Diagnóstico molecular da alteração mutagênica nt 230 (del4) no gene MDR1 em cães

P-glycoprotein is an adenosine triphosphate (ATP)-driven drug efflux carrier responsible for transport of xenobiotics and multiple classes of drugs, many usually use in veterinary medicine. Encoded by MDR1 gene, also referred to as ABCB1, located on chromosome 14, is expressed in many tissues with secretory or excretory functions, such as liver, kidney and intestine, where it limits drug absorption from the gut and promotes drug excretion into the bile and urine of their substrates. In 2001, a 4 base pair gene deletion mutation in the canine MDR1 gene was identified as MDR1-1▲, ABCB1-1▲, MDR1 MDR1 nt 230 (del4) and associated with an non-functional Pglycoprotein. The clinical correlation is the (hyper) sensitivity of certain dogs breeds, mostly collies, to a few classes of drugs such as anticancer drugs (doxorubicin, vincristine, vinblastine), immunosuppressants (cyclosporine), antiparasitic drugs (ivermectin, moxidectin), steroids hormones (aldosterone, cortisol, dexamethasone), antimicrobial agents (tetracycline, doxycycline, levofloxacin, ketoconazole, itraconazole), analgesics (morphine, methadone), antidiarrheals (loperamide), antiepileptic agents (phenothiazine), cardiac drugs (digoxin, diltiazem, verapamil, talinolol) and others. Dogs with homozygous MDR1 nt 230 (del4) MDR1 mutations (MDR1 - / -) have a higher predisposition to intoxication with substrates of P-gp than heterozygous (MDR1 + / -) and these are more likely than dogs homozygous nonmutant (MDR1 +/ +). After the identification of nt230 (del4) mutation, several molecular techniques have been developed for identification of mutant animals as a diagnostic method. The importance of molecular diagnosis is, after the identification of mutant animals, establish treatment protocols safe, exclude this animals from reproduction (genetic selection program) and investigating the history of adverse drugs reactions... (Complete abstract click electronic access below)

Functional XPB/RAD25 redundancy in Arabidopsis genome: characterization of AtXPB2 and expression analysis

The xeroderma pigmentosum complementation group B (XPB) protein is involved in both DNA repair and transcription in human cells. It is a component of the transcription factor IIH (TFIIH) and is responsible for DNA helicase activity during nucleotide (nt) excision repair (NER). Its high evolutionary conservation has allowed identification of homologous proteins in different organisms, including plants. In contrast to other organisms, Arabidopsis thaliana harbors a duplication of the XPB orthologue (AtXPB1 and AtXPB2), and the proteins encoded by the duplicated genes are very similar (95% amino acid identity). Complementation assays in yeast rad25 mutant strains suggest the involvement of AtXPB2 in DNA repair, as already shown for AtXPB1, indicating that these proteins may be functionally redundant in the removal of DNA lesions in A. thaliana. Although both genes are expressed in a constitutive manner during the plant life cycle, Northern blot analyses suggest that light modulates the expression level of both XPB copies, and transcript levels increase during early stages of development. Considering the high similarity between AtXPB1 and AtXPB2 and that both of predicted proteins may act in DNA repair, it is possible that this duplication may confer more flexibility and resistance to DNA damaging agents in thale cress. (C) 2004 Elsevier B.V. All rights reserved.

Changes in biomass, chemical composition and nutritive value of Spartina alterniflora due to organic pollution in the Itanhaém River Basin (SP, Brazil)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Adubação nitrogenada para milho com o uso de plantas de cobertura e modos de aplicação de calcário

O sistema plantio direto (SPD) é uma realidade na região dos Cerrados, mas alguns questionamentos persistem nesse tipo de manejo como o modo de realização da calagem e a dose de nitrogênio (N) a ser adotada em cultura comercial, em relação às culturas precedentes. Desse modo, objetivou-se avaliar modos de aplicar o calcário na implantação do SPD e o efeito de culturas de cobertura precedentes sobre a necessidade de adubação nitrogenada da cultura do milho, durante diferentes anos agrícolas. O delineamento experimental utilizado foi de blocos casualizados, em esquema fatorial 5 x 2 (modos de aplicação do calcário x culturas de cobertura) e posterior divisão em três subparcelas, referentes às doses de N (0, 90 e 180 kg ha-1). Foram avaliados quatro modos de aplicação de calcário: incorporado a 0-0,2 m, em out./2001; dose total em superfície aplicada, em out./2001; aplicação de 1/2 da dose, em out./2001, e 1/2, em ago./2002, na superfície; e aplicação de 1/3 da dose recomendada, em mar./2001, + 1/3, em out./2001, e 1/3, em ago./2002, também em superfície. Além de um tratamento testemunha (sem calcário) e duas culturas de cobertura, crotalária e milheto. Os diferentes modos de calagem não alteraram a produtividade de grãos de milho. O cultivo de milho após crotalária apresentou melhor desempenho e menor demanda de adubação nitrogenada, quando comparado ao cultivado após milheto.

Short-term temporal changes of soil carbon losses after tillage described by a first-order decay model

Tillage stimulates soil carbon (C) losses by increasing aeration, changing temperature and moisture conditions, and thus favoring microbial decomposition. In addition, soil aggregate disruption by tillage exposes once protected organic matter to decomposition. We propose a model to explain carbon dioxide (CO2) emission after tillage as a function of the no-till emission plus a correction due to the tillage disturbance. The model assumes that C in the readily decomposable organic matter follows a first-order reaction kinetics equation as: dC(sail)(t)/dt = -kC(soil)(t) and that soil C-CO2 emission is proportional to the C decay rate in soil, where C-soil(t) is the available labile soil C (g m(-2)) at any time (t). Emissions are modeled in terms soil C available to decomposition in the tilled and non-tilled plots, and a relationship is derived between no-till (F-NT) and tilled (F-Gamma) fluxes, which is: F-T = a1F(NT)e(-a2t), where t is time after tillage. Predicted and observed fluxes showed good agreement based on determination coefficient (R-2), index of agreement and model efficiency, with R-2 as high as 0.97. The two parameters included in the model are related to the difference between the decay constant (k factor) of tilled and no-till plots (a(2)) and also to the amount of labile carbon added to the readily decomposable soil organic matter due to tillage (a,). These two parameters were estimated in the model ranging from 1.27 and 2.60 (a(1)) and - 1.52 x 10(-2) and 2.2 x 10(-2) day(-1) (a(2)). The advantage is that temporal variability of tillage-induced emissions can be described by only one analytical function that includes the no-till emission plus an exponential term modulated by tillage and environmentally dependent parameters. (C) 2008 Elsevier B.V. All rights reserved.

Short-term temporal changes of bare soil CO2 fluxes after tillage described by first-order decay models

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The kinetics of donor cell mtDNA in embryonic and somatic donor cell-derived bovine embryos

The mechanisms controlling the outcome of donor cell-derived mitochondrial DNA (mtDNA) in cloned animals remain largely unknown. This research was designed to investigate the kinetics of somatic and embryonic mtDNA in reconstructed bovine embryos during preimplantation development, as well as in cloned animals. The experiment involved two different procedures of embryo reconstruction and their evaluation at five distinct phases of embryo development to measure the proportion of donor cell mtDNA (Bos indicus), as well as the segregation of this mtDNA during cleavage. The ratio of donor cell (B. indicus) to host oocyte (B. taurus) mtDNA (heteroplasmy) from blastomere- (NT-B) and fibroblast- (NT-F) reconstructed embryos was estimated using an allele-specific PCR with fluorochrome-stained specific primers in each sampled blastomere, in whole blastocysts, and in the tissues of a fibroblast-derived newborn clone. NT-B zygotes and blastocysts show similar levels of heteroplasmy (11.0% and 14.0%, respectively), despite a significant decrease at the 9-16 cell stage (5.8%; p < 0.05). Heteroplasmy levels in NT-F reconstructed zygotes, however, increased from an initial low level (4.7%), to 12.9% (p < 0.05) at the 9-16 cell stage. The NT-F blastocysts contained low levels of heteroplasmy (2.2%) and no somatic-derived mtDNA was detected in the gametes or the tissues of the newborn calf cloned. These results suggest that, in contrast to the mtDNA of blastomeres, that of somatic cells either undergoes replication or escapes degradation during cleavage, although it is degraded later after the blastocyst stage or lost during somatic development, as revealed by the lack of donor cell mtDNA at birth.

Demecolcine Effects on Microtubule Kinetics and on Chemically Assisted Enucleation of Bovine Oocytes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Microbial populations and the activity of the soil under agricultural and agricultural-pastoral systems

The effects of agricultural-pastoral and tillage practices on soil microbial populations and activities have not been systematically investigated. The effect of no-tillage (NT), no-tillage agricultural-pastoral integrated systems (NT-I) and conventional tillage (CT) at soil depths of 0-10, 10-20 and 20-30 cm on the microbial populations (bacteria and fungi), biomass-C, potential nitrification, urease and protease activities, total organic matter and total N contents were investigated. The crops used were soybean (in NT, NT-I and CT systems), corn (in NT and NT-I systems) and Tanner grass (Brachiaria sp.) (in NT-I system); a forest system was used as a control. Urease and protease activities, biomass-C and the content of organic matter and total N were higher (p < 0.05) in the forest soil than the other soils. Potential nitrification was significantly higher in the NT-I system in comparison with the other systems. Bacteria numbers were similar in all systems. Fungi counts were similar in the CT and forest, but both were higher than in NT. All of these variables were dependent on the organic matter content and decreased (p < 0.05) from the upper soil layer to the deeper soil layers. These results indicate that the no-tillage agricultural-pasture-integrated systems may be useful for soil conservation.

Co-accumulation of microbial residues and particulate organic matter in the surface layer of a no-till Oxisol under different crops

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fine root isotropy in Eucalyptus grandis plantations. Towards the prediction of root length densities from root counts on trench walls

The objectives of the study were to assess changes in fine root anisotropy and specific root lengths throughout the development of Eucalyptus grandis ( W. Hill ex Maiden) plantations and to establish a predictive model of root length density (RLD) from root intercept counts on trench walls. Fine root densities (<1 mm in diameter) were studied in 6-, 12-, 22-, 28-, 54-, 68- and 72-month-old E. grandis plantations established on deep Ferralsols in southern Brazil. Fine root intercepts were counted on 3 faces of 90-198 soil cubes (1 dm(3) in volume) in each stand and fine root lengths (L) were measured inside 576 soil cubes, sampled between the depths of 10 cm and 290 cm. The number of fine root intercepts was counted on one vertical face perpendicular to the planting row (N(t)), one vertical face parallel to the planting row (N(l)) and one horizontal face (N(h)), for each soil cube sampled. An overall isotropy of fine roots was shown by paired Student's t-tests between the numbers of fine roots intersecting each face of soil cubes at most stand ages and soil depths. Specific root lengths decreased with stand age in the upper soil layers and tended to increase in deep soil layers at the end of the rotation. A linear regression established between N(t) and L for all the soil cubes sampled accounted for 36% of the variability of L. Such a regression computed for mean Nt and L values at each sampling depth and stand age explained only 55% of the variability, as a result of large differences in the relationship between L and Nt depending on stand productivity. The equation RLD=1.89*LAI*N(t), where LAI was the stand leaf area index (m(2) m(-2)) and Nt was expressed as the number of root intercepts per cm(2), made it possible to predict accurately (R(2)=0.84) and without bias the mean RLDs (cm cm(-3)) per depth in each stand, for the whole data set of 576 soil cubes sampled between 2 years of age and the end of the rotation.

Nitrogen Supply for Cover Crops and Effects on Peanut Grown in Succession under a No-Till System

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Least limiting water range in soil under crop rotations and chiseling

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)