Population genetic structuring in pacu (Piaractus mesopotamicus) across the Paraná-Paraguay basin: evidence from microsatellites

The Paraná-Paraguay basin encompasses central western Brazil, northeastern Paraguay, eastern Bolivia and northern Argentina. The Pantanal is a flooded plain with marked dry and rainy seasons that, due to its soil characteristics and low declivity, has a great water holding capacity supporting abundant fish fauna. Piaractus mesopotamicus, or pacu, endemic of the Paraná-Paraguay basin, is a migratory species economically important in fisheries and ecologically as a potential seed disperser. In this paper we employ eight microsatellite loci to assess the population structure of 120 pacu sampled inside and outside the Pantanal of Mato Grosso. Our main objective was to test the null hypothesis of panmixia and to verify if there was a different structuring pattern between the Pantanal were there were no physical barriers to fish movement and the heavily impounded Paraná and Paranapanema rivers. All loci had moderate to high levels of polymorphism, the number of alleles varied from three to 18. The average observed heterozygosity varied from 0.068 to 0.911. After the Bonferroni correction three loci remained significant for deviations from Hardy-Weinberg, and for those the frequency of null alleles was estimated. F ST and R ST pairwise comparisons detected low divergence among sampling sites, and differentiation was significant only between Paranapanema and Cuiabá and Paranapanema and Taquari. No correlation between genetic distance and the natural logarithm of the geographic distance was detected. Results indicate that for conservation purposes and for restoration programs small genetic differences detected in the Cuiabá and Paranapanema rivers should be taken in consideration.

Determinação eletroquímica da capacidade antioxidante de sucos de frutas industrializados usando o CRAC assay

Fruits juices are natural sources of several compounds that present antioxidant action. Together with the fruits, they contribute with almost 40% of the antioxidant capacity in a healthy diet avoiding and preventing diseases deriving from oxidative stress. The present study determined the antioxidant capacity of seven samples of industrialized fruits juices applying CRAC (Ceric Reducing/Antioxidant Capacity) assay, a new electrochemistry assay that evaluates, by means of chronoamperometric measurements, the ability of a sample in reducing species Ce4+ in acid media. At the end of the assay was obtained the following classification: cashew > guava > grape > mango > apple > orange > passion fruit.

Vitaminas e minerais com propriedades antioxidantes e risco cardiometabólico: controvérsias e perspectivas

No processo celular de obtenção de energia, são gerados compostos chamados espécies reativas de oxigênio (ERO) que, em excesso, podem causar danos celulares. Estresse oxidativo resulta do desequilíbrio no estado de óxido-redução a favor da oxidação. Dos mecanismos de defesa antioxidante, participam enzimas endógenas e algumas vitaminas e minerais. A vitamina E encontra-se no plasma e na partícula de LDL, protegendo lipídeos da oxidação. Estudos observacionais relataram associação inversa entre ingestão de vitamina E e risco cardiometabólico (RCM). Entretanto, ensaios clínicos não comprovaram a eficácia de sua suplementação nos desfechos cardiometabólicos. A vitamina C participa do sistema de regeneração da vitamina E, mantendo o potencial antioxidante plasmático. Dados sobre os benefícios de sua suplementação na redução do risco cardiometabólico são inconclusivos. A atividade antioxidante dos carotenoides é responsável, em parte, por seu papel protetor contra doenças cardiovasculares e cânceres. A suplementação desse nutriente também não trouxe resultados consistentes no que se refere à redução do RCM. A participação do zinco e do selênio na defesa antioxidante vem sendo estudada mais recentemente, mas a sua suplementação em indivíduos com níveis séricos normais e ingestão adequada na dieta desses minerais não parece ser necessária. De um modo geral, há muita controvérsia sobre o papel desses micronutrientes no RCM. Estudos epidemiológicos sugerem que o consumo de substâncias antioxidantes provenientes da dieta ou dietas ricas em frutas e hortaliças diminui o RCM. Mais estudos são necessários antes de se recomendar o uso de antioxidantes isolados na forma de suplementos para tal finalidade.

Aspectos atuais sobre aminoácidos de cadeia ramificada e exercício físico

Em humanos saudáveis, nove aminoácidos são considerados essenciais, uma vez que não podem ser sintetizados endogenamente e, portanto, devem ser ingeridos por meio da dieta. Dentre os aminoácidos essenciais, se incluem os três aminoácidos de cadeia ramificada, ou seja, leucina, valina e isoleucina. Esses aminoácidos participam da regulação do balanço protéico corporal além de serem fonte de nitrogênio para a síntese de alanina e glutamina. No tocante à regulação da síntese protéica muscular, verifica-se que a leucina age estimulando a fase de iniciação da tradução do RNA-mensageiro em proteína, por mecanismos tanto dependentes quanto independentes de insulina. No que concerne ao exercício físico, supõe-se que esses aminoácidos estejam envolvidos na fadiga central, no balanço protéico muscular, na secreção de insulina, na modulação da imunocompetência, no aumento da performance de indivíduos que se exercitam em ambientes quentes e na diminuição do grau de lesão muscular. Nesse contexto, essa revisão aborda os aspectos atuais do metabolismo e da suplementação de aminoácidos de cadeia ramificada no exercício físico.

Entre a frieza, o cálculo e a "vida loka": violência e sofrimento no trajeto de um adolescente em cumprimento de medida socioeducativa

O conceito de sofrimento social caracteriza-se pela compreensão das situações de aflição e dor como experiências sociais e não como problemas individuais. Este trabalho analisa a natureza social e política do sofrimento de um adolescente em cumprimento de medida socioeducativa. Inspirado na abordagem de Veena Das, o artigo se apóia em "carne" e discurso para problematizar a relação entre cidadania e segmentos juvenis discriminados, que se manifesta nas ambiguidades das práticas institucionais presentes no fluxo de execução de medidas socioeducativas. O artigo analisa as contradições entre o objetivo institucional de evitar a reincidência de atos infracionais, auxiliando o adolescente a tornar-se um cidadão autônomo, e as narrativas e expressões corporais dos adolescentes durante o cumprimento das medidas. A trajetória aqui descrita leva ao reconhecimento de que o trânsito da medida de internação para as medidas em meio aberto se dá sob a tensão entre o discurso institucional de reorganizar a vida escolar, familiar e comunitária e a experiência cotidiana dos adolescentes, que segue marcada pela constante ameaça policial e pela privação de acessos a bens públicos. O cumprimento de medidas socioeducativas acaba por reforçar entre os adolescentes a aflição de serem socialmente tidos como suspeitos e fugitivos e, consequentemente, a incorporação de um lugar social particular, o de membro do "mundo do crime". O desempenho na vida cotidiana de um "estilo bandido" revela formas de resposta ao discurso dominante no sistema socioeducativo, contexto que indica o paradoxo do Estado brasileiro, que garante uma democracia formal enquanto viola direitos civis.

Produção in vitro de embriões de ovinos: uma visão crítica do método e de seu resultado a campo

A metodologia de produção in vitro de embriões de ovinos implica no desenvolvimento de meios de maturação, fertilização e cultivo que permitam aumentar a taxa de clivagem e desenvolvimento, tanto para o investimento biotecnológico em programas comerciais, quanto para sua utilização em clonagem e transgenia dessa espécie animal. Do ponto de vista da pesquisa, os ovócitos podem ser obtidos pelas técnicas de punção e slicing a partir de ovários oriundos de matadouros, ou através de aspiração folicular por laparoscopia. Como vantagem, este método permite o uso de uma mesma doadora estimulada hormonialmente em intervalos periódicos, mantida sob rigoroso controle sanitário, o que é de vital importância para a produção de biofármacos em programas que utilisem os ovinos como modelo biológico. Por outro lado, em nosso país a demanda pela multiplicação de animais de alto valor genético, seja pela produtividade ou pelo elevado valor comercial dos mesmos, impõe o desenvolvimento, adaptação e otimização das diferentes metodologias desenvolvidas ao longo dos ultimos anos em laboratórios de referência mundiais. Nesse contexto, cresce de importância o perfeito conhecimento da fisiologia dessa espécie e das raças criadas em nosso país, e da problemática da produção in vitro de seus embriões. Respeitando essas premissas, gerar o desenvolvimento de protocolos que permitam não apenas aumentar a população de ovócitos passíveis de maturação in vitro, mas de sua competência ao desenvolvimento ao estágio de blastocisto, ou, alternativamente, sua transferência a receptoras em estágios precoces do desenvolvimento, evitando assim as conhecidas perdas durante o desenvolvimento in vitro, e o baixo percentual de gestações que chegam a termo, com cordeiro saudáveis. Trata-se de um desafio, que já apresenta os primeiros resultados em nosso país, tanto na produção comercial de embriões produzidos in vitro, quanto em programas de clonagem e transgenia.

Genetic diversity of turmeric germplasm (Curcuma longa; Zingiberaceae) identified by microsatellite markers

Turmeric (Curcuma longa) is a triploid, vegetatively propagated crop introduced early during the colonization of Brazil. Turmeric rhizomes are ground into a powder used as a natural dye in the food industry, although recent research suggests a greater potential for the development of drugs and cosmetics. In Brazil, little is known about the genetic variability available for crop improvement. We examined the genetic diversity among turmeric accessions from a Brazilian germplasm collection comprising 39 accessions collected from the States of Goias, Mato Grosso do Sul, Minas Gerais, Sao Paulo, and Para. For comparison, 18 additional genotypes were analyzed, including samples from India and Puerto Rico. Total DNA was extracted from lyophilized leaf tissue and genetic analysis was performed using 17 microsatellite markers (single-sequence repeats). Shannon-Weiner indexes ranged from 0.017 (Minas Gerais) to 0.316 (Sao Paulo). Analyses of molecular variance (AMOVA) demonstrated major differences between countries (63.4%) and that most of the genetic diversity in Brazil is found within states (75.3%). Genotypes from Sao Paulo State were the most divergent and potentially useful for crop improvement. Structure analysis indicated two main groups of accessions. These results can help target future collecting efforts for introduction of new materials needed to develop more productive and better adapted cultivars.

Variations on a theme: diversification of cuticular hydrocarbons in a clade of cactophilic Drosophila

Background: We characterized variation and chemical composition of epicuticular hydrocarbons (CHCs) in the seven species of the Drosophila buzzatii cluster with gas chromatography/mass spectrometry. Despite the critical role of CHCs in providing resistance to desiccation and involvement in communication, such as courtship behavior, mating, and aggregation, few studies have investigated how CHC profiles evolve within and between species in a phylogenetic context. We analyzed quantitative differences in CHC profiles in populations of the D. buzzatii species cluster in order to assess the concordance of CHC differentiation with species divergence. Results: Thirty-six CHC components were scored in single fly extracts with carbon chain lengths ranging from C(29) to C(39), including methyl-branched alkanes, n alkenes, and alkadienes. Multivariate analysis of variance revealed that CHC amounts were significantly different among all species and canonical discriminant function (CDF) analysis resolved all species into distinct, non-overlapping groups. Significant intraspecific variation was found in different populations of D. serido suggesting that this taxon is comprised of at least two species. We summarized CHC variation using CDF analysis and mapped the first five CHC canonical variates (CVs) onto an independently derived period (per) gene + chromosome inversion + mtDNA COI gene for each sex. We found that the COI sequences were not phylogenetically informative due to introgression between some species, so only per + inversion data were used. Positive phylogenetic signal was observed mainly for CV1 when parsimony methods and the test for serial independence (TFSI) were used. These results changed when no outgroup species were included in the analysis and phylogenetic signal was then observed for female CV3 and/or CV4 and male CV4 and CV5. Finally, removal of divergent populations of D. serido significantly increased the amount of phylogenetic signal as up to four out of five CVs then displayed positive phylogenetic signal. Conclusions: CHCs were conserved among species while quantitative differences in CHC profiles between populations and species were statistically significant. Most CHCs were species-, population-, and sex-specific. Mapping CHCs onto an independently derived phylogeny revealed that a significant portion of CHC variation was explained by species' systematic affinities indicating phylogenetic conservatism in the evolution of these hydrocarbon arrays, presumptive waterproofing compounds and courtship signals as in many other drosophilid species.

Genetic variability of the Amazon River prawn Macrobrachium amazonicum (Decapoda, Caridea, Palaemonidae)

The freshwater prawn Macrobrachium amazonicum is widely distributed in South America, and occupies habitats with a wide range of salinities. Several investigations have revealed the existence of wide intraspecific variability among different populations, although the understanding of this variability is still fragmentary and incomplete. We compared and characterized inland and coastal populations of M. amazonicum from Brazil, using molecular data (16S and COI mtDNA) to describe the degree of variability, structure, and relationships among them. Genetic divergence rates among populations showed variability at the intraspecific level. All the analyses evidenced significant genetic divergence among populations, structuring them in three groups: I-inland waters of the Amazonian Hydrographic Region (HR); II-Parana/Paraguay HR; and III-coastal systems of northern and northeastern Brazil. Phylogenetic reconstructions revealed that the populations form a single monophyletic clade, which supports their characterization as a single species. Clade I was a sister clade of that formed by clades II and III, which were themselves sister clades. Populations from Sertaozinho/Miguelopolis and Avare, introduced into the state of Sao Paulo, may have originated from natural populations in the states of Mato Grosso do Sul and Para, respectively. Geographical isolation probably contributed to the observed variation, and if this isolation continues. M. amazonicum may undergo speciation within its broad geographical distribution. The sequences obtained here can be used as name-tags for population identification, and the DNA barcodes are useful to identify the origin of specimens used in different freshwater-prawn cultures or introduced populations of unknown origin.

Influence of memory in deterministic walks in random media: Analytical calculation within a mean-field approximation

Consider a random medium consisting of N points randomly distributed so that there is no correlation among the distances separating them. This is the random link model, which is the high dimensionality limit (mean-field approximation) for the Euclidean random point structure. In the random link model, at discrete time steps, a walker moves to the nearest point, which has not been visited in the last mu steps (memory), producing a deterministic partially self-avoiding walk (the tourist walk). We have analytically obtained the distribution of the number n of points explored by the walker with memory mu=2, as well as the transient and period joint distribution. This result enables us to explain the abrupt change in the exploratory behavior between the cases mu=1 (memoryless walker, driven by extreme value statistics) and mu=2 (walker with memory, driven by combinatorial statistics). In the mu=1 case, the mean newly visited points in the thermodynamic limit (N >> 1) is just < n >=e=2.72... while in the mu=2 case, the mean number < n > of visited points grows proportionally to N(1/2). Also, this result allows us to establish an equivalence between the random link model with mu=2 and random map (uncorrelated back and forth distances) with mu=0 and the abrupt change between the probabilities for null transient time and subsequent ones.

Endoscopic Stented Ureterocystostomy

Background and Purpose: Fibrotic or neoplastic obstruction of the terminal ureter and ureterovesical junction can preclude internal drainage with a Double-J catheter. Some minimally invasive alternatives are described in the literature to avoid a percutaneous nephrostomy. We present a pure endourologic technique. Patients and Methods: In six patients with an obstructed upper urinary tract, after the introduction of iodine contrast, the ureter was punctured with a needle to introduce a guidewire in the urinary tract under cystoscopic and fluoroscopic control. The alternative path between the bladder and ureter was then dilated up 10F to facilitate the Double-J catheter introduction. Results: All six patients had their obstructed urinary tract drained with a Double-J catheter inserted above the level of obstruction. No complication was verified. Conclusion: Internal urinary tract drainage with a Double-J catheter was accomplished using endourologic principles in six patients, avoiding a percutaneous nephrostomy or other more invasive procedures.

Characterization of Hepatitis B virus (HBV) genotypes in patients from Rondonia, Brazil

Background: Hepatitis B virus (HBV) can be classified into nine genotypes (A-I) defined by sequence divergence of more than 8% based on the complete genome. This study aims to identify the genotypic distribution of HBV in 40 HBsAg-positive patients from Rondonia, Brazil. A fragment of 1306 bp partially comprising surface and polymerase overlapping genes was amplified by PCR. Amplified DNA was purified and sequenced. Amplified DNA was purified and sequenced on an ABI PRISM (R) 377 Automatic Sequencer (Applied Biosystems, Foster City, CA, USA). The obtained sequences were aligned with reference sequences obtained from the GenBank using Clustal X software and then edited with Se-Al software. Phylogenetic analyses were conducted by the Markov Chain Monte Carlo (MCMC) approach using BEAST v.1.5.3. Results: The subgenotypes distribution was A1 (37.1%), D3 (22.8%), F2a (20.0%), D4 (17.1%) and D2 (2.8%). Conclusions: These results for the first HBV genotypic characterization in Rondonia state are consistent with other studies in Brazil, showing the presence of several HBV genotypes that reflects the mixed origin of the population, involving descendants from Native Americans, Europeans, and Africans.

Maternal immunization with ovalbumin prevents neonatal allergy development and up-regulates inhibitory receptor Fc gamma RIIB expression on B cells

Background: Preconception allergen immunization prevents neonatal allergen sensitization in mice by a complex interaction between regulatory cells/factors and antibodies. The present study assessed the influence of maternal immunization with ovalbumin (OVA) on the immune response of 3 day-old and 3 week-old offspring immunized or non-immunized with OVA and evaluated the effect of IgG treatment during fetal development or neonatal period. Results: Maternal immunization with OVA showed increased levels of Fc gamma RIIb expression in splenic B cells of neonates, which were maintained for up to 3 weeks and not affected by additional postnatal OVA immunization. Maternal immunization also exerted a down-modulatory effect on both IL-4 and IFN-gamma-secreting T cells and IL-4 and IL-12-secreting B cells. Furthermore, immunized neonates from immunized mothers showed a marked inhibition of antigen-specifc IgE Ab production and lowered Th2/Th1 cytokine levels, whereas displaying enhanced Fc gamma RIIb expression on B cells. These offspring also showed reduced antigen-specific proliferative response and lowered B cell responsiveness. Moreover, in vitro evaluation revealed an impairment of B cell activation upon engagement of B cell antigen receptor by IgG from OVA-immunized mice. Finally, in vivo IgG transference during pregnancy or breastfeeding revealed that maternal Ab transference was able to increase regulatory cytokines, such as IL-10, in the prenatal stage; yet only the postnatal treatment prevented neonatal sensitization. None of the IgG treatments induced immunological changes in the offspring, as it was observed for those from OVA-immunized mothers. Conclusion: Maternal immunization upregulates the inhibitory Fc gamma RIIb expression on offspring B cells, avoiding skewed Th2 response and development of allergy. These findings contribute to the advancement of prophylactic strategies to prevent allergic diseases in early life.

Near full-length genome analysis of low prevalent human immunodeficiency virus type 1 subclade F1 in Sao Paulo, Brazil

Background: The genetic diversity of the human immunodeficiency virus type 1 (HIV-1) is critical to lay the groundwork for the design of successful drugs or vaccine. In this study we aimed to characterize and define the molecular prevalence of HIV-1 subclade F1 currently circulating in Sao Paulo, Brazil. Methods: A total of 36 samples were selected from 888 adult patients residing in Sao Paulo who had previously been diagnosed in two independent studies in our laboratory as being infected with subclade F1 based on pol subgenomic fragment sequencing. Proviral DNA was amplified from the purified genomic DNA of all 36 blood samples by 5 fragments overlapping PCR followed by direct sequencing. Sequence data were obtained from the 5 fragments of pure subclade F1 and phylogenetic trees were constructed and compared with previously published sequences. Subclades F1 that exhibited mosaic structure with other subtypes were omitted from any further analysis Results: Our methods of fragment amplification and sequencing confirmed that only 5 sequences inferred from pol region as subclade F1 also holds true for the genome as a whole and, thus, estimated the true prevalence at 0.56%. The results also showed a single phylogenetic cluster of the Brazilian subclade F1 along with non-Brazilian South American isolates in both subgenomic and the full-length genomes analysis with an overall intrasubtype nucleotide divergence of 6.9%. The nucleotide differences within the South American and Central African F1 strains, in the C2-C3 env, were 8.5% and 12.3%, respectively. Conclusion: All together, our findings showed a surprisingly low prevalence rate of subclade F1 in Brazil and suggest that these isolates originated in Central Africa and subsequently introduced to South America.

ISOLATION OF Rickettsia bellii FROM Amblyomma ovale AND Amblyomma incisum TICKS FROM SOUTHERN BRAZIL

Objective. To isolate and characterize rickettsiae from the ticks Amblyomma ovale and Amblyomma incisum collected in the state of Sao Paulo. Materials and methods. Adult, free-living A. ovale and A. incisum were collected in an Atlantic rainforest area in the state of Sao Paulo, Brazil. Each tick was tested using the hemolymph assay; samples from positive ticks were placed in shell vials in order to isolate rickettsiae and subsequently grown in Vero cells. Amplification of three rickettsial genes ( gltA, htrA and ompA) was attempted using polymerase chain reaction (PCR) for each isolate obtained. Amplicons were subsequently sequenced. Results. A total of 388 A. incisum and 50 A. ovale were collected. Only one A. incisum and one A. ovale were hemolymph-test positive. Rickettsiae were successfully isolated from these ticks; however establishment in Vero cell culture was successful only for the isolate from A. ovale. Bacterial contamination in the first cell passage of the A. incisum isolate precluded successful isolation of the organism. PCR products were obtained with the gltA and htrA primers for the two isolates, but no product was obtained with the ompA primers. By BLAST analysis, partial gltA and htrA sequences of isolates from A. ovale and A. incisum were similar to the corresponding sequences of R. bellii. Conclusions. This is the first report of R. bellii infecting A. incisum and the first successful isolation from A. ovale.