Production, purification, and characterization of an extracellular acid protease from the marine Antarctic yeast Rhodotorula mucilaginosa L7

The production, purification, and characterization of an extracellular protease released by Rhodotorula mucilaginosa L7 were evaluated in this study. This strain was isolated from an Antarctic marine alga and previously selected among others based on the capacity to produce the highest extracellular proteolytic activity in preliminary tests. R. mucilaginosa L7 was grown in Saboraud-dextrose medium at 25 °C, and the cell growth, pH of the medium, extracellular protease production and the glucose and protein consumption were determined as a function of time. The protease was then purified, and the effects of pH, temperature, and salt concentration on the catalytic activity and enzyme stability were determined. Enzyme production started at the beginning of the exponential phase of growth and reached a maximum after 48 h, which was accompanied by a decrease in the pH as well as reductions of the protein and glucose concentrations in the medium. The purified protease presented optimal catalytic activity at pH 5.0 and 50 °C. Finally, the enzyme was stable in the presence of high concentrations of NaCl. These characteristics are of interest for future studies and may lead to potential biotechnological applications that require enzyme activity and stability under acidic conditions and/or high salt concentrations.

Chemical and rheological properties of exopolysaccharides produced by four isolates of rhizobia

The rheological, physicochemical properties, emulsification and stability of exopolysaccharides (EPSs) from four rhizobia isolates (LBMP-C01, LBMP-C02, LBMP-C03 and LBMP-C04) were studied. The EPS yields of isolates under these experimental conditions were in the range of 1.5-6.63gL(-1). The LBMP-C04 isolate, which presented the highest EPS production (6.63gL(-1)), was isolated from Arachis pintoi and was identified as a Rhizobium sp. strain that could be explored as a possible potential source for the production of extracellular heteropolysaccharides. All polymers showed a pseudoplastic non-Newtonian fluid behavior or shear thinning property in aqueous solutions. Among the four EPS tested against hydrocarbons, EPS LBMP-C01 was found to be more effective against hexane, olive and soybean oils (89.94%, 82.75% and 81.15%, respectively). Importantly, we found that changes in pH (2-11) and salinity (0-30%) influenced the emulsification of diesel oil by the EPSs. EPSLBMP-C04 presented optimal emulsification capacity at pH 10 (E24=53%) and 30% salinity (E24=27%). These findings contribute to the understanding of the influence of the chemical composition, physical properties and biotechnology applications of rhizobial EPS solutions their bioemulsifying properties.

Straight and angulated abutments in platform switching: influence of loading on bone stress by three-dimensional finite element analysis

PURPOSE: In view of reports in the literature on the benefits achieved with the use of platform switching, described as the use of an implant with a larger diameter than the abutment diameter, the goal being to prevent the (previously) normal bone loss down to the first thread that occurs around most implants, thus enhancing soft tissue aesthetics and stability and the need for implant inclination due to bone anatomy in some cases, the aim of this study was to evaluate bone stress distribution on peri-implant bone, by using three-dimensional finite element analysis to simulate the influence of implants with different abutment angulations (0 and 15 degrees) in platform switching. METHODS: Four mathematical models of an implant-supported central incisor were created with varying abutment angulations: straight abutment (S1 and S2) and angulated abutment at 15 degrees (A1 and A2), submitted to 2 loading conditions (100 N): S1 and A1-oblique loading (45 degrees) and S2 and A2-axial loading, parallel to the long axis of the implant. Maximum (σmax) and minimum (σmin) principal stress values were obtained for cortical and trabecular bone. RESULTS: Models S1 and A1 showed higher σmax in cortical and trabecular bone when compared with S2 and A2. The highest σmax values (in MPa) in the cortical bone were found in S1 (28.5), followed by A1 (25.7), S2 (11.6), and A2 (5.15). For the trabecular bone, the highest σmax values were found in S1 (7.53), followed by A1 (2.87), S2 (2.85), and A2 (1.47). CONCLUSIONS: Implants with straight abutments generated the highest stress values in bone. In addition, this effect was potentiated when the load was applied obliquely.

Relation between bruxism and dental implants

OBJECTIVE: The aim of this study was to gather information and discuss the predictability of implant-supported prostheses in patients with bruxism by performing a literature review. METHODS: In order to select the studies included in this review, a detailed search was performed in PubMed and Medline databases, using the following key words: bruxism, dental implants, implant supported prosthesis, and dental restoration failure. Items that were included are: case reports, randomized controlled trials, in vitro studies, literature and systematic reviews, with or without meta-analysis, of the last 20 years that addressed the theme. Articles without abstracts, animal studies, articles in languages other than English and articles from journals unrelated to the dental field were excluded. RESULTS: after analysis according to inclusion and exclusion criteria, 28 articles were selected from a total of 54. It is known from the array of scientific articles which have assessed, either through retrospective, prospective or experimental studies, that the biomechanical and biological impact of bruxism on implant-supported prostheses is small, and that the literature has contributed little to exemplify the prosthetic limits of safety for the specialist from a clinical point of view. CONCLUSION: Although there is still no general consensus on this matter, most of the literature review articles do provide clinical guidelines that contribute to implant supported prostheses longevity and stability in patients with bruxism.

Validation of a stability-indicating HPLC method with diode array detection for the determination of beclomethasone dipropionate in aqueous suspension for nebulizer

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Caracterização de Híbridos Simples Experimentais de Milho (Zea Mays L.)

Pós-graduação em Agronomia - FEIS

HOME RANGES AND MOVEMENTS OF COYOTES IN THE NORTHERN CHIHUAHUAN DESERT

The coyote (Canis latrans) is among the most studied animals in North America. Because of its adaptability and success as a predator, the coyote has flourished and is still expanding its range. Coyotes can now be found throughout most of North America and south into Central America (Voight and Berg 1987). Studies in recent years have been extensive to understand the interrelationships of prey and coyotes (Shelton and Klindt 1974, Beckoff and Wells 1981), as well as demographic relationships (Davis et al. 1975, Knowlton and Stoddart 1978, Mitchell 1979, Bowen 1981) and feeding strategies (Todd and Keith 1976, Andelt et al. 1987, MacCracken and Hansen 1987, Gese et al. 1988a). With the advance of radio telemetry, researchers have investigated lifestyle characteristics spatially with home ranges or temporally with movements in relation to habitat requirements. Researchers have studied home ranges of coyotes in various regions of the United States (Livaitis and Shaw 1980, Andelt 1981, Springer 1982, Pyrah 1984, Gese et al. 1988a) and Canada (Bowen 1982). Some studies of home range were separated by season (Ozoga and Harger 1966) or relation to nearby food sources (Danner and Smith 1980). Home range analysis in relation to social interactions of coyotes has been either neglected, overlooked, or avoided. Gese et al. (1988a) recognized a transient class of coyote by home range size. Coyote social systems are very complex and can vary by season or locality in addition to some reports of group or pack systems (Hamlin and Schweitzer 1979, Beckoff and Wells 1981, Bowen 1981, Gese et al. 1988b). Coyotes maintain communication with conspecifics through vocal and olfactory signals (Lehner 1987, Bowen and McTaggert Cowan 1980). Social interactions may be by far the most complex and least understood aspect related to coyote ecology. Coyote movements can be related to many factors including food, water, cover, and social interactions. Movements in relation to food sources are well documented (Fitch 1948, Todd and Keith 1976, Danner and Smith 1980) although reports on movements in relation to water have not been reported, probably because of limited research in desert situations. There has been some mention of coyotes' movements in relation to cover (Wells and Beckoff 1982). The objectives of this study were to delineate annual and seasonal home ranges, movements, and habitat use of coyotes in the northern Chihuahuan desert.

Inhibitory activity of liposomal flavonoids during oxidative metabolism of human neutrophils upon stimulation with immune complexes and phorbol ester

Context and objective: The massive production of reactive oxygen species by neutrophils during inflammation may cause damage to tissues. Flavonoids act as antioxidants and have anti-inflammatory effects. In this study, liposomes loaded with these compounds were evaluated as potential antioxidant carriers, in attempt to overcome their poor solubility and stability. Materials and methods: Liposomes containing quercetin, myricetin, kaempferol or galangin were prepared by the ethanol injection method and analyzed as inhibitors of immune complex (IC) and phorbol ester-stimulated neutrophil oxidative metabolism by luminol (CLlum) and lucigenin-enhanced (CLluc) chemiluminescence (CL) assays. The mechanisms involved this activity of liposomal flavonoids, such as cytotoxicity and superoxide anion scavenging capacity, and their effect on phagocytosis of ICs were also investigated. Results and discussion: The results showed that the inhibitory effect of liposomal flavonoids on CLlum and CLluc is inversely related to the number of hydroxyl groups in the flavonoid B ring. Moreover, phagocytosis of liposomes by neutrophils does not seem to necessarily promote such activity, as the liposomal flavonoids are also able to reduce CL when the cells are pretreated with cytochalasin B. Under assessed conditions, the antioxidant liposomes are not toxic to the human neutrophils and do not interfere with IC-induced phagocytosis. Conclusion: The studied liposomes can be suitable carriers of flavonoids and be an alternative for the treatment of diseases in which a massive oxidative metabolism of neutrophils is involved.

Flexible and conservative features of social systems in tufted capuchin monkeys: comparing the socioecology of Sapajus libidinosus and Sapajus nigritus

Socioecological models assume that primates adapt their social behavior to ecological conditions, and predict that food availability and distribution, predation risk and risk of infanticide by males affect patterns of social organization, social structure and mating system of primates. However, adaptability and variation of social behavior may be constrained by conservative adaptations and by phylogenetic inertia. The comparative study of closely related species can help to identify the relative contribution of ecological and of genetic determinants to primate social systems. We compared ecological features and social behavior of two species of the genus Sapajus, S. nigritus in Carlos Botelho State Park, an area of Atlantic Forest in Sao Paulo state, and S. libidinosus in Fazenda Boa Vista, a semi-arid habitat in Piaui state, Brazil. S. libidinosus perceived higher predation risk and fed on clumped, high quality, and usurpable resources (fruits) all year round, whereas S. nigritus perceived lower predation risk and relied on evenly distributed, low-quality food sources (leaves) during periods of fruit shortage. As predicted by socioecology models, S. libidinosus females were philopatric and established linear and stable dominance hierarchies, coalitions, and grooming relationships. S. nigritus females competed less often, and could transfer between groups, which might explain the lack of coalitions and grooming bonds among them. Both populations presented similar group size and composition and the same polygynous mating system. The species differed from each other in accordance with differences in the characteristics of their main food sources, as predicted by socioecological models, suggesting that phylogenetic inertia does not constrain social relationships established among female Sapajus. The similarity in mating systems indicates that this element of the social system is not affected by ecological variables and thus, is a more conservative behavioral feature of the genus Sapajus. Am. J. Primatol. 74:315331, 2012. (c) 2011 Wiley Periodicals, Inc.

Dissecting structure-function-stability relationships of a thermostable GH5-CBM3 cellulase from Bacillus subtilis 168

Cellulases participate in a number of biological events, such as plant cell wall remodelling, nematode parasitism and microbial carbon uptake. Their ability to depolymerize crystalline cellulose is of great biotechnological interest for environmentally compatible production of fuels from lignocellulosic biomass. However, industrial use of cellulases is somewhat limited by both their low catalytic efficiency and stability. In the present study, we conducted a detailed functional and structural characterization of the thermostable BsCe15A (Bacillus subtilis cellulase 5A), which consists of a GH5 (glycoside hydrolase 5) catalytic domain fused to a CBM3 (family 3 carbohydrate-binding module). NMR structural analysis revealed that the Bacillus CBM3 represents a new subfamily, which lacks the classical calcium-binding motif, and variations in NMR frequencies in the presence of cellopentaose showed the importance of polar residues in the carbohydrate interaction. Together with the catalytic domain, the CBM3 forms a large planar surface for cellulose recognition, which conducts the substrate in a proper conformation to the active site and increases enzymatic efficiency. Notably, the manganese ion was demonstrated to have a hyper-stabilizing effect on BsCel5A, and by using deletion constructs and X-ray crystallography we determined that this effect maps to a negatively charged motif located at the opposite face of the catalytic site.

Three-dimensional solitons in cross-combined linear and nonlinear optical lattices

The existence and stability of three-dimensional (3D) solitons, in cross-combined linear and nonlinear optical lattices, are investigated. In particular, with a starting optical lattice (OL) configuration such that it is linear in the x-direction and nonlinear in the y-direction, we consider the z-direction either unconstrained (quasi-2D OL case) or with another linear OL (full 3D case). We perform this study both analytically and numerically: analytically by a variational approach based on a Gaussian ansatz for the soliton wavefunction and numerically by relaxation methods and direct integrations of the corresponding Gross-Pitaevskii equation. We conclude that, while 3D solitons in the quasi-2D OL case are always unstable, the addition of another linear OL in the z-direction allows us to stabilize 3D solitons both for attractive and repulsive mean interactions. From our results, we suggest the possible use of spatial modulations of the nonlinearity in one of the directions as a tool for the management of stable 3D solitons.

Viscosity of egg white from hens of different strains fed with commercial and natural additives

Yolk color and egg white (albumen) cleanliness and viscosity are important parameters by which consumers judge the quality of eggs. This study aimed to investigate changes in albumen viscosity during storage of eggs for up to 36 days from two different commercial laying hen strains (Carijo Barbada and Isa Brown) fed a diet containing annatto (1.5 and 2.0%) or a synthetic additive without synthetic colorants (control). Analyses of humidity, albumen height, pH, viscosity, foam formation, and stability were carried out on eggs. Carijo Barbada strain had smaller albumen, lower humidity and higher egg white viscosity than Isa Brown strain; however, with storage, viscosity lowered significantly on both strains. Initially, the addition of 2.0% of annatto or a synthetic additive increased viscosity in both strains, but with storage only the control maintained longer viscosity. Lower viscosity did not change foam density and stability.

Differences in diversity, structure, and variability between intertidal and subtidal meiofaunal assemblages

[EN] Meiofaunal assemblages from intertidal and shallow subtidal seabeds were studied at two sites (one dominated by volcanic sands and the other by organogenic sands) at Tenerife (Canary Islands, NE Atlantic Ocean) throughout an entire year (May 2000?April 2001). Specifically, we aimed (i) to test for differences in diversity, structure, and stability between intertidal and subtidal meiofaunal assemblages, and (ii) to determine if differences in the meiofaunal assemblage structure may be explained by environmental factors (granulometric composition, availability of organic matter, and carbonate content in sediments). A total of 103,763 meiofaunal individuals were collected, including 203 species from 19 taxonomic groups (Acari, Amphipoda, Cnidaria, Copepoda, Echinodermata, Gastrotricha, Isopoda, Insecta, Kinorrhyncha, Misidacea, Nematoda, Nemertini, Oligochaeta, Ostracoda, Polychaeta, Priapulida, Sipuncula, Tanaidacea, and Turbellaria). Nematodes were the most abundant taxonomic group. Species diversity was higher in the subtidal than in the intertidal zone at both sites, as a result of the larger dominance of a few species in the intertidal zone. The meiofaunal assemblage structure was different between tidal levels at both sites, the intertidal presenting greater temporal variability (multivariate dispersion) in the meiofaunal assemblage structure than the subtidal. Sediment grain size, here quantified by the different granulometric fractions, explained the variability in meiofaunal assemblage structure to a greater extent than the percentage of carbonates, a variable linked to sediment origin. This study revealed differences in diversity, assemblage structure, and variability between intertidal and subtidal meiofauna.

Supramolecular hybrid organic-inorganic multicomponent architectures in solution and on surface

Supramolecular architectures can be built-up from a single molecular component (building block) to obtain a complex of organic or inorganic interactions creating a new emergent condensed phase of matter, such as gels, liquid crystals and solid crystal. Further the generation of multicomponent supramolecular hybrid architecture, a mix of organic and inorganic components, increases the complexity of the condensed aggregate with functional properties useful for important areas of research, like material science, medicine and nanotechnology. One may design a molecule storing a recognition pattern and programming a informed self-organization process enables to grow-up into a hierarchical architecture. From a molecular level to a supramolecular level, in a bottom-up fashion, it is possible to create a new emergent structure-function, where the system, as a whole, is open to its own environment to exchange energy, matter and information. “The emergent property of the whole assembly is superior to the sum of a singles parts”. In this thesis I present new architectures and functional materials built through the selfassembly of guanosine, in the absence or in the presence of a cation, in solution and on the surface. By appropriate manipulation of intermolecular non-covalent interactions the spatial (structural) and temporal (dynamic) features of these supramolecular architectures are controlled. Guanosine G7 (5',3'-di-decanoil-deoxi-guanosine) is able to interconvert reversibly between a supramolecular polymer and a discrete octameric species by dynamic cation binding and release. Guanosine G16 (2',3'-O-Isopropylidene-5'-O-decylguanosine) shows selectivity binding from a mix of different cation's nature. Remarkably, reversibility, selectivity, adaptability and serendipity are mutual features to appreciate the creativity of a molecular self-organization complex system into a multilevelscale hierarchical growth. The creativity - in general sense, the creation of a new thing, a new thinking, a new functionality or a new structure - emerges from a contamination process of different disciplines such as biology, chemistry, physics, architecture, design, philosophy and science of complexity.

Researches on berry composition in red grape: agronomical, biochemical and molecular approaches

It is well known that the best grape quality can occur only through the achievement of optimal source/sink ratio. Vine balance is in fact a key parameter in controlling berry sugar, acidity and secondary metabolites content (Howell, 2001; Vanden Heuvel et al., 2004). Despite yield reduction and quality improvement are not always strictly related, cluster thinning is considered a technique which could lead to improvement in grape sugar and anthocyanin composition (Dokoozlian and Hirschfelt, 1995; Guidoni et al., 2002). Among several microclimatic variables which may impact grape composition, the effect of cluster light exposure and temperature, which probably act in synergistic and complex way, has been widely explored showing positive even sometimes contradictory results (Spayd et al., 2001; Tarara et al., 2008). Pre-bloom and véraison defoliation are very efficient techniques in inducing cluster microclimatic modification. Furthermore pre-bloom defoliation inducing a lower berry set percentage On these basis the aim of the first experiment of the thesis was to verify in cv Sangiovese the effects on ripening and berry composition of management techniques which may increase source/sink ratio and /or promote light incidence on berries throughout grape ripening. An integrated agronomic, biochemical and microarray approach, aims to understand which mechanisms are involved in berry composition and may be conditioned in the berries during ripening in vines submitted to three treatments. In particular the treatments compared were: a) cluster thinning (increasing in source/sink ratio) b) leaf removal at véraison (increasing cluster light exposure) c) pre-bloom defoliation (increasing source sink ratio and cluster light exposure). Vine response to leaf removal at véraison was further evaluated in the second experiment on three different varieties (Cabernet Sauvignon, Nero d’Avola, Raboso Piave) chosen for their different genetic traits in terms of anthocyanin amount and composition. The integrated agronomic, biochemical and microarray approach, employed in order to understand those mechanisms involved in berry composition of Sangiovese vines submitted to management techniques which may increase source/sink ratio and induce microclimatic changes, bring to interesting results. This research confirmed the main role of source/sink ratio in conditioning sugars metabolism and revealed also that carbohydrates availability is a crucial issue in triggering anthocyanin biosynthesis. More complex is the situation of pre-bloom defoliation, where source/sink and cluster light increase effects are associated to determine final berry composition. It results that the application of pre-bloom defoliation may be risky, as too much dependent on seasonal conditions (rain and temperature) and physiological vine response (leaf area recovery, photosynthetic compensation, laterals regrowth). Early induced stress conditions could bring cluster at véraison in disadvantage to trigger optimal berry ripening processes compared to untreated vines. This conditions could be maintained until harvest, if no previously described physiological recovery occurs. Certainly, light exposure increase linked to defoliation treatments, showed a positive and solid effect on flavonol biosynthesis, as in our conditions temperature was not so different among treatments. Except the last aspects, that could be confirmed also for véraison defoliation, microclimatic changes by themselves seemed not able to induce any modification in berry composition. Further studies are necessary to understand if the peculiar anthocyanic and flavonols composition detected in véraison defoliation could play important role in both color intensity and stability of wines.