Natural history of Peucetia flava (Araneae, Oxyopidae): seasonal density fluctuation, phenology and sex ratio on the glandular plant Rhyncanthera dichotoma (Melastomataceae)

The seasonal density fluctuation, phenology and sex ratio of Peucetia flava (Oxyopidae) on Rhyncanthera dichotoma (Melastomataceae) were investigated during a year in a swamp in southeastern Brazil. Peucetia flava displayed an unusual non-seasonal life cycle and the population size varied little over the year. The density of Peucetia spiders increased with the increase in abundance of leaves and number of arthropods adhered to glandular trichomes on R. dichotoma leaves. Our findings suggest that seasonal density fluctuation of Peucetia may be influenced by foraging site availability (i.e. leaves) and prey supply (i.e. arthropods adhering to glandular trichomes). The unusual seasonal stability of P. flava may be related to the type of habitat in which this spider occurs (swamp), because of the constant input of allochthonous resources from the water source, i.e. aquatic insects that migrate to the terrestrial environment.

Development of environmental and natural vulnerability maps for Brazilian coastal at Sao Sebastiao in São Paulo State

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

CNS-selective noncompetitive cholinesterase inhibitors derived from the natural piperidine alkaloid (-)-spectaline

LASSBio-767 [(-)-3-O-acetyl-spectaline] and LASSBio-822 [(-)-3-O-tert-Boc-spectaline] were recently described as cholinesterase inhibitors derived from the natural piperidine alkaloid (-)-spectaline, obtained from the flowers of Senna spectabilis (Fabaccae). We investigated their mechanism of inhibition of acetylcholinesterase and their efficacy in reversing scopolamine-induced amnesia. Competition assays with the substrate acetylthiocholine showed a concentration-dependent reduction in rat brain cholinesterase V-max without changes in apparent K-m. The kinetic data for LASSBio-767 and LASSBio-822 were best fit by a model of simple linear noncompetitive inhibition with K-i of 6.1 mu M and 7.5 mu M, respectively. A dilution assay showed a fast and complete reversal of inhibition, independent of incubation time. Simulated docking of the compounds into the catalytic gorge of Torpedo acetylcholinesterase showed interactions with the peripheral anionic site, but not with the catalytic triad. Anti-amnestic effects in mice were assessed in a step-down passive avoidance test and in the Morris water maze 30 min after injection of scopolamine (1 mg/kg i.p.). Saline, LASSBio-767, or LASSBio-822 was administered 15 min before scopolamine. Both compounds reversed the scopolamine-induced reduction in step-down latency at 0.1 mg/kg i.p. LASSBio-767 reversed scopolamine-induced changes in water maze escape latency at 1 mg/kg i.p. or p.o., while its cholinergic side effects were absent or mild up to 30 mg/kg i.p. (LD50 above 100 mg/kg i.p.). Thus, the (-)-spectaline derivatives are potent cholinergic agents in vivo, with a unique profile combining noncompetitive cholinesterase inhibition and CNS selectivity, with few peripheral side effects. (C) 2007 Elsevier B.V. All rights reserved.

Whydration effects on DNA double helix stability modulates ligand binding to natural DNA in response to changes in water activity

In this work we present evidence that water molecules are actively involved on the control of binding affinity and binding site discrimination of a drug to natural DNA. In a previous study, the effect of water activity (a(w)) on the energetic parameters of actinomycin-D intercalation to natural DNA was determined using the osmotic stress method (39). This earlier study has shown evidence that water molecules act as an allosteric regulator of ligand binding to DNA via the effect of water activity on the long-range stability of the DNA secondary structure. In this work we have carried out DNA circularization experiments using the plasmid pUC18 in the absence of drugs and in the presence of different neutral solutes to evaluate the contribution of water activity to the energetics of DNA helix unwinding. The contribution of water to these independent reactions were made explicit by the description of how the changes in the free energy of ligand binding to DNA and in the free energy associated with DNA helix torsional deformation are linked to a(w) via changes in structural hydration. Taken together, the results of these studies reveal an extensive linkage between ligand binding affinity and site binding discrimination, and long range helix conformational changes and DNA hydration, This is strong evidence that water molecules work as a classical allosteric regulator of ligand binding to the DNA via its contribution to the stability of the double helix secondary structure, suggesting a possible mechanism by which the biochemical machinery of DNA processing takes advantage of the low activity of water into the cellular milieu.

Active and exo-site inhibition of human factor Xa: Structure of des-Gla factor Xa inhibited by NAP5, a potent nematode anticoagulant protein from Ancylostoma caninum

Hookworms are hematophagous nematodes capable of growth, development and subsistence in living host systems such as humans and other mammals. Approximately one billion, or one in six, people worldwide are infected by hookworms causing gastrointestinal blood loss and iron deficiency anemia. The hematophagous hookworm Ancylostoma caninum produces a family of small, disulfide-linked protein anticoagulants (75-84 amino acid residues). One of these nematode anticoagulant proteins, NAP5, inhibits the amidolytic activity of factor Xa (fXa) with K-i = 43 pM, and is the most potent natural fXa inhibitor identified thus far. The crystal structure of NAP5 bound at the active site of gamma-carboxyglutamic acid domainless factor Xa (des-fXa) has been determined at 3.1 angstrom resolution, which indicates that Asp189 (fXa, S1 subsite) binds to Arg40 (NAP5, P1 site) in a mode similar to that of the BPTI/trypsin interaction. However, the hydroxyl group of Ser39 of NAP5 additionally forms a hydrogen bond (2.5 angstrom) with His57 NE2 of the catalytic triad, replacing the hydrogen bond of Ser195 OG to the latter in the native structure, resulting in an interaction that has not been observed before. Furthermore, the C-terminal extension of NAP5 surprisingly interacts with the fXa exosite of a symmetry-equivalent molecule forming a short intermolecular beta-strand as observed in the structure of the NAPc2/fXa complex. This indicates that NAP5 can bind to fXa at the active site, or the exosite, and to fX at the exosite. However, unlike NAPc2, NAP5 does not inhibit fVIIa of the fVIIa/TF complex. (c) 2007 Elsevier Ltd. All rights reserved.

Aerosol and precipitation chemistry measurements in a remote site in Central Amazonia: the role of biogenic contribution

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

Intercontinental Differences in the Abundance of Solenopsis Fire Ants (Hymenoptera: Formicidae): Escape from Natural Enemies?

The absence of natural enemies often allows exotic pests to reach densities that are much higher than normally occur in their native habitats. When Solenopsis fire ants were introduced into the United States, their numerous natural enemies were left behind in South America. To compare intercontinental fire ant densities, we selected 13 areas in South America and another 12 areas in North America. Sample areas were paired with weather stations and distributed across a broad range of climatic conditions. In each area, we measured fire ant densities at 5 preselected roadside sites that were at least 5 km apart. At each site, we also measured foraging activity, checked for polygyne colonies, and recorded various kinds of environmental data. In most areas, we also measured fire ant densities in lawns and grazing land. Fire ant populations along roadsides in North America were 4-7 times higher than fire ant populations in South America. Similar intercontinental differences were found in lawns and on grazing lands. These intercontinental differences in fire ant abundance were not associated with sampling conditions, seasonal variability, habitat differences, or the frequency of polygyny. Although several correlations were found with long-term weather conditions, careful inspection of the data suggests that these correlations were probably more coincidental than causal. Cultural differences in roadside maintenance may explain some of the intercontinental differences in fire ant abundance, but they did not account for equivalent intercontinental differences in grazing land and mowed lawns. Bait tests showed that competition with other ants was much more important in South America; however, we were not able to determine whether this was a major cause of intercontinental differences or largely a consequence of other factors such as the numerous pathogens and parasites that are found in South America. Because this study was correlational, we were unable to determine the cause(s) of the large intercontinental difference in fire ant abundance that we observed. However, we were able to largely exclude a number of possible explanations for the differences, including sampling, season, polygyny, climate, and aspects of habitat. By a process of elimination, escape from natural enemies remains among the most likely explanations for the unusually high densities of fire ants found in North America.

Few substitutions affect the bioluminescence spectra of Phrixotrix (Coleoptera: Phengodidae) luciferases: a site-directed mutagenesis survey

Phrixotrix (railroad worm) luciferases produce bioluminescence in the green and red regions of the spectrum, depending on the location of the lanterns, and are the only luciferases naturally producing red bioluminescence. Comparison of the luciferase sequences showed a set of substitutions that could be involved in bioluminescence colour determination: (a) unique substitutions in the red luciferase replacing otherwise invariant residues; (b) conserved basic residues in the green-yellow emitting luciferases; and (c) an additional R353 residue in red-emitting luciferase (Viviani et al., 1999). To investigate whether these sites have a functional role in bioluminescence colour determination, we performed a site-directed mutagenesis. Natural substitutions in the region 220-344 and residues in the putative luciferin-binding site were also investigated. With the exception of the previously identified substitution of R215 and T226 (Viviani et al., 2002), which display dramatic red-shift effects on the spectrum of green-yellow-emitting luciferases, only a few substitutions had a moderate effect on the spectrum of the green-emitting luciferase. In contrast, no single substitution affected the spectrum of the red-emitting luciferase. The results suggest that the identity of the active site residues is not so critical for determining red bioluminescence in PxRE luciferase. Rather, the conformation assumed during the emitting step could be critical to set up proper interactions with excited oxyluciferin. Copyright ©2007 John Wiley & Sons, Ltd.

Biochemical and hematological study of goats envenomed with natural and 60Co-irradiated bothropic venom

Venoms from snakes of the Bothrops genus are proteolytic, coagulant, hemorrhagic and nephrotoxic, causing edema, necrosis, hemorrhage and intense pain at the bite site, besides systemic alterations. Many adjuvants have been added to the venom used in the sensitization of antiserum-producer animals to increase antigenic induction and reduce the envenomation pathological effects. Gamma radiation from 60Co has been used as an attenuating agent of the venoms toxic properties. The main objective was to study, comparatively, clinical and laboratory aspects of goats inoculated with bothropic (Bothrops jararaca) venom, natural and irradiated from a 60Co source. Twelve goats were divided into two groups of six animals: GINV, inoculated with 0.5mg/kg of natural venom; and GIIV, inoculated with 0.5mg/kg of irradiated venom. Blood samples were collected immediately before and one, two, seven, and thirty days after venom injection. Local lesions were daily evaluated. The following exams were carried out: blood tests; biochemical tests of urea, creatinine, creatine kinase (CK), aspartate amino-transferase (AST) and alanine amino-transferase (ALT); clotting time; platelets count; and total serum immunoglobulin measurement. In the conditions of the present experiment, irradiated venom was less aggressive and more immunogenic than natural venom.

Astrocyte intermediate filaments are important markers in olfactory bulb of bovine herpesvirus type 5 natural infections

Astroglial cells are the most abundant cells in the mammalian central nervous system, yet our knowledge about their function in bovine Herpesvirus type 5 (BoHV-5) has been limited. The aim of this study was to detect by immunohistochemistry assay the reactive astrocytes for glial fibrilary acidic protein (GFAP) and vimentin (VIM), considered intermediate filaments of the cytoskeleton, localized in olfactory bulb from natural acute cases of BoHV-5 infection. All samples were submitted to virus isolation, real-time polymerase chain reaction (RT-PCR) and in situ hybridization (ISH) technique to confirm the virus transcription and respective genome. Samples were classified into four groups according to the severity of histological lesions. Groups III and IV, which histological lesions were classified as alacia, gliosis, satellitosis, neuronophagia and neuronal necrosis, 35% (± 1.8-2.1) of the inflammatory mononuclear cells, corresponded to CD3 positive lymphocytes. In the same group, 35% (± 1.8) of astrocytes were described as reactive to GFAP and VIM proteins. An agreement of r = 1.0 (P<0.0001) was found between histological lesions, intermediate filaments expression, viral DNA and transcription and CD3 lymphocytes. However, samples with mild histological lesions, 10.8 to 14.2% of astrocytes were classified as reactive to GFAP and VIM filaments. Our findings suggest that GFAP and VIM reactive astrocytes, in primary site of virus replication, seems to play an important role in neurovirulence, in spite of many questions concerning the virus immunopathology remains unclear.

Development of a natural products database from the biodiversity of Brazil

We describe herein the design and development of an innovative tool called the NuBBE database (NuBBEDB), a new Web-based database, which incorporates several classes of secondary metabolites and derivatives from the biodiversity of Brazil. This natural product database incorporates botanical, chemical, pharmacological, and toxicological compound information. The NuBBEDB provides specialized information to the worldwide scientific community and can serve as a useful tool for studies on the multidisciplinary interfaces related to chemistry and biology, including virtual screening, dereplication, metabolomics, and medicinal chemistry. The NuBBEDB site is at http://nubbe.iq.unesp.br/nubbeDB.html. © 2013 The American Chemical Society and American Society of Pharmacognosy.

Estudos limnológicos de balneabilidade no Parque Natural Municipal Cachoeira da Marta (Botucatu, SP, Brasil): relação com possíveis fontes poluidoras

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo comparativo entre peritônio bovino e biomembrana de látex natural na substituição de fragmento da parede da bexiga em coelhos (Oryctolagus cuniculus)

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

Levantamento da entomofauna em sítios florestais em recuperação e em um fragmento de floresta natural em Botucatu, SP

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Descrição taxonômica e história natural de uma nova espécie de Hypsiboas da Mata Atlântica do Alto da Serra de Paranapiacaba, Estado de São Paulo (Amphibia, Anura, Hylidae)

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