachol (an experimental condition where only water is typically ingested) after the blockade of LPBN serotonergic mechanisms with methysergide treatment in rats. When administered intracerebroventricularly in combination with injections of vehicle into both LPBN, carbachol (4 nmol) caused water drinking but insignificant intake of hypertonic saline. In contrast, after bilateral LPBN injections of methysergide (4 mug), intracerebroventricular carbachol induced the intake of 0.3 M NaCl. Water intake stimulated by intracerebroventricular carbachol was not changed by LPBN methysergide injections. The results indicate that central cholinergic activation can induce marked intake of hypertonic NaCl if the inhibitory serotonergic mechanisms of the LPBN are attenuated.

Regularization of discontinuous vector fields on R-3 via singular perturbation

Singular perturbations problems in dimension three which are approximations of discontinuous vector fields are studied in this paper. The main result states that the regularization process developed by Sotomayor and Teixeira produces a singular problem for which the discontinuous set is a center manifold. Moreover, the definition of' sliding vector field coincides with the reduced problem of the corresponding singular problem for a class of vector fields.

Insights into the concept of fish welfare

Fish welfare issues are predicated on understanding whether fish are sentient beings. Therefore, we analyzed the logic of the methodologies used for studying this attribute. We conclude that empirical science is unable to prove or to disprove that fish are sentient beings. Thus, we propose a combined ethical-scientific approach for considering fish as sentient beings. The most difficult ongoing question is to determine which conditions fish prefer. Approaches to assess fish preferences should be rigorously and cautiously employed. In light of these considerations, attempts to establish physiological standards for fish welfare are discouraged, and a preference-based definition of fish welfare is proposed.

COMPARATIVE INVITRO ACTIVITY OF 3 CEPHALOSPORINS AGAINST STAPHYLOCOCCUS-AUREUS ISOLATED FROM BOVINE MASTITIS

This report compares the in vitro activity of three cephalosporins (cephalothin, cefoxitin and ceftriaxone) against 57 Staphylococcus aureus strains isolated from cows with clinical mastitis on the basis of the minimal inhibitory (MIC) and minimal bactericidal concentrations (MBC). The majority of the S aureus strains showed resistance to cefoxitin and ceftriaxone and sensitivity to cephalothin. The highest MICs and MBCs were found for cefoxitin and ceftriaxone. Antimicrobial tolerance (MBC/MIC greater-than-or-equal-to 32:1) was observed in relation to cephalothin and ceftriaxone. The data suggest that these cephalosporins may not be effective for the treatment of staphylococcal bovine mastitis. The precise definition of their antimicrobial efficacies requires more detailed in vitro and in vivo studies.

The generation and utilization of a cancer-oriented representation of the human transcriptome by using expressed sequence tags

Whereas genome sequencing defines the genetic potential of an organism, transcript sequencing defines the utilization of this potential and links the genome with most areas of biology. To exploit the information within the human genome in the fight against cancer, we have deposited some two million expressed sequence tags (ESTs) from human tumors and their corresponding normal tissues in the public databases. The data currently define approximate to23,500 genes, of which only approximate to1,250 are still represented only by ESTs. Examination of the EST coverage of known cancer-related (CR) genes reveals that <1% do not have corresponding ESTs, indicating that the representation of genes associated with commonly studied tumors is high. The careful recording of the origin of all ESTs we have produced has enabled detailed definition of where the genes they represent are expressed in the human body. More than 100,000 ESTs are available for seven tissues, indicating a surprising variability of gene usage that has led to the discovery of a significant number of genes with restricted expression, and that may thus be therapeutically useful. The ESTs also reveal novel nonsynonymous germline variants (although the one-pass nature of the data necessitates careful validation) and many alternatively spliced transcripts. Although widely exploited by the scientific community, vindicating our totally open source policy, the EST data generated still provide extensive information that remains to be systematically explored, and that may further facilitate progress toward both the understanding and treatment of human cancers.