Degradação in situ da matéria seca e da fração fibrosa do capim marandu obtido por diferentes métodos de amostragem, no período seco do ano

Este trabalho foi realizado na Faculdade de Ciências Agrárias Veterinárias - Unesp - Jaboticabal e foi conduzido, com o objetivo de avaliar a influência do método de amostragem do pasto sobre a degradabilidade in situ da matéria seca e da fração fibrosa de capim Marandu, colhido no período seco dos anos de 2003 e 2005. O experimento foi instalado em delineamento de blocos casualizados com parcela subdividida, com três repetições, representadas pelos piquetes amostrados. Nas parcelas, foram avaliados cinco métodos de amostragens de forragem (método do quadrado metálico; método de avaliação através de extrusa de bovino da raça Nelore; método de avaliação por meio de extrusa de bovino Cruzado (Red Angus x Nelore); método de avaliação por meio do pastejo, simulando bovino da raça Nelore; método de avaliação por meio do pastejo, simulando bovino Cruzado (Red Angus x Nelore) e as subparcelas foram constituídas pelos anos de amostragem, 2003 e 2005. Foram determinadas as frações da cinética ruminal: solúvel A; insolúvel potencialmente degradável B; taxa de degradação Kd; degradação potencial (DP) e fração não degradável C da MS e degradação potencial (DP), fração não degradável C e taxa de degradação Kd da FDN e da FDA. de acordo com os resultados obtidos, observou-se que o método do quadrado metálico subestimou as características da degradação do capim. Os métodos do pastejo simulado se assemelharam muito ao das extrusas, no entanto, a prática do simulador é que assegurou a amostragem eficiente, conforme foi constatado pelos dados obtidos no ano de 2003 e 2005.

Degradabilidade in situ e observações microscópicas de volumosos em bovinos suplementados com enzimas fibrolíticas exógenas

Avaliou-se o efeito de enzimas fibrolíticas (celulase e xilanase) sobre a degradabilidade in situ da MS, PB, FDN, FDA e hemicelulose do feno de Tifton-85 (Cynodon spp.) cortado aos 30 e 90 dias e do bagaço de cana, utilizando-se seis bovinos com cânula no rúmen. As enzimas foram extraídas dos fungos Aspergillus niger e Trichoderma longibrachiatum e fornecidas, na quantidade de 0,75 g/kgMS.dia, por meio da cânula ruminal. Os tempos de incubação ruminal foram de 0, 3, 6, 12, 24, 48, 72 e 96 horas. Os resíduos de incubação foram avaliados por meio de microscopia eletrônica de varredura (MEV). O efeito da adição de enzimas sobre a degradação da MS e PB variou em função do volumoso estudado. A degradabilidade efetiva da MS do feno Tifton cortado aos 30 e 90 dias e do bagaço de cana sem a adição de enzimas foi de 61,85; 42,35 e 28,22%, respectivamente, e de 63,51; 40,64 e 31,43%, respectivamente, com a adição das enzimas. Não houve efeito das enzimas sobre a degradação da fibra. As observações ao MEV indicaram aumento da colonização bacteriana sobre a parede celular com a suplementação enzimática. A adição de enzimas fibrolíticas na dieta de ruminantes apresentou efeito pouco expressivo sobre os parâmetros de degradação ruminal dos volumosos estudados.

Efeito do emurchecimento e da adição de fubá na degradabilidade in situ da silagem de alfafa (Medicago sativa L.)

O objetivo deste ensaio foi avaliar o efeito do emurchecimento e da adição de fubá durante o processo de ensilagem da alfafa sobre a degradabilidade in situ da matéria seca (MS), da fibra em detergente neutro (FDN) e da proteína bruta (PB). Foram utilizados três bovinos adultos mestiços fistulados no rúmen, nos quais foram incubados seis tipos de silagens (1-alfafa fresca; 2-alfafa fresca +5% de fubá; 3-alfafa fresca +10% de fubá; 4-alfafa emurchecida; 5-alfafa emurchecida + 5% de fubá ; 6-alfafa emurchecida + 10% de fubá) durante três tempos de incubação (6, 24, 96 horas). O delineamento experimental foi em parcelas subdivididas, com tratamentos em parcelas, tempos em subparcelas e animais como blocos. A adição de fubá à silagem aumentou (P<0,05) a degradação da MS e FDN com o tempo de incubação. O emurchecimento proporcionou diminuição (P<0,05) na degradabilidade dos nutrientes, entretanto esse efeito foi ligeiramente contornado pela adição de 10% de fubá. Os tratamentos com a adição de fubá e o emurchecimento proporcionaram maior taxa de degradabilidade da MS de fração b, entretanto o emurchecimento diminuiu o potencial de degradação a, enquanto que a adição de níveis de fubá não alterou a degradabilidade da MS dessa fração. A degradação da fração b da FDN aumentou com a adição de fubá e diminuiu com o emurchecimento, entretanto houve aumento dessa fração quando se adicionou fubá à forragem emurchecida. A degradação da fração a da PB aumentou com a adição de fubá e diminuiu com o emurchecimento; para a fração b ocorreu o inverso. Concluiu-se que a prática do emurchecimento, por reduzir a fração solúvel da forragem, diminui a degradabilidade dos nutrientes da silagem de alfafa, que no entanto pode ser melhorada com a adição de fubá.

Degradabilidade ruminal in situ da matéria seca e da fibra em detergente neutro em silagens de híbridos de sorgo colhidos em diferentes épocas

O objetivo deste trabalho foi o de avaliar o efeito das silagens de três híbridos de sorgo: granífero (Conti-Silo, de porte baixo), duplo propósito (Conti-Silo-03, de porte médio) e forrageiro (547-F, de porte alto) e de três épocas distintas (aos 105, 112 e 119 dias após a semeadura) sobre a degradabilidade in situ da matéria seca (MS) e da fibra em detergente neutro (FDN). Foram utilizados três bovinos adultos mestiços fistulados no rúmem, distribuídos em um delineamento experimental em parcelas subdivididas, com três tempos de incubação (6, 24, 96 horas). Houve diferença (P<0,05) nas frações de desaparecimento da MS e de FDN após 6, 24 e 96 horas de incubação ruminal entre as diferentes silagens. Quanto à época de colheita, a diferença na degradação de MS foi observada após 96 horas de incubação, destacando-se a variedade de duplo propósito, superior nos três diferentes cortes. Com a maturação, observou-se tendência de aumento na fração degradável da MS, porém esse efeito foi menos evidente nas silagens de sorgo forrageiro. Para a FDN não houve diferença (P<0,05) entre silagens em nenhum dos tempos de incubação.

Influência do nitrogênio degradável no rúmen sobre a degradabilidade in situ, os parâmetros ruminais e a eficiência de síntese microbiana em novilhos alimentados com cana-de-açúcar

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Fission track and U-Pb in situ dating applied to detrital zircon from the Vale do Rio do Peixe Formation, Bauru Group, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Zircon fission track and U-Th-Pb in situ dating of Rio Parana Formation, Parana Basin, Brazil

Ages of zircon from sedimentary samples of Rio Parana Formation, belonging of Bauru Group, north of Parana Basin, Brazil, has been determined by zircon Fission Track and U-Th-Pb in situ dating methods. The obtained ages are from same zircon grain that provided information on the source areas for the sediments and the morphotectonic events.

Impact of the in situ formed salivary pellicle on enamel and dentine erosion induced by different acids

Objective. To investigate and compare the protective impact of the in situ formed salivary pellicle on enamel and dentine erosion caused by different acids at pH 2.6. Methods. Bovine enamel and dentine samples were exposed for 120 min in the oral cavity of 10 healthy volunteers. Subsequently, enamel and dentine pellicle-covered specimens were extraorally immersed in 1 ml hydrochloric, citric or phosphoric acid (pH 2.6, 60 s, each acid n=30 samples). Pellicle-free samples (each acid n=10) served as controls. Calcium release into the acid was determined by atomic absorption spectroscopy. The data were analysed by two-way ANOVA and Tukey's test (alpha=0.05). Results. Pellicle-covered samples showed significantly less calcium loss compared to pellicle-free samples in all acid groups. The mean (SD) pellicle protection (% reduction of calcium loss) was significantly better for enamel samples [60.9 (5.3)] than for dentine samples [30.5 (5.0)], but revealed no differences among the acids. Conclusion. The efficacy of the in situ pellicle in reducing erosion was 2-fold better for enamel than for dentine. Protection of the pellicle was not influenced by the kind of acid when enamel and dentine erosion was performed at pH 2.6.

Efeito de diferentes níveis de bicarbonato de sódio em dietas com bagaço de cana-de-açúcar auto-hidrolisado sobre a degradação in situ do milho e farelo de algodão

The objective of this work was to evaluate the buffering effect of sodium bicarbonate (NaHCO3), used in the levels of: 0, 0.7, 1.4 and 2.1% of dry matter, on the in situ degradation of corn and cottonseed meal. A diet with 60% autohidrolised sugar bagasse (BAH) and 40% of concentrate was used, plus urea, minerals and limestone. The rations was calculated to allow 300g of daily gain. After 20 days of adaptation to the treatment (levels of NaHCO3), 5g of each feed was incubated in the rumen of four bovines for 3, 6, 12, 24 and 48 hours, using naylon bag with size of 7,5 x 17,5 cm with pores of 36 micras. A randomized blocks design with four treatment (levels of NaHCO3) were used. The buffer affected the in situ dry matter degradation, whose means were 49.68; 63,10; 67,71; and 60,85% and 25.89; 30.88; 33.48 and; 31.02% for the corn and cottonseed meal, respectively. The level of 1.4% of NaHCO3 provided the highest value of degradability, which did not differ from the 0.7% and 2.1% levels, for the corn. The degradability of protein was not affected by the treatments.

In situ application of a cellulose bag and an ion exchanger for differentiation of labile and inert metal species in aquatic systems

This work involved the development and application of a new analytical procedure for in-situ characterization of the lability of metal species in aquatic systems by using a system equipped with a diffusion membrane and cellulose organomodified with p-aminobenzoic acid groups (DM-Cell-PAB). To this end, the DM-Cell-PAB system was prepared by adding cellulose organomodified with p-aminobenzoic acid groups (Cell-PAB) to pre-purified cellulose bags. After the DM-Cell-PAB system was sealed, it was examined in the laboratory. The in-situ application involved immersing the DM-Cell-PAB system in two different rivers, enabling us to study the relative lability of metal species (Cu, Cd, Fe, Mn, and Ni) as a function of time and quantity of exchanger. The procedure is simple and opens up a new perspective for understanding environmental phenomena relating to the complexation, transport, stability, and lability of metal species in aquatic systems rich in organic matter.

Approach combining on-line metal exchange and tangential-flow ultrafiltration for in-situ characterization of metal species in humic hydrocolloids

This paper deals with the development and optimization of an analytical procedure using ultrafiltration and a flow-injection system, and its application in in-situ experiments to characterize the lability and availability of metal species in humic-rich hydrocolloids. The on-line system consists of a tangential flow ultrafiltration device equipped with a 3-kDa filtration membrane. The concentration of free ions in the filtrate was determined by atomic-absorption spectrometry, assuming that metals not complexed by aquatic humic substances (AHS) were separated from the complexed species (M-AHS) retained by the membrane. For optimization, exchange experiments using Cu(II) solutions and AHS solutions doped with the metal ions Ni(II), Mn(II), Fe(III), Cd (II), and Zn(II) were carried out to characterize the stability of the metal-AHS complexes. The new procedure was then applied in-situ at a tributary of the Ribeira do Iguape river (Iguape, São Paulo State, Brazil) and evaluated using the ions Fe(III) and Mn(II), which are considered to be essential constituents of aquatic systems. From the exchange between metal-natural organic matter (M-NOM) and the Cu(II) ions it was concluded that Cu(II) concentrations > 485 mu g L(-1) were necessary to obtain maximum exchange of the complexes Mn-NOM and Fe-NOM, corresponding to 100% Mn and 8% Fe. Moreover, the new analytical procedure is simple and opens up new perspectives for understanding the complexation, transport, stability, and lability of metal species in humic-rich aquatic environments.

Efeito de Diferentes Níveis de bicarbonato de sódio sobre a degradação in situ do bagaço de cana-de-açucar auto-hidrolisado

The objective of this work was to evaluate the buffering effects, using sodium bicarbonate (NaHCO3) at the levels: 0; 0.7; 1.4; and 2.1% of dry matter, on the in situ degradation of autohidrolised sugar-cane bagasse. A diet was used with 60% autohidrolised sugar bagasse (AHB) and 40% of concentrate, plus urea, minerals and limestone. The rations was calculated to allow 300g of daily gain. After 20 days adaptation to the treatment (level of NaHCO3), 5 g of AWE was incubated on rumen of four bovines for 3, 6, 12, 24 and 48 hours, using naylon bag measuring 7,5 x 17,5 cm with pores of 36 micras. It was used a randomized blocks design with four treatments (NaHCO3) and four replications(animals). For the calculations of the protein degradation, it was considered the soluble fraction in water plus the degraded fraction in the same proportion as for the neutral detergent fiber (NDF). It was observed that the buffer did not affect the degradability in situ of AHB, whose averages in 48 hours of incubation for dry matter, organic matter, crude protein and neutral detergente fiber (NDF) were 34.83; 36.90; 55.40; and 25.56%, respectively.

Borderline leprosy: in situ and cytokine profile in supernatant of mononuclear of cell culture

In order to contribute to a better understanding of cytokine participation in borderline leprosy, in the present study we determined - by in vitro and in situ examinations - the production of these cytokine mediation in non-treated borderline tuberculoid (BT) patients and borderline lepromatous (BL) patients. Seven non-treated BT patients, 12 non-treated BL patients, besides 19 healthy individuals (control group), were evaluated. Peripheral blood mononuclear cells (PBMC) were stimulated or not with specific-M. leprae stimulus (whole and sonicated M. leprae antigens) and a non-specific stimulus. After 48 hours, supernatant was collected for TNF-alpha, IFN-gamma, IL-10 and TGF-beta1 cytokine determination by ELISA. Biopsies from cutaneous lesions were submitted to histological analysis and hematoxylin-eosin and Fite-Faraco stainings; the sections then underwent iNOS, IL-10 and TGF-beta1 in situ detection by immunohistochemistry. Cytokine quantification in PBMC supernatants from patients showed that BT patients produced higher levels of IFN-gamma. Compared to healthy individuals, both borderline patient groups produced lower levels of TGF-beta1 while BL patients generated lower IL-10 levels. The in situ iNOS expression was higher in BT patients compared to BL individuals. on the order hand, TGF-beta1 cytokine revealed a higher proportion of immunostained cells in BL patients. There was no significant difference in IL-10 level between BT and BL patients. Regarding cutaneous lesions, in BL patients there was a negative correlation between TGF-beta1 tissue expression and IL-10. Independently of the clinical form, we observed a positive correlation between TGF-beta1 and bacterial index as well as a negative correlation between the TGF-beta1 tissue expression and iNOS. The results even showed a positive correlation between iNOS tissue expression and production of IFN-gamma by PBMC stimulated with M. leprae antigens. Taken together, the histopathological and immunological observations reinforce the notion of immunological instability in borderline leprosy patients and indicating the participation of mixed cytokines profiles in these individuals, specifically a Th1 profile in BT patients and Th2 profile in BL patients, with a possible participation of T-regulatory lymphocytes.

Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.