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The objective of this study was to evaluate the effects of the addition of parts of leguminous trees on the growth and nutrition of maize (Zea mays L.), as well as on the chemical attributes of a degraded soil, 65 days after applying the residues. The experiment was conducted in pots, in a randomized block design with ten treatments and four replicates. The evaluated treatments were: T1 - No residues of leguminous trees, T2 - Leaves of Mimosa caesalpiniifolia, T3 - Branches of Mimosa caesalpiniifolia, T4 - Leaves + branches of Mimosa caesalpiniifolia, T5 - Leaves of Mimosa hostilis, T6 - Branches of Mimosa hostilis, T7 - Leaves + branches of Mimosa hostilis, T8 - Leaves of Gliricidia sepium, T9 - Branches of Gliricidia sepium and T10 - Leaves + branches of Gliricidia sepium. Pots were filled with soil from a degraded area and residues were added in the form of green mass after sowing the maize. Residues of leguminous trees positively influenced maize growth at 65 days after application and favored the accumulation of nitrogen, phosphorus and potassium in the shoots. Green fertilizers improved soil chemical attributes at 65 days after application, especially nitrate-N (NO3 --N), ammonium-N (NH4 +-N), total inorganic N (NO3 --N + NH4 +-N) and K, demonstrating that these species are good options for recovering degraded areas in the semi-arid region of Ceará. RESUMO Este estudo teve como objetivo avaliar o efeito da adição de resíduos de partes de leguminosas arbóreas no crescimento e nutrição de plantas de milho (Zea mays L.), bem como nos atributos químicos de um solo degradado, 65 dias após a aplicação dos resíduos. O experimento foi realizado em vasos, em delineamento de blocos ao acaso, com dez tratamentos e quatro repetições. Os tratamentos avaliados foram: T1 - Sem resíduo de leguminosas, T2 - Folhas de Mimosa caesalpiniifolia, T3 - Galhos de Mimosa caesalpiniifolia, T4 - Folhas + galhos de Mimosa caesalpiniifolia, T5 - Folhas de Mimosa hostilis, T6 - Galhos de Mimosa hostilis, T7 - Folhas + galhos de Mimosa hostilis, T8 - Folhas de Gliricidia sepium, T9 - Galhos de Gliricidia sepium e T10 - Folhas + galhos de Gliricidia sepium. Os vasos foram preenchidos com solo de área degradada e os resíduos adicionados na forma de massa verde após a semeadura do milho. Resíduos de leguminosas arbóreas influenciaram positivamente o crescimento de plantas de milho já aos 65 dias após a aplicação e favoreceram o acúmulo de nitrogênio, fósforo e potássio na parte aérea de plantas. Os adubos verdes utilizados melhoraram atributos químicos do solo logo aos 65 dias após a aplicação, com destaque para o N-nitrato (N-NO3 -), N-amônio (N-NH4 +), nitrogênio inorgânico total (N-NO3 - + N-NH4 +) e K, demonstrando que essas espécies são boas opções para recuperação de áreas degradadas no semiárido cearense.

Schistosoma mansoni tegument is a dynamic host-interactive layer that is an essential source of parasite antigens and a relevant field for schistosome vaccine research. Sm21.7 is a cytoskeleton antigen found in S. mansoni tegument that engenders protection in experimental challenge infection. Because of its crucial role in the parasite tegument and its promising protective capability, Sm21.7 is an exciting target for the development of therapeutic strategies. The present study describes Sm21.7 structural and biophysical features using circular dichroism spectroscopy and identifies linear B-cell epitopes of Sm21.7 using in-silico methods and immunoassay. The Sm21.7 gene was cloned into the pETDEST42 vector, and the recombinant protein was overexpressed in Escherichia coli DE3. The soluble protein was purified by affinity chromatography followed by ion-exchange chromatography. Purified recombinant Sm21.7 was analyzed by circular dichroism spectroscopy which demonstrated that the rSm21.7 structure was comprised of approximately 38% α-helices and its conformation remains stable at temperatures of up to 60 °C. Prediction of rSm21.7 B-cell epitopes was based on amino acid physicochemical properties. Sixteen peptides corresponding to predicted epitopes were synthesized and immunoreactivity assessed by spot peptide array using pooled rSm21.7-immunized mice sera or patients’ sera with different clinical forms of S. mansoni infection. Immunoassays revealed that sera from rSm21.7-immunized mice reacted predominantly with peptides located in the dynein-light chain domain (DLC) at the C-terminal region of rSm21.7. Comparative analysis of the antibody response of acute, intestinal and hepatosplenic patients’ sera to the Sm21.7 peptides showed that a differential recognition pattern of Sm21.7-derived peptides by intestinal patients’ sera might contribute to down-regulate the immune response in chronic intestinal patients. Together, the results may help the development of S. mansoni vaccine strategies based on the rSm21.7 antigen.

As startups, geralmente precisam de financiamento para crescer e estes novos empreendimentos frequentemente se voltam a investidores anjos como opção de capital. Investidores anjo são indivíduos com grande disponibilidade de recursos financeiros que decidem investir em negócios de risco trazendo frequentemente não somente capital, mas também conhecimento e experiência. O objetivo deste estudo é compreender melhor o processo decisório de investimento anjo em startups, identificando e analisando quais são os critérios de investimento priorizados pelos investidores anjo brasileiros em seus investimentos em startups. O ambiente principal de pesquisa deste trabalho foi a Gávea Angels, a mais antiga associação de anjos do Brasil, e de forma secundária, os investidores da rede de contatos dos pesquisadores. Em linha com grande parte dos estudos internacionais sobre o assunto, os resultados demonstraram que a despeito da importância dos critérios ligados ao mercado, produto e dados financeiros, os critérios priorizados foram aqueles ligados ao empreendedor e seu time de gestão, conforme segue: (1) confiabilidade/honestidade do empreendedor; (2) entusiasmo/comprometimento do empreendedor; (3) time de gestão; (4) expertise de domínio do empreendedor. Os critérios ?resiliência? e ?paixão do time? foram os atributos mais importantes na análise particular do ?time de gestão?

A zona costeira é uma área de grande importância econômica, principalmente devido ao turismo, pesca e às atividades portuárias. O desempenho de todas essas atividades, juntamente com a ocupação urbana desenfreada, faz com que seja necessária a aplicação de políticas ambientais mitigadoras e de zoneamento ambiental sustentável para que não haja a perda deste espaço. Nesse contexto, este estudo objetivou contextualizar as feições geomorfológicas do litoral de Santa Catarina por meio de observações de campo em seis municípios do Estado. Em todos os pontos visitados foi possível observar como a dinâmica costeira, incluindo os processos intempéricos atuantes, os agentes de transporte e a deposição do material sedimentar, influenciam na geomorfologia local. Foi observado diversos ambientes praiais em estágios erosivos, assim como algumas técnicas utilizadas na contenção destes processos. Assim, conclui-se que estudos observacionais são importantes para o conhecimento da evolução dos ambientes litorâneos, auxiliando no zoneamento e na gestão costeira.

Benzaldehyde and 2-hexanone are allomones produced by beagle dogs that reduce infestation by Rhipicephalus sanguineus sensu lato on these animals and on susceptible dogs which artificially release these repellents. These observations were obtained in previous laboratory tests or artificial infestations of susceptible dogs under controlled conditions. Here we evaluated the efficacy of collars delivering these repellents for suppressing the loads of R. sanguineus s.l. on naturally infested mixed-breed dogs under field conditions. Thirty dogs naturally infested with R. sanguineus s.l. were separated into two groups with 15 dogs each. The dogs from the treatment group received the collars with slow-release formulations of the allomones, and the dogs from the control group received collars without such compounds. Collar effectiveness tests were carried out over 30 days. All ticks found were removed from each dog, identified, and counted every collection day. The density of the different life stages of R. sanguineus s.l. varied greatly between groups and across evaluation times. Adult ticks were the stage most abundant and prevalent on infested dogs. The counts for larvae, nymphs and adults were similar between the control and the treatment groups within each tick collection day. Conversely, when we considered the total number of ticks over 30 days and gathered all life stages collected in the same animal, the dogs from the control group had significantly higher loads (35.3 ± 4.27) of ticks than the dogs from the treatment group (21.8 ± 2.96) (P0.01). In addition, the efficacy of treatment with repellent collars was high for adult ticks (30.8%) but was extremely low for reducing larval or nymphal infestation (0 or 2.6%, respectively). The present study demonstrates, for the first time, the efficacy of these volatile compounds on naturally infested dogs under field conditions. The development of a technologically enhanced device for slow release of the allomones tested here can be an advantageous alternative for reducing R. sanguineus s.l. infestation on dogs in residential environments.

O presente artigo aborda a desproporção existente quando comparado o número de unidades APACs femininas no Brasil com o número de unidades masculinas, analisando-se, ainda, a influência da desigualdade de gênero neste contexto de exclusão feminina. O desenvolvimento deste trabalho pauta-se em análise de dados provenientes do Sistema Nacional de Informações Penitenciárias (INFOPEN) e do site do Tribunal de Justiça de Minas Gerais (TJ/MG), onde há a descrição do número de APACs existentes no Brasil. A partir dos números obtidos, resta clara a exclusão da mulher, tanto no número de presídios femininos existentes, quanto no processo de implementação do Método Apaqueano. Busca-se, com o estudo, tratar da relação entre a desigualdade de gênero existente na sociedade brasileira e a invisibilização da mulher em situação de prisão, tratando das necessidades específicas do gênero feminino no âmbito prisional. Soma-se a isso, a inexistência de políticas públicas voltadas para um tratamento específico das reclusas, inviabilizando a garantia de reinserção social

The second enzyme of the naphthalene degradation pathway in Pseudomonas putida G7 is NahB, a dehydrogenase that converts cis-1,2-dihydroxy-1,2-dihydronaphthalene to 1,2-dihydroxynaphthalene. We report the cloning, optimization of expression, purification, kinetic studies and preliminary structural characterization of the recombinant NahB. The nahB gene was cloned into a T7 expression vector and the enzyme was overexpressed in Escherichia coli Rosetta (DE3) as an N-terminal hexa-histidine-tagged protein (6xHis-NahB). Using methods of enhancing protein stability in solution, we tested different expression, cell lysis, and purification protocols with and without ligand supplementation. The protein stability was evaluated by dynamic light scattering and circular dichroism spectroscopy assays. Best-derived protocols (expression at 18 °C, cell lysis with homogenizer, and three purification steps) were used to produce 20 mg of homogeneous 6xHis-NahB per liter of culture. The secondary and quaternary structures of 6xHis-NahB were assessed by circular dichroism and size-exclusion chromatography experiments, respectively. The enzyme was NAD+-dependent and active at pH 7.0 and 9.4 for the oxidation of the substrate. The Michaelis-Menten parameters determined at pH 7.0 and 25 °C for the substrate and cofactor, presented respective Km values of 6 and 350 μM, and a kcat value of 8.3 s−1. Furthermore, we identified conditions for the crystallization of 6xHis-NahB. X-ray diffraction data were collected from a single 6xHis-NahB crystal which diffracted to 2.21 A. The crystal belongs to space group I222, with unit-cell parameters a = 63.62, b = 69.50, and c = 117.47 A. The tertiary structure of 6xHis-NahB was determined using the molecular replacement method. Further structural refinement is currently underway.

Background Protein engineering has many applications for industry, such as the development of new drugs, vaccines, treatment therapies, food, and biofuel production. A common way to engineer a protein is to perform mutations in functionally essential residues to optimize their function. However, the discovery of beneficial mutations for proteins is a complex task, with a time-consuming and high cost for experimental validation. Hence, computational approaches have been used to propose new insights for experiments narrowing the search space and reducing the costs. Results In this study, we developed Proteus (an acronym for Protein Engineering Supporter), a new algorithm for proposing mutation pairs in a target 3D structure. These suggestions are based on contacts observed in other known structures from Protein Data Bank (PDB). Proteus’ basic assumption is that if a non-interacting pair of amino acid residues in the target structure is exchanged to an interacting pair, this could enhance protein stability. This trade is only allowed if the main-chain conformation of the residues involved in the contact is conserved. Furthermore, no steric impediment is expected between the proposed mutations and the surrounding protein atoms. To evaluate Proteus, we performed two case studies with proteins of industrial interests. In the first case study, we evaluated if the mutations suggested by Proteus for four protein structures enhance the number of inter-residue contacts. Our results suggest that most mutations proposed by Proteus increase the number of interactions into the protein. In the second case study, we used Proteus to suggest mutations for a lysozyme protein. Then, we compared Proteus’ outcomes to mutations with available experimental evidence reported in the ProTherm database. Four mutations, in which our results agree with the experimental data, were found. This could be initial evidence that changes in the side-chain of some residues do not cause disturbances that harm protein structure stability. Conclusion We believe that Proteus could be used combined with other methods to give new insights into the rational development of engineered proteins. Proteus user-friendly web-based tool is available at .

Pb27 antigen is an interesting alternative to immunological diagnosis of Paracoccidioidomycosis (PCM) and has demonstrated to be protective in experimental PCM. Its tertiary structure and possible function remained unknown till now. To study Pb27 at the atomic level, the recombinant protein was expressed in Escherichia coli BL21(DE3), purified, and its three-dimensional structure was solved by X-ray crystallography. Based on this structure, we performed a residue correlation analysis and in silico ligand search assays to address a possible biological function to Pb27. We identified Pb27 as a member of the extensive nucleotidyltransferase superfamily. The protein has an αβαβαβ topology with two domains (N- and C-terminal domains) and adopts a monomeric form as its biological unit in solution. Structural comparisons with similar members of the superfamily clearly indicate Pb27 C-terminal domain is singular and may play an important role in its biological function. Bioinformatics analysis suggested that Pb27 might bind to ATP and CTP. This suggestion is corroborated by the fact that a magnesium cation is coordinated by two aspartic acid residues present at the active site (between N- and C-terminal domains), as evidenced by X-ray diffraction data. Besides, NMR assays (1H-15N HSQC spectra) confirmed the binding of CTP to Pb27, demonstrating for the first time an interaction between a nucleotide and this protein. Moreover, we evaluated the reactivity of sera from patients with Paracoccidioides brasiliensis infection against the recombinant form of Pb27 and showed that it was recognized by sera from infected and treated patients. Predicted B and T cell epitopes were synthesized and further evaluated against sera of PCM patients, providing information of the most reactive peptides in Pb27 primary structure which interact with specific Pb27 antibodies.

A utilização de cultivares tolerantes à salinidade é uma das estratégias para a produção de alimentos em regiões afetadas pela salinização do solo. Objetivou-se com este estudo avaliar a tolerância de cultivares de pimentão ao estresse salino durante o processo de germinação das sementes e desenvolvimento das plântulas. O experimento foi conduzido em laboratório, adotando-se delineamento inteiramente aleatorizado, em esquema fatorial, com quatro repetições. O primeiro fator foi constituído por duas cultivares de pimentão (Casca Dura Ikeda e Amarelo SF 134); e o segundo, por cinco níveis de salinidade da água (0,0; 3,0; 6,0; 9,0 e 12,0 dS m-1). Avaliou-se a germinação, primeira contagem de germinação, índice de velocidade de germinação, tempo médio de germinação, comprimento de plântula, massa seca da raiz e parte aérea e índice de tolerância à salinidade. De forma geral, a salinidade da água promoveu efeitos deletérios na germinação e crescimento de plântulas das duas cultivares a partir da CE 6,0 dS m-1. A ‘Amarelo SF 134’ indica apresentar tolerância ao estresse salino no processo germinativo, enquanto a ‘Casca Dura Ikeda’, na fase de crescimento inicial de plântulas.

Study Design Clinical measurement study. Background Ankle sprain is a common condition. Therefore, translated, culturally adapted, and clinimetrically tested instruments for measuring the ankle are needed. Objectives To translate and cross-culturally adapt the Sports Athlete Foot and Ankle Score (SAFAS) and the global rating of change (GROC) into Brazilian Portuguese. This study also aimed to test the measurement properties of the SAFAS, the GROC, and the existing Brazilian-Portuguese versions of the numeric pain-rating scale (NPRS) and the ankle-hindfoot scale of the American Orthopaedic Foot and Ankle Society (AOFAS) in a group with ankle sprain. Methods The SAFAS and GROC instruments were cross-culturally adapted into Brazilian Portuguese and pretested in this population (n = 18). The measurement properties of the SAFAS, GROC, NPRS, and AOFAS (internal consistency, ceiling and floor effects, construct validity, reproducibility, and responsiveness) were tested in a test-retest design in patients with ankle sprain at baseline (n = 63) and at follow-up periods of 48 to 72 hours (n = 53) and 4 weeks (n = 43) after baseline. Results The SAFAS, GROC, NPRS, and AOFAS yielded considerable internal consistency (Cronbach alpha ranging from .76 to .97), moderate reliability (intraclass correlation coefficient model 2,1 ranging from 0.59 to 0.80), and dubious agreement (percentage of the standard error of measurement ranging from 13% to 22.43%). The highest correlations on construct validity were observed among the 4 SAFAS subscales (Pearson r = 0.75 to 0.86, P.01), but statistically significant values appeared in the intercorrelation of instruments. There were no observed floor or ceiling effects in any of the instruments. All of their scores of change showed moderate correlation with the GROC (r = -0.48 to 0.30, P.05) and could be classified as responsive. Conclusion The Brazilian-Portuguese versions of the SAFAS, GROC, NPRS, and AOFAS had acceptable measurement properties in high-performing athletes. J Orthop Sports Phys Ther 2016;46(12):1042-1050. Epub 8 Nov 2016. doi:10.2519/jospt.2016.6218.

Aims Control of diurnal Aedes aegypti with mycoinsecticides should consider the exposure of fungus-treated adults to sunlight, and especially to UV-B radiation that might affect activity of conidia applied on the mosquito′s surface. Methods and results Germination of M. anisopliae s.l. IP 46 conidia on SDAY medium was not affected at the lowest level of radiation with UV-B, 0.69 kJ m−2, but was retarded and reduced at higher 2.075 and 4.15 kJ m−2, and completely inhibited at ≥ 8.3 kJ m−2. In contrast, germination of conidia applied onto fiberglass nettings and exposed from 0–16.6 kJ m−2 did not differ significantly among among levels of irradiance exposure and the controls. There was also no significant impact of UV-B up to 16.6 kJ m−2 on adulticidal activity of IP and on the subsequent conidiogenesis on cadavers. The Quaite weighted UV-B irradiance in the laboratory (1152 mW m−2) was higher than the natural sunlight irradiance observed in the city of Goiânia in Central Brazil on midday (706 mW m−2 in August to 911 mW m−2 in October 2015). Conclusions UV-B does not impair the activity of IP 46 conidia applied previously to radiation on A. aegypti adults. Significance and Impact of Study Findings contribute to a better understanding of the effectiveness of M. anisopliae against day-active A. aegypti and its potential for biological mosquito control. This article is protected by copyright. All rights reserved.

In the present investigation we used a recombinant LiD1 toxin, named rLiD1his, from Loxosceles intermedia brown spider to elicit specific antibodies in mice carrying different Human Leukocyte Antigens class II (HLAII) {DRB1.0401 (DR4), DQB1.0601 (DQ6) and DQB1.0302 (DQ8)} as well as in BALB/C and C57BL/6 control mice. All mice strains produced high antibody titers against rLiD1his but DR4 mice antibodies (the lower responder mice) were not able to recognize L. intermedia crude venom. The anti-rLiD1his sera, except from DR4 mice, were able to neutralize dermonecrotic, hemorrhagic and edematogenic effects of rLiD1his in naïve rabbits. Overlapping peptides from the amino acid sequence of LiD1 toxin were prepared by SPOT method and differences in LiD1 epitope recognition were observed using different mice anti-rLiD1his sera. The region (160)DKVGHDFSGNDDISDVGK(177) was recognized by transgenic DQ8 and DQ6 mice sera. Other epitopes were recognized by at least two different animals' sera including (10)MGHMVNAIGQIDEFVNLG(27), (37)FDDNANPEYTYHGIP(51), (70)GLRSATTPGNSKYQEKLV(87) and (259)AAYKKKFRVATYDDN(273). Among these epitopes, the epitopes 37-51 and 160-177 have already been shown in previously studies as good candidates to be used alone or combined with other peptides to induce protective immune response against Loxosceles venoms. The results presented here highlight the importance of HLAII in antibody response and recognition of specific B-cell epitopes of rLiD1his spider toxin according to HLAII type and impact in the epitopic vaccine development against this spider.

Background Shoulder dislocation is a common injury among young athletes, especially those who play contact sports. Objective The aim of this systematic review is to investigate the return to preinjury sporting activities in patients who have undergone a conservative treatment with immobilisation after a primary anterior shoulder dislocation. Data sources We conducted electronic searches without restriction of language on the EMBASE and PUBMED databases, using the term ‘shoulder dislocation’ for each database. Study selection All prospective studies that have evaluated the role of immobilisation for first-time shoulder dislocation were included in this review. Synthesis methods and risk of bias Author9s name, year of publication, design of study, population, sample size, follow-up times, immobilisation method, and the rate of patients9 return to preinjury sporting activities were extracted from the selected articles. The instrument used for assessing the risk of bias of the included articles was the PEDro scale. Results A total of 7397 studies were found. From the four studies included in this review four of them used external rotation (ER) as treatment for three weeks. A total of 214 participants were recruited for ER and 169 for internal rotation (IR). On average, 58.9% (95% CI 51.1% to 64.7%) for immobilisation in ER and 42.6% (95% CI 34.5% to 49.8%) for immobilisation in IR returned to preinjury sports activities. PEDro scores for the included studies ranged from 3 to 6. Conclusions On the basis of the best available evidence that the studies reported, on average

Human milk oligosaccharide (HMO) composition varies throughout lactation and can be influenced by maternal characteristics. This study describes HMO variation up to three months postpartum and explores the influences of maternal sociodemographic and anthropometric characteristics in a Brazilian prospective cohort. We followed 101 subjects from 28&ndash, 35 gestational weeks (baseline) and throughout lactation at 2&ndash, 8 (visit 1), 28&ndash, 50 (visit 2) and 88&ndash, 119 days postpartum (visit 3). Milk samples were collected at visits 1, 2 and 3, and 19 HMOs were quantified usinghigh-performance liquid chromatography with fluorescence detection (HPLC-FL). Friedman post-hoc test, Spearman rank correlation for maternal characteristics and HMOs and non-negative matrix factorization (NMF) were used to define the HMO profile. Most women were secretors (89.1%) and presented high proportion of 2&rsquo, fucosyllactose (2ꞌFL) at all three sample times, while lacto-N-tetraose (LNT, 2&ndash, 8 days) and lacto-N-fucopentaose II (LNFPII, 28-50 and 88-119 days) were the most abundant HMOs in non-secretor women. Over the course of lactation, total HMO weight concentrations (g/L) decreased, but total HMO molar concentrations (mmol/L) increased, highlighting differential changes in HMO composition over time. In addition, maternal pre-pregnancy body mass index (BMI) and parity influence the HMO composition in healthy women in this Brazilian cohort.

Capsaicin, an agonist of TRPV1, evokes intracellular [Ca2+] transients and glutamate release from perfused trigeminal ganglion. The spider toxin PnTx3-5, native or recombinant is more potent than the selective TRPV1 blocker SB-366791 with IC50 of 47 ± 0.18 nM, 45 ± 1.18 nM and 390 ± 5.1 nM in the same experimental conditions. PnTx3-5 is thus more potent than the selective TRPV1 blocker SB-366791. PnTx3-5 (40 nM) and SB-366791 (3 μM) also inhibited the capsaicin-induced increase in intracellular Ca2+ in HEK293 cells transfected with TRPV1 by 75 ± 16% and 84 ± 3.2%, respectively. In HEK293 cells transfected with TRPA1, cinnamaldehyde (30 μM) generated an increase in intracellular Ca2+ that was blocked by the TRPA1 antagonist HC-030031 (10 μM, 89% inhibition), but not by PnTx3-5 (40 nM), indicating selectivity of the toxin for TRPV1. In whole-cell patch-clamp experiments on HEK293 cells transfected with TRPV1, capsaicin (10 μM) generated inward currents that were blocked by SB-366791 and by both native and recombinant PnTx3-5 by 47 ± 1.4%; 54 ± 7.8% and 56 ± 9.0%, respectively. Intradermal injection of capsaicin into the rat left vibrissa induced nociceptive behavior that was blocked by pre-injection with either SB-366791 (3 nmol/site i.d., 83.3 ± 7.2% inhibition) or PnTx3-5 (100 fmol/site, 89 ± 8.4% inhibition). We conclude that both native and recombinant PnTx3-5 are potent TRPV1 receptor antagonists with antinociceptive action on pain behavior evoked by capsaicin.

Salicylate hydroxylase (NahG) is a flavin-dependent monooxygenase that catalyzes the decarboxylative hydroxylation of salicylate into catechol in the naphthalene degradation pathway in Pseudomonas putida G7. We explored the mechanism of action of this enzyme in detail using a combination of structural and biophysical methods. NahG shares many structural and mechanistic features with other versatile flavin-dependent monooxygenases, with potential biocatalytic applications. The crystal structure at 2.0 A resolution for the apo form of NahG adds a new snapshot preceding the FAD binding in flavin-dependent monooxygenases. The kcat/Km for the salicylate reaction catalyzed by the holo form is >105 M−1 s−1 at pH 8.5 and 25 °C. Hammett plots for Km and kcat using substituted salicylates indicate change in rate-limiting step. Electron-donating groups favor the hydroxylation of salicylate by a peroxyflavin to yield a Wheland-like intermediate, whereas the decarboxylation of this intermediate is faster for electron-withdrawing groups. The mechanism is supported by structural data and kinetic studies at different pHs. The salicylate carboxyl group lies near a hydrophobic region that aids decarboxylation. A conserved histidine residue is proposed to assist the reaction by general base/general acid catalysis.

The purpose of this study was to investigate the effectiveness of adding photobiomodulation therapy and neuromuscular electrical stimulation (NMES) to volleyball athletes’ training, focusing on muscle strength and jumping skills. Thirty-six athletes were randomly placed into three groups: control, photobiomodulation therapy, and NMES. The athletes trained to improve their muscle strength and jumping skills. The athletes in the photobiomodulation therapy group were submitted to photobiomodulation therapy (850 nm, continuous, energy density 0.8 J/cm2, radiant energy per point 6 J, total radiant energy 36 J) before undergoing strength and plyometric training. The NMES group additionally underwent NMES-based quadriceps femoris muscle strength training (base frequency 1 kHz, frequency modulation 70 Hz, intensity maximum tolerable). The variables analyzed were muscle strength, jumping ability, global impression, and jump frequency; they were measured at baseline and during follow-ups at 6 and 8 weeks. The statistical analysis was conducted on an intention-to-treat basis. The between-group differences and their respective 95% CIs were calculated using linear mixed models by using group, time, and group-versus-time interaction terms. Dominant lower limb strength improved the most in the NMES group compared to the control group (mean difference = 1.4, 95% CI = .5 to 2.4). Non-dominant lower limb strength increased in both the photobiomodulation therapy group (mean difference = 1.1, 95% CI = .3 to 2) and the NMES group (mean difference = 1.9, 95% CI = 1.1 to 2.8) compared to the control group, but the NMES group improved more than the photobiomodulation therapy group (mean difference = 0.8, 95% CI = 0.1 to 1.7). The NMES group had the greatest improvement in global perceived effect scale compared to the control group (mean difference = 1.1, 95% CI = 1 to 2.2). Dominant lower limb strength improved in the NMES group compared to the control group. Non-dominant lower limb strength increased in both the photobiomodulation therapy group and the NMES group compared to the control group, but the NMES group improved significantly more than the photobiomodulation therapy group; the NMES group also improved in the global perceived effect scale compared to the control group. This study found that, for volleyball athletes, photobiomodulation therapy and NMES both promoted benefits in terms of muscle-strength gain. In addition, these benefits were maintained for 2 weeks even after training was interrupted. Dominant lower limb strength improved in the NMES group compared to the control group. Non-dominant lower limb strength increased in both the photobiomodulation therapy group and the NMES group compared to the control group, but the NMES group improved significantly more than the photobiomodulation therapy group; the NMES group also improved in global impression of jumps compared to the control group.

In Trypanosoma cruzi, the etiologic agent of Chagas disease, Rad51 (TcRad51) is a central enzyme for homologous recombination. Here we describe the different roles of TcRad51 in DNA repair. Epimastigotes of T. cruzi overexpressing TcRAD51 presented abundant TcRad51-labeled foci before gamma irradiation treatment, and a faster growth recovery when compared to single-knockout epimastigotes for RAD51. Overexpression of RAD51 also promoted increased resistance against hydrogen peroxide treatment, while the single-knockout epimastigotes for RAD51 exhibited increased sensitivity to this oxidant agent, which indicates a role for this gene in the repair of DNA oxidative lesions. In contrast, TcRad51 was not involved in the repair of crosslink lesions promoted by UV light and cisplatin treatment. Also, RAD51 single-knockout epimastigotes showed a similar growth rate to that exhibited by wild-type ones after treatment with hydroxyurea, but an increased sensitivity to methyl methane sulfonate. Besides its role in epimastigotes, TcRad51 is also important during mammalian infection, as shown by increased detection of T. cruzi cells overexpressing RAD51, and decreased detection of single-knockout cells for RAD51, in both fibroblasts and macrophages infected with amastigotes. Besides that, RAD51-overexpressing parasites infecting mice also presented increased infectivity and higher resistance against benznidazole. We thus show that TcRad51 is involved in the repair of DNA double strands breaks and oxidative lesions in two different T. cruzi developmental stages, possibly playing an important role in the infectivity of this parasite., Author summary Trypanosoma cruzi is the causative agent of Chagas disease, a tropical neglected illness that affects 6 million people worldwide, mostly in Latin America. Our research group focuses on the elucidation of DNA repair and metabolism of T. cruzi, and on the possible implications of those mechanisms in both parasite genetic diversity and Chagas disease development. In this work, we investigated the involvement of homologous recombination in the oxidation-induced damage response, and in DNA repair, in T. cruzi cells after gamma radiation exposure. We also examined whether TcRad51 –a key protein for homologous recombination–could be an important factor for T. cruzi’s infectivity. We generated genetically-modified T. cruzi cells for RAD51 and studied the in vitro and in vivo infectivity of these mutant cells. Our results indicate that TcRad51 is a key protein involved in the repair of oxidative and DNA double-strand breaks lesions in two crucial developmental forms of T. cruzi.

There is a natural duality between line congruences in $\mathbb{R}^3$ and surfaces in $\mathbb{R}^4$ that sends principal lines into asymptotic lines. The same correspondence takes the discriminant curve of a line congruence into the parabolic curve of the dual surface. Moreover, it takes the ridge curves to the flat ridge curves, while the subparabolic curves of a line congruence are taken to certain curves on the surface that we call flat subparabolic curves. In this paper, we discuss these relations and describe the generic behavior of the subparabolic curves at the discriminant curve of the line congruence, or equivalently, the parabolic curve of the dual surface. We also discuss Loewner's conjectures under the duality viewpoint., Comment: 16 pages, 2 figures

Teses defendidas no 1º/2015.

Objective: To compare the performance and effectiveness of a hydrocolloid dressing (HD) and a transparent polyurethane film (PF) in preventing pressure ulcer (PU) development. Method: The study was conducted in the intensive care unit, coronary care unit and medical clinic of the Holy House of Mercy of Passos, Brazil. Data were collected 48 hours after admission and during hospitalisation. The Braden scale was used for PU risk assessment. Consecutive eligible patients without PUs were randomly assigned by lottery to the two groups, either the HD or PF group. Results: Of the 160 eligible patients, significant between-group differences were found in the mean total number of dressing changes (HD, 6.09 ± 1.655 changes; PF, 5.59 ± 2.036 changes; p=0.010), and mean number of dressing changes in the sacral region (HD, 2.50 ± 0.871; PF, 2.05 ± 0.825; p=0.001), with the PF group requiring significantly fewer changes than the HD group. The most common reasons for changing dressings in both groups were moisture (PF 51.1%; HD 47.9%) and shear (HD 43%; PF 38.9%), with a significant difference in shear between groups. The incidence of PUs was significantly lower (p=0.038) in the PF group (8.7%) compared with that in the HD group (15%). Conclusion: The results suggest that the transparent polyurethane film had a better performance and was more effective than the hydrocolloid dressing in preventing PU development.

There are several methods used in sports science to identify asymmetries in athletes, given their purported relevance to injury prevention and performance optimization. We aimed to verify whether asymmetries provided by isokinetic assessments, jump tests, and tensiomyography (TMG) are associated with each other, and whether their respective functional indices are related to jumping ability. TMG parameters, unilateral and bilateral squat-jump (SJ) and countermovement-jump (CMJ) performances, and peak torque in knee-extension and flexion with angular velocities of 60o/s and 300o/s for twenty-four soccer players were retained for analyses. Asymmetry was detected by examining the percentage difference between dominant and non-dominant legs. The median-split technique was used to identify the best and worst performers in SJ and CMJ tests. Results revealed that the asymmetries detected in the three different methods were not interrelated. Curiously, better performances in SJ and CMJ tests were associated with higher asymmetry levels. Furthermore, only the knee-extension peak torque at both angular velocities was correlated moderately to largely (r=0.48-0.66) with jump performance. Despite their recognized ability to predict the risk of injury, the absence of interrelationships between TMG, isokinetic tests, and unilateral jumps precludes their single use as a unique functional screening diagnostic. Finally, and very importantly, lower-limb asymmetry is not necessarily related to impaired vertical jump performance in soccer players.

The need to insert the durability factor on dimensioning concrete structures has been stimulating the development of new models to predict the service life of those structures. Among those new models, the use of data from Non-Destructive Testing (NDT) is recommended. It is known that electrical resistivity is a parameter related with permeability and, in consequence, with the resistance of the concrete against the intrusion of aggressive agents. Electrical resistivity may be monitored with time by a NDT method, by a low cost and easy execution technique. However, some parameters of this essay need to be further discussed in the literature aiming to standardize this technique. Thus, this study had the goal to verify the influence of test specimen’s shape in measurements of electrical resistivity. In order to that, cylindrical and prismatic test specimens have been molded from a same type of concrete and electrical resistivity was monitored during a period of 120 days. Results show significant differences between the values obtained. Test specimens in cylindrical shape had the higher results. A good correlation between factors was found with R2 higher than 0.94.

Estudo transversal longitudinal com o objetivo de verificar causas de não doação de órgãos e tecidos entre doadores potenciais abordados por Comissão de Transplantes. Foram analisados 573 registros de abordagem a doadores potenciais, no perí­odo de janeiro de 2007 aoutubro de 2008, com média global mensal de 26,10 casos/mês. A faixa etária com maior incidência de negativa de doação foi acima de 71 anos (22,34%). A doação de tecidos apresentou 81,33% de anuências com aumento crescente do total geral de 2007 para 2008. Anão doação ocorreu por recusa de familiares (6,80%), problemas logí­sticos e estruturais (0,17%), contraindicação médica (89,86%). Os doadores potenciais localizavam-se no centro cirúrgico (0,70%); home care (0,17%); pronto atendimento (4,19%); residência (0,17%); setor de internação (28,62%); Unidade de Terapia Intensiva (65,10%). Concluiu-se que as causas de não doação estão relacionadas com a idade dos potenciais doadores e com as patologias associadas a estes indiví­duos. Enfatiza-se a Unidade de Terapia Intensiva como setor com maior potencial de doação, exigindo preparo dos profissionais do setor, com ênfase no médico, por ser o profissional com maior ascendência sobre a famí­lia do doador.Palavras-chave: conhecimentos, atitudes e prática em saúde, transplante de órgãos, doação dirigida de tecido.

In human and murine models strongyloidiasis induce a Th2 type response. In the current study we investigated the role of different loads of Strongyloides venezuelensis in the immune response raised against the parasite and the participation of the major histocompatibility complex (MHC) class II molecule in the disease outcome in face of the different parasite burden. The C57BL/6 wild type (WT) and MHC II−/− mice were individually inoculated by subcutaneous injection with 500 or 3000 S. venezuelensis L3. The MHC II−/− mice infected with 3000L3 were more susceptible to S. venezuelensis infection when compared with WT groups, in which the parasite was completely eliminated. The production of Th2 cytokines and specific IgG1 or IgE antibodies against parasite were significantly lowered in MHC II−/− infected mice with different larvae inoculums. The infection of MHC II−/− mice with S. venezuelensis induced slight inflammatory alterations in the small intestine, and these lesions were lower when compared with WT mice, irrespective of the parasite load utilized to infect animals. Finally, we concluded that MHC class II molecules are essential in the immune response against S. venezuelensis mainly when infection occurs with high parasite inoculum.

The salicylaldehyde dehydrogenase (NahF) catalyzes the oxidation of salicylaldehyde to salicylate using NAD(+) as a cofactor, the last reaction of the upper degradation pathway of naphthalene in Pseudomonas putida G7. The naphthalene is an abundant and toxic compound in oil and has been used as a model for bioremediation studies. The steady-state kinetic parameters for oxidation of aliphatic or aromatic aldehydes catalyzed by 6xHis-NahF are presented. The 6xHis-NahF catalyzes the oxidation of aromatic aldehydes with large kcat/Km values close to 10(6) M(-1) s(-1). The active site of NahF is highly hydrophobic, and the enzyme shows higher specificity for less polar substrates than for polar substrates, e.g., acetaldehyde. The enzyme shows α/β folding with three well-defined domains: the oligomerization domain, which is responsible for the interlacement between the two monomers; the Rossmann-like fold domain, essential for nucleotide binding; and the catalytic domain. A salicylaldehyde molecule was observed in a deep pocket in the crystal structure of NahF where the catalytic C284 and E250 are present. Moreover, the residues G150, R157, W96, F99, F274, F279, and Y446 were thought to be important for catalysis and specificity for aromatic aldehydes. Understanding the molecular features responsible for NahF activity allows for comparisons with other aldehyde dehydrogenases and, together with structural information, provides the information needed for future mutational studies aimed to enhance its stability and specificity and further its use in biotechnological processes.

Two xylanases from Aspergillus nidulans, XlnB and XlnC, were expressed in Pichia pastoris, purified and characterized. XlnB and XlnC achieved maximal activities at 60 °C and pH 7.5 and at 50 °C and pH 6.0, respectively. XlnB showed to be very thermostable by maintaining 50% of its original activity after 49 h incubated at 50 °C. XlnB had its highest activity against wheat arabinoxylan while XlnC had the best activity against beechwood xylan. Both enzymes were completely inhibited by SDS and HgCl2. Xylotriose at 1 mg/ml also totally inibited XlnB activity. TLC analysis showed that the main product of beechwood xylan hydrolysis by XlnB and XlnC was xylotetraose. An additive effect was shown between XlnB and XlnC and the xylanases of two tested commercial cocktails. Sugarcane bagasse saccharification results showed that these two commercial enzymatic cocktails were able to release more glucose and xylose after supplementation with XlnB and XlnC.

O presente estudo foi realizado com o objetivo de investigar estratégias de uso de água salobra sobre o acúmulo de matéria seca e nutrientes, duas cultivares de melão (Sancho- C1 e Medellín- C2) irrigadas com água de baixa (S1 = 0,61 dS m-1) e alta (S2 = 4,78 dS m-1) salinidade por fase da cultura: S1S1S2S2- T1, S2S1S2S2- T2, S2S2S1S2- T3 (os 1, 2, 3 e 4º termos correspondem, respectivamente, às fases de crescimento, floração, crescimento dos frutos e maturação e colheita) e, ainda a irrigação alternada, durante todo o ciclo, com água S1 por 2 dias consecutivos seguido com água S2 por um dia (S12dias + S21dia- T4) e a irrigação com água S2 durante todo o ciclo- T5. Além disto foi utilizada, como testemunha, a irrigação com a mistura de 37% da água S1 e 63% da água S2- T0 (manejo adotado atualmente na fazenda em que o experimento foi conduzido). O delineamento experimental utilizado foi o de blocos inteiramente casualizados em um esquema de parcelas subdivididas 6 x 2 (manejo da água salina x cultivar) com quatro repetições. O acúmulo de fitomassa seca total na parte aérea da cultivar Sancho foi superior ao de Medellín, em todas as estratégias de manejo utilizado; os nutrientes mais absorvidos pelas plantas das duas cultivares foram o cálcio, o potássio e o nitrogênio.

Este trabalho foi realizado com o objetivo de avaliar o efeito de diferentes níveis de salinidade da água de irrigação e tratamento de sementes com regulador vegetal no desenvolvimento do algodoeiro. O delineamento experimental adotado foi inteiramente ao acaso, arranjados em esquema fatorial 5 x 2 com quatro repetições. Os tratamentos resultaram da combinação de cinco níveis de salinidade da água de irrigação (S1-0,5; S2-2,0; S3-3,5; S4-5,0 e 6,5 dS m-1) em sementes tratadas e não tratadas com regulador vegetal. As características avaliadas foram: número de folhas, área foliar, altura, massa seca do caule, das folhas e da parte vegetativa, área foliar específica e razão de área foliar. Houve interação entre salinidade e tratamentos de sementes com cloreto de mepiquat apenas para área foliar e razão de área foliar. Todas as demais características foram reduzidas com o aumento da salinidade da água de irrigação, com maiores reduções para área foliar (média de 65,8%) e massa seca da parte vegetativa (64%). O tratamento das sementes com cloreto de mepiquat afetou o desenvolvimento das plantas, independente da salinidade.

CMS1MS2 (CC-Ib) from Carica candamarcensis (Vasconcellea cundinamarcensis) is a cysteine proteinase found as a single polypeptide containing 213 residues of 22,991 Da. The enzyme was purified by three chromatographic steps, two of them involving cationic exchange. Crystals of CMS1MS2 complexed with E-64 were obtained by the hanging drop vapor-diffusion method at 291 K using ammonium sulfate and polyethylene glycol 4000/8000 as precipitant. The complex CMS1MS2-E-64 crystallized in the tetragonal space group P4(1)2(1)2 with unit-cell parameters; a = b = 73.64, c = 118.79 Å. The structure was determined by Molecular Replacement and refined at 1.87 Å resolution to a final R factor of 16.2 % (R (free) = 19.3 %). Based on the model, the structure of CMS1MS2 (PDB 3IOQ) ranks as one of the least basic cysteine isoforms from C. candamarcensis, is structurally closer to papain, caricain, chymopapain and mexicain than to the other cysteine proteinases, while its activity is twice the activity of papain towards BAPNA substrate. Two differences, one in the S2 subsite and another in the S3 subsite of CMS1MS2 may contribute to the enhanced activity relative to papain. In addition, the model provides a structural basis for the sensitivity of CMS1MS2 to inhibition by cystatin, not shown by other enzymes of the group, e.g., glycyl endopeptidase and CMS2MS2.

Background Several forms of conservative treatment have been the focus of many recent studies in knee osteoarthritis (OA). Among these techniques, the application of pulsed shortwave (PSW) treatment has been widely used, but the optimal dose and application time have not been well established. Objective The purposes of this study were: (1) evaluate the effect of PSW treatment in different doses and (2) to compare low-dose and high-dose PSW groups with control and placebo groups. Design This was a randomized clinical trial. Setting The study was conducted in the physical therapy department of 2 large urban hospitals. Patients One hundred twenty-one women (mean age=60 years, SD=9) with a diagnosis of knee OA participated in the study. Intervention and Measurements Participants were distributed randomly into 4 groups: 35 participants did not receive any treatment (control group), 23 received a placebo treatment, 32 received low-dose PSW treatment (power of 14.5 W, treatment duration of 19 minutes, and total energy of 17 kJ), and 31 received high-dose PSW treatment (power of 14.5 W, treatment duration of 38 minutes, and total energy of 33 kJ). An 11-point numerical pain rating scale and the Knee Osteoarthritis Outcome Score were used to assess pain and function in 3 stages: at initial evaluation (pretreatment), immediately after treatment, and at 12-month follow-up. Results The 4 groups were homogeneous prior to treatment with respect to demographics, pain, and functional scale data. The results demonstrated the short-term effectiveness of the PSW at low and high doses in patients with knee OA. Both treatment groups showed a significant reduction in pain and improvement in function compared with the control and placebo groups (effect size: range=20.0–23.4 for the low-dose PSW group and range=15.7–16.5 for the high-dose PSW group). There were no differences in results between PSW doses, although a low dose of PSW appeared to be more effective in the long term. Limitations These results were achieved without physical exercise, which could have positively influenced the results. Conclusions Pulsed shortwave treatment is an effective method for pain relief and improvement of function and quality of life in the short term in women with knee OA. On the basis of the results, application of PSW treatment is recommended in the female population with knee OA. However, conclusions regarding the 12-month follow-up should be analyzed carefully due to the high dropout rate.

Resumen pt: O estresse hidrico e salino sao os principais estresses abioticos que limitam a produtividade das culturas em todo o mundo. A reducao na disponibilidade ...

Resumen pt: O trabalho foi desenvolvido com o objetivo de avaliar o efeito da salinidade e de diferentes fontes de nitrogenio no desenvolvimento inicial do girassol....

O uso de água salina, na irrigação, tem sido um desafio para os pesquisadores e produtores rurais, sendo constantemente desenvolvidos estudos que possibilitem o uso de água, de qualidade inferior, sem afetar o rendimento e qualidade dos produtos agrícolas. Este trabalho foi realizado com o objetivo de avaliar o efeito de diferentes níveis de salinidade da água de irrigação e doses de fósforo na cultura do rabanete. O delineamento experimental adotado foi inteiramente ao acaso, arranjados em esquema fatorial 4 x 4 com três repetições. Os tratamentos resultaram da combinação de quatro níveis de salinidade da água de irrigação (S1-0,5; S2-2,0; S3-3,5 e S4-5,0 dS m-1) com quatro doses de fósforo (0; 100; 200 e 300 mg dm-3). As variáveis avaliadas foram: número de folhas, área foliar, massa fresca e seca das raízes. A cultura do rabanete respondeu significativamente a interação entre salinidade e fósforo para área foliar, massa fresca e seca das raízes; e de forma isolada para número de folhas. Doses crescentes de fósforo reduziram à sensibilidade da cultura do rabanete à salinidade de até 3,5 dS m-1. O manejo da adubação fosfatada é uma alternativa para amenizar o efeito da salinidade no desenvolvimento da cultura do rabanete.