Modelo teórico e experimental da reciclagem do Carbono-13 em tecidos de mamíferos e aves

A diferença entre fontes alimentares da ordem de 14 , originárias de plantas com ciclos fotossintéticos Carbono-3 (C3) e Carbono-4 (C4) e seus subprodutos, abre novas perspectivas para o estudo do metabolismo do carbono em aves e animais de pequeno porte. Os autores propõem um modelo teórico e experimental capaz de exprimir os resultados de enriquecimento relativo, delta per mil (delta ) da razão 13C/12C versus tempo em diferentes tecidos. Utilizou-se a equação y(t) = (y0 -- q/k) e-kt + q/k onde, y(t) é a concentração isotópica no tempo desejado, y0 a concentração isotópica inicial existente no tecido, k é uma constante de troca isotópica com unidade 1/tempo, t é unidade de tempo e q é a taxa de entrada de metabólitos que contém carbono, com valores de delta /tempo. Para fígado de galinhas que tiveram a ração de ciclo fotossintético C4 substituída por dieta C3 obteve-se a equação delta13C = -24,74 + 12,37 e-0.237(nT), com meia-vida (T) de 2,9 dias. O patamar de equilíbrio de substituição do carbono foi alcançado em --24,48 , de modo que praticamente 98,4% do conteúdo isotópico do C4 no fígado foi substituído por C3 após 5,6 meias-vidas. O modelo foi adequado para determinar o tempo de reciclagem total ou parcial da concentração de carbono nos tecidos em estudo.

Qualidade e valor nutritivo de silagens de capim-marandu (B. brizantha cv. Marandu) produzidas com aditivos ou forragem emurchecida

Avaliaram-se a qualidade e o valor nutritivo de silagens de capim-marandu produzidas com polpa cítrica (PC) peletizada, com aditivo enzimático-bacteriano (ABE) ou com forragem emuchercida (E): T1 - forragem sem tratamento (controle); T2 - 10% de polpa cítrica peletizada (PC); T3 - aditivo enzimático bacteriano (AEB - SIL-ALL-C4 da Alltech), à base de 5 g/L de água/t de forragem; T4 - forragem picada grosseiramente emurchecida por 4 horas ao sol. O capim foi colhido aos 60 dias de rebrota (24% MS). Para avaliar o consumo e a digestibilidade das silagens, foram utilizados 16 novilhos mestiços com 200 kg de PV, que receberam, além das silagens, 1,0 kg de concentrado/animal/dia. O AEB não afetou a composição da silagem, mas a PC e o E aumentaram o teor de MS de 24% para 31 e 48%, respectivamente, e reduziram o pH e os teores de N-NH3 das silagens, que foram, respectivamente, de 4,17 e 4,58 e 6,78 e 7,99% NT. A PC diminuiu os teores de FDN e FDA em 12 e 4 unidades percentuais. O consumo de MS da silagem produzida com capim emurchecido foi superior (111,8 g MS/PV0,75) ao das silagens controle ou com AEB, mas não diferiu do obtido para a silagem com PC, que também não diferiu das demais. Os tratamentos não afetaram a digestibilidade, cujas médias para MS, PB, CT, FDN, FDA e NDT foram 67,0; 65,4; 68,8; 63,0; 62,5; e 65,6%, respectivamente. A PC e o E reduziram a proteólise e estimularam o consumo. A PC, o E e o AEB não melhoraram o valor nutritivo da silagem de capim-marandu colhido com 24% MS.

Aplicação dos isótopos estáveis em aqüicultura

O produto mais importante do conhecimento é a ignorância. A situação sugere a imagem de alguém que vive numa ilha de conhecimento cercado por um mar de ignorância. Quando a área dessa ilha aumenta pelo progresso da ciência, também aumenta o litoral, - a fronteira com o desconhecido. Esta expansão é o fruto mais valioso do conhecimento (David Gross, Nobel em Física, 2004). O uso dos isótopos estáveis dos bioelementos (CHON'S) presentes na Atmosfera, Hidrosfera, Litosfera e Biosfera enquadra-se neste contexto, surgindo como ferramenta de alto potencial em pesquisas nas diversas áreas das Ciências da Vida. Com organismos aquáticos, a aplicação dos isótopos estáveis do carbono (δ 13C) e do nitrogênio (δ 15N) tem ocorrido em diversas áreas. em larvas de pacu Piaractus mesopotamicus, a técnica evidenciou a escolha voluntária entre organismos vivos e dieta seca pelos animais, desde as primeiras alimentações até o final da larvicultura. Na carcinicultura, o crescimento do Penaeus vannamei resultou do aproveitamento dos alimentos oferecidos e da produtividade primária do lago (biota). Por meio do balanço de massa e isotópico foi possível determinar a contribuição da biota, da ordem de 44 a 86% do carbono assimilado pelos animais durante o crescimento. Outro exemplo é com o caranguejo de mangue Sesarma rectum, em que sua preferência alimentar por Spartina alterniflora (C4) versus Rhizophora mangle (C3) ou Hibiscus tiliaceus (C3) foi evidenciada pelo modelo isotópico de duas fontes alimentares e os isótopos estáveis do carbono (δ13C). A técnica dos isótopos estáveis associada ao conhecimento dos conceitos básicos da variabilidade isotópica natural e ao uso criterioso das razões 13C/12C; ²H/¹H; 18O/16O; 15N/14N e 36S/34S revela-se como um avanço na fronteira dos conhecimentos, nas diversas reentrâncias do litoral da aqüicultura.

Avaliação do Metabolismo Nutricional em Poedeiras pela Técnica dos Isótopos Estáveis do Ccarbono (13C/12C)

Os isótopos estáveis do carbono que eram utilizados em estudos ecológicos e paleoecológicos apresentaram um incremento nos últimos trinta anos, na utilização em estudos dietéticos em animais. Entretanto, existem poucas informações sobre o padrão metabólico e sobre as taxas de turnover do 13C em aves. O presente experimento estabeleceu curvas de substituição e taxas de movimentação do 13C no ovo e no fígado de aves de postura adultas, pela substituição da ração comercial por dietas compostas de grãos dos ciclos fotossintéticos C3 e C4, durante 50 dias. A diferença no conteúdo isotópico do delta per mil do carbono-13 (delta 13C) entre as duas dietas foi de 16,13 . A taxa de substituição do 13C das dietas, nos tecidos, adequou-se num modelo exponencial, descrevendo o turnover do carbono nos tecidos analisados. As taxas de movimentação do 13C, nas aves alimentadas com dieta baseada em grãos C3, foi maior no fígado em relação ao ovo, com valores para a meia-vida de 2,9 e 3,7 dias, respectivamente. As aves que receberam ração com grãos C4 apresentaram uma taxa de turnover no ovo superior àquela obtida para o fígado, com meia-vida de 4,0 e 5,3 dias, respectivamente. Os valores do delta 13C observados para ovo e fígado diferiram em aproximadamente 2 daqueles referentes às dietas.

Precipitation chemistry in semi-arid areas of Southern Africa: A case study of a rural and an industrial site

Experimental data on the precipitation chemistry in the semi-arid savanna of South Africa is presented in this paper. A total of 901 rainwater samples were collected with automatic wet-only samplers at a rural site, Louis Trichardt, and at an industrial site, Amersfoort, from July 1986 to June 1999. The chemical composition of precipitation was analysed for seven inorganic and two organic ions, using ion chromatography. The most abundant ion was SO(4)(2-) and a large proportion of the precipitation is acidic, with 98% of samples at Amersfoort and 94% at Louis Trichardt having a pH below 5.6 ( average pH of 4.4 and 4.9, respectively). This acidity results from a mixture of mineral and organic acids, with mineral acids being the primary contributors to the precipitation acidity in Amersfoort, while at Louis Trichardt, organic and mineral acids contribute equal amounts of acidity. It was found that the composition of rainwater is controlled by five sources: marine, terrigenous, nitrogenous, biomass burning and anthropogenic sources. The relative contributions of these sources at the two sites were calculated. Anthropogenic sources dominate at Amersfoort and biomass burning at Louis Trichardt. Most ions exhibit a seasonal pattern at Louis Trichardt, with the highest concentrations occurring during the austral spring as a result of agricultural activities and biomass combustion, while at Amersfoort it is less pronounced due to the dominance of relatively constant industrial emissions. The results are compared to observations from other African regions.

Morphological changes in the salivary glands of Amblyomma cajennense females (Acari : Ixodidae) in different feeding stages on rabbits at first infestation

This study examined salivary glands of unfed, partially engorged, and engorged females of the tick Amblyomma cajennense on rabbits at first infestation using histological and histochemical techniques. In type I acini, no significant changes were observed among the three feeding conditions. In type II acini of unfed females, c1, c2, and c4 cells were described for the first time in this species. In a comparison among the three feeding conditions, an increase in this acinus was observed, due to the increase in secretion in c1, c2, and c4 cells and the appearance of c3 cells. In engorged females, some cells were still active. Type III acini presented cells d, e, and f containing secretion in unfed females. In partially engorged females, these cells were devoid of secretion. In engorged females, type III acini exhibited a reduced lumen. After engorgement, all acini underwent a degenerative process, as observed in females after two to five days post-engorgement.

Alterações imunitárias na cirrose hepática alcoólica+.

The alcoholic liver cirrhosis usually causes overall immunological changes which might be attributed to either the hepatic disease itself, to ethanol action and/or to malnourishment of the patient. These immune abnormalities comprise both cellular and humoral immunity, consisting of increased immunoglobulin levels, depressed late-skin response to antigens, lowered proliferative response of lymphocytes to mitogens, lower plasma levels of complement proteins (C3 and C4) and by either lower (IL2 and gamma IF) or increased (IL1, TNF, IL6 and IL8) cytokine levels. Parallel to the systemic immune suppression found in most patients, there is also a concomitant local, genetically based, immune stimulation at the liver level which leads to hepatic self-aggression. The systemic immune-suppression could be responsible for periodical infections or neoplasia found in these patients. The possible factors for the immune exhaustion are: a) lower hepatic clearance of toxins and/or bacteria; b) lower hepatic synthesis of complement components; c) cytokines (IL2 and gamma IF) deficiencies, and d) deficiencies of nutrients related to the antioxidant and/or immune defense mechanisms. The immune stimulation of the liver self aggression is characterized by the preferential migration of cytotoxic T cell and neutrophils to the liver, following stimulatory factors such as Mallory bodies, acetaldehyde and/or antibodies. Moreover, the local increase of cytokines (IL1, TNF, IL6 and IL8) levels would be liable for the local phagocyte chemotaxy (IL8) or part of liver injury (TNF) eased by the lower antioxidant defense of the cirrhotic liver.(ABSTRACT TRUNCATED AT 250 WORDS)

The dietary regime of detritivorous fish from the River Jacaré Pepira, Brazil

The diet of detritivorous fish was analysed at four distinct habitats in a tropical river during the wet and dry seasons. Two methodologies were used to determine the diet of the fish: (1) the direct analysis of the stomach contents; and (2) the use of stable carbon isotopes. Both methods indicated food partitioning among the fish. The stomach contents showed what the fish had ingested when captured and the 13C isotope indicated what it had assimilated over a long period of time. The 13C isotope suggested the existence of sectional differences in the feeding of the detritivorous fish along the river. Plants employing the C4 photosynthesis pathways seem to have little influence on the diet of these fish. This method proved to be more appropriate for the detection of the main food sources of the captured fish.

Tracing metabolic routes of feed ingredients in tissues of broiler chickens using stable isotopes

The present study aimed to quantify the proportion of 13C from energy and protein feed ingredients that follow the metabolic routing of the liver and muscle in broiler chickens. A stable isotope of carbon technique was used that is based on the isotopic discrimination that occurs in the plants during the photosynthesis process. One-day-old male chicks were subjected to treatments based on free choice of energy and protein sources. Rice bran (R) and soybean meal (S), C3 plants, have higher isotopic ratios than corn (C), a C4 plant, and corn gluten meal (G). Choices were R+S, C+G, R+G, C+S, or R+C+G+S. A complete feed (CF) was a sixth treatment. Feed intake and BW were measured at 30 d of age, when liver and breast muscle were collected for isotopic analysis. Treatments affected the amount of feed intake and the choices of energy or protein sources. Complete feed had the largest intake, differing from the other treatments that had free-choice feeding. Final BW was a direct reflection of consumption by these birds in all treatments. The isotopic results indicated that the 13C/12C ratio was generally higher in breast muscle than in liver, probably because of higher protein content. Moreover, in the liver, the proportion of 13C retained from the energy ingredient was greater than the proportion from the protein ingredient. That is in contrast to muscle, where the proportion of 13C retained from the protein ingredient was greater than from the energy ingredient that was self-selected.

Carbon isotope composition and leaf anatomy as a tool to characterize the photosynthetic mechanism of Artemisia annua L.

Leaves of Artemisia annua L. are a plentiful source of artemisinin, a drug with proven effectiveness against malaria. The aim of this study was to classify the photosynthetic mechanism of A. annua through studies of the carbon isotope composition (δ 13C) and the leaf anatomy. A. annua presented a δ 13C value of - 31.76 ± 0.07, which characterizes the plants as a typical species of the C3 photosynthethic mechanism, considering that the average δ 13C values for C3 and C4 species are -28 and -14, respectively. The leaf anatomy studies were consistent with the δ 13C results, where, in spite of the existence of parenchymatic cells forming a sheath surrounding the vascular tissue, the cells do not contain chloroplasts or starch. This characteristic is clearly different from that of the Kranz anatomy found in C4 species.

Carbon isotope composition as a tool to control the quality of herbs and medicinal plants

Isotope screening is a simple test for determining the photosynthetic pathway used by plants. The scope of this work was to classify the photosynthetic type of some herbs and medicinal plants through studies of the carbon isotope composition (δ13C). Also, we propose the use of carbon isotope composition as a tool to control the quality of herbs and medicinal plants. For studies of δ13C, δ 13C‰ = [R (sample)/R (standard) - 1] × 10-3, dry leaves powdered in cryogenic mill were analyzed in a mass spectrometer coupled with an elemental analyzer for determining the ratio R = 13CO2/12CO2. In investigation of δ13C of 55 species, 23 botanical families, and 44 species possessed a C3 photosynthetic type. Six species found among the botanical families Euphorbiaceae and Poaceae were C4 plants, and 5 species found among the botanical families Agavaceae, Euphorbiaceae, and Liliaceae possessed CAM-type photosynthesis. Carbon isotope composition of plants can be used as quality control of herbs and medicinal plants, allowing the identification of frauds or contaminations. Also, the information about the photosynthetic type found for these plants can help in introducing and cultivating exotic and wild herbs and medicinal plants.

Carbon isotope composition as a tool for quality control of herbs, spices and medicinal plants

Isotope screening is a simple and cheap test for determining the photosynthetic pathway used by plants. The scope of this work was to classify the photosynthetic type of Melissa officinalis L. and Cymbopogon citratus [DC.] Stapf, through studies of the carbon isotope composition (δ13 C), and we are proposing the use of carbon isotope composition results as a tool to control the quality of medicinal plants. For studies of δ 13C (13 C% = [R (sample)/R (standard) - 1] × 10 -3), dried, powdered leaves were analyzed in a mass spectrometer coupled with an elemental analyzer for determining the ratio R (R = 13CO2/12CO2). As results, M. officinalis presented a C3 photosynthetic type, and C. citratus presented a C4 photosynthetic type. The carbon isotope composition from this study can be used as quality control of M. officinalis adulterants.

The Mycobacterium tuberculosis Rv2540c DNA sequence encodes a bifunctional chorismate synthase

Background. The emergence of multi- and extensively-drug resistant Mycobacterium tuberculosis strains has created an urgent need for new agents to treat tuberculosis (TB). The enzymes of shikimate pathway are attractive targets to the development of antitubercular agents because it is essential for M. tuberculosis and is absent from humans. Chorismate synthase (CS) is the seventh enzyme of this route and catalyzes the NADH- and FMN-dependent synthesis of chorismate, a precursor of aromatic amino acids, naphthoquinones, menaquinones, and mycobactins. Although the M. tuberculosis Rv2540c (aroF) sequence has been annotated to encode a chorismate synthase, there has been no report on its correct assignment and functional characterization of its protein product. Results. In the present work, we describe DNA amplification of aroF-encoded CS from M. tuberculosis (MtCS), molecular cloning, protein expression, and purification to homogeneity. N-terminal amino acid sequencing, mass spectrometry and gel filtration chromatography were employed to determine identity, subunit molecular weight and oligomeric state in solution of homogeneous recombinant MtCS. The bifunctionality of MtCS was determined by measurements of both chorismate synthase and NADH:FMN oxidoreductase activities. The flavin reductase activity was characterized, showing the existence of a complex between FMN ox and MtCS. FMNox and NADH equilibrium binding was measured. Primary deuterium, solvent and multiple kinetic isotope effects are described and suggest distinct steps for hydride and proton transfers, with the former being more rate-limiting. Conclusion. This is the first report showing that a bacterial CS is bifunctional. Primary deuterium kinetic isotope effects show that C4-proS hydrogen is being transferred during the reduction of FMNox by NADH and that hydride transfer contributes significantly to the rate-limiting step of FMN reduction reaction. Solvent kinetic isotope effects and proton inventory results indicate that proton transfer from solvent partially limits the rate of FMN reduction and that a single proton transfer gives rise to the observed solvent isotope effect. Multiple isotope effects suggest a stepwise mechanism for the reduction of FMNox. The results on enzyme kinetics described here provide evidence for the mode of action of MtCS and should thus pave the way for the rational design of antitubercular agents. © 2008 Ely et al; licensee BioMed Central Ltd.

Aprendizado de máquina simbólico e técnicas fractais para caracterizar rejeição em biópsia miocárdica

This work combines symbolic machine learning and multiscale fractal techniques to generate models that characterize cellular rejection in myocardial biopsies and that can base a diagnosis support system. The models express the knowledge by the features threshold, fractal dimension, lacunarity, number of clusters, spatial percolation and percolation probability, all obtained with myocardial biopsies processing. Models were evaluated and the most significant was the one generated by the C4.5 algorithm for the features spatial percolation and number of clusters. The result is relevant and contributes to the specialized literature since it determines a standard diagnosis protocol. © 2013 Springer.

Impact of the length of vitamin D deficiency on cardiac remodeling.

This study was aimed to evaluate the influence of vitamin D (VD) deficiency on cardiac metabolism, morphology, and function. Thus, we investigated the relationship of these changes with the length of the nutrient restriction. Male weanling Wistar rats were allocated into 4 groups: C2 (n=24), animals were fed an AIN-93G diet with 1000 IU VD/kg of chow and were kept under fluorescent light for 2 months; D2 (n=22), animals were fed a VD-deficient AIN-93G diet and were kept under incandescent light for 2 months; C4 (n=21) animals were kept in the same conditions of C2 for 4 months; and D4 (n=23) animals were kept in the same conditions of D2 for 4 months. Biochemical analyses showed lower β-hydroxyacyl coenzyme-A dehydrogenase activity and higher lactate dehydrogenase activity in VD-deficient animals. Furthermore, VD deficiency was related to increased cytokines release, oxidative stress, apoptosis, and fibrosis. Echocardiographic data showed left ventricular hypertrophy and lower fractional shortening and ejection fraction in VD-deficient animals. Difference became evident in the lactate dehydrogenase activity, left ventricular weight, right ventricle weight, and left ventricular mass after 4 months of VD deficiency. Our data indicate that VD deficiency is associated with energetic metabolic changes, cardiac inflammation, oxidative stress, fibrosis and apoptosis, cardiac hypertrophy, left chambers alterations, and systolic dysfunction. Furthermore, length of the restriction influenced these cardiac changes.