Vascularização arterial dos lobos torácicos do timo em fetos de suínos da Linhagem C40

As origens e distribuições das artérias que vascularizaram os lobos torácicos do timo foram estudadas em fetos de 30 suínos da linhagem C40, sendo 12 machos e 18 fêmeas. Os exemplares tiveram o sistema arterial preenchido com solução aquosa a 50% de Neoprene Látex corado e, em seguida, foram submetidos à fixação em solução aquosa a 10% de formaldeído. Os lobos torácicos do timo foram vascularizados por ramos diretos das artérias torácica interna direita (63,33%) e esquerda (53,33%), subclávia esquerda (3,33%), vertebral esquerda (3,33%), cervical superficial direita (3,33%) e esquerda (3,33%), carótida comum esquerda (3,33%), coronária direita (3,33%) e pelos troncos braquiocefálico (33,33%) e costocervical (3,33%). Observaram-se ainda os ramos indiretos das artérias torácica interna direita (70%) e esquerda (76,67%), subclávia esquerda (23,33%), cervical superficial esquerda (3,33%) e do arco aórtico (6,67%).

Irrigação do timo em aves da linhagem Paraíso Pedrês (Gallus gallus domesticus)

A vascularização arterial do timo (número, origem e ordenação) bem como a distribuição parenquimal dos ramos penetrantes foram estudadas em 40 aves da linhagem Paraíso Pedrês. Trinta aves foram injetadas com látex e dissecadas, enquanto 10 aves tiveram seus sistemas arteriais injetados com resina (metil metacrilato e mercox) para a preparação de moldes vasculares. A principal fonte de irrigação encontrada foi a associação dos ramos oriundos das artérias comuns do nervo vago, tireóideas e ingluviais, sendo que seus ramos penetravam o parênquima dos lobos, principalmente pelas suas extremidades cranial e caudal. A partir da penetração, os ramos tímicos apresentavam distribuição predominante para a periferia do lobo, formando uma trama capilar poligonal, com espaços irregulares, característica de um órgão linforreticular.

Molecular basis for porcine parvovirus detection in dead fetuses

Reproductive failures are still common grounds for complaint by commercial swine producers. Porcine parvovirus (PPV) is associated with different clinical reproductive signs. The aim of the present study was to investigate PPV fetal infection at swine farms having ongoing reproductive performance problems. The presence of virus in fetal tissues was determined by nested-polymerase chain reaction assay directed to the conserved NS1 gene of PPV in aborted fetuses, mummies and stillborns. Fetuses show a high frequency of PPV infection (96.4%; N = 28). In 60.7% of the fetuses, PPV were detected in all tissue samples (lung, heart, thymus, kidney, and spleen). Viral infection differed among fetal tissues, with a higher frequency in the lung and heart (P < 0.05). Fetuses with up to 99 days of gestational age and from younger sows showed a higher frequency of PPV (P < 0.05). No significant difference in the presence of PPV was detected among the three clinical presentations. The results suggest that PPV remains an important pathogenic agent associated with porcine fetal death.

Afferent Connections to the Rostrolateral Part of the Periaqueductal Gray: A Critical Region Influencing the Motivation Drive to Hunt and Forage

Previous studies have shown that a particular site in the periaqueductal gray (PAG), the rostrolateral PAG, influences the motivation drive to forage or hunt. To have a deeper understanding on the putative paths involved in the decision-making process between foraging, hunting, and other behavioral responses, in the present investigation, we carried out a systematic analysis of the neural inputs to the rostrolateral PAG (rlPAG), using Fluorogold as a retrograde tracer. According to the present findings, the rlPAG appears to be importantly driven by medial prefrontal cortical areas involved in controlling attention-related and decision-making processes. Moreover, the rlPAG also receives a wealth of information from different amygdalar, hypothalamic, and brainstem sites related to feeding, drinking, or hunting behavioral responses. Therefore, this unique combination of afferent connections puts the rlPAG in a privileged position to influence the motivation drive to choose whether hunting and foraging would be the most appropriate adaptive responses. Copyright (C) 2009 Sandra Regina Mota-Ortiz et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

RNA interference-mediated knockdown of CD49e (alpha 5 integrin chain) in human thymic epithelial cells modulates the expression of multiple genes and decreases thymocyte adhesion

Background: The thymus is a central lymphoid organ, in which bone marrow-derived T cell precursors undergo a complex process of maturation. Developing thymocytes interact with thymic microenvironment in a defined spatial order. A component of thymic microenvironment, the thymic epithelial cells, is crucial for the maturation of T-lymphocytes through cell-cell contact, cell matrix interactions and secretory of cytokines/chemokines. There is evidence that extracellular matrix molecules play a fundamental role in guiding differentiating thymocytes in both cortical and medullary regions of the thymic lobules. The interaction between the integrin alpha 5 beta 1 (CD49e/CD29; VLA-5) and fibronectin is relevant for thymocyte adhesion and migration within the thymic tissue. Our previous results have shown that adhesion of thymocytes to cultured TEC line is enhanced in the presence of fibronectin, and can be blocked with anti-VLA-5 antibody. Results: Herein, we studied the role of CD49e expressed by the human thymic epithelium. For this purpose we knocked down the CD49e by means of RNA interference. This procedure resulted in the modulation of more than 100 genes, some of them coding for other proteins also involved in adhesion of thymocytes; others related to signaling pathways triggered after integrin activation, or even involved in the control of F-actin stress fiber formation. Functionally, we demonstrated that disruption of VLA-5 in human TEC by CD49e-siRNA-induced gene knockdown decreased the ability of TEC to promote thymocyte adhesion. Such a decrease comprised all CD4/CD8-defined thymocyte subsets. Conclusion: Conceptually, our findings unravel the complexity of gene regulation, as regards key genes involved in the heterocellular cell adhesion between developing thymocytes and the major component of the thymic microenvironment, an interaction that is a mandatory event for proper intrathymic T cell differentiation.

Identification of Novel Immunoregulatory Molecules in Human Thymic Regulatory CD4(+)CD25(+) T Cells by Phage Display

Thymic CD4(+)CD25(+) cells play an important role in immune regulation and are continuously developed in the thymus as an independent lineage. How these cells are generated, what are their multiple pathways of suppressive activity and which are their specific markers are questions that remain unanswered. To identify molecules involved in the function and development of human CD4(+)CD25(+) T regulatory cells we targeted thymic CD4(+)CD25(+) cells by peptide phage display. A phage library containing random peptides was screened ex vivo for binding to human thymic CD4(+)CD25(+) T cells. After four rounds of selection on CD4(+)CD25(+) enriched populations of thymocytes, we sequenced several phage displayed peptides and selected one with identity to the Vitamin D Receptor (VDR). We confirmed the binding of the VDR phage to active Vitamin D in vitro, as well as the higher expression of VDR in CD4(+)CD25(+) cells. We suggest that differential expression of VDR on natural Tregs may be related to the relevance of Vitamin D in function and ontogeny of these cells.

DNA oxidation, strand-breaks and etheno-adducts formation promoted by Cu, Zn-superoxide dismutase-H(2)O(2) in the presence and absence of bicarbonate

Biomolecule oxidation promoted by Cu, Zn-superoxide dismutase (SOD1) has been studied because of its potential role in neurodegenerative diseases. We studied the mechanism of DNA damage promoted by the SOD1-H(2)O(2) system. The system promoted the formation of strand breaks in plasmid DNA and the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) in calf thymus DNA. We were also able to detect, for the. first time, 1,N(2)-etheno-2'-deoxyguanosine (1,N(2)-epsilon dGuo) in calf thymus DNA exposed to SOD1-H(2)O(2). The addition of a copper chelator caused a decrease in the frequency of 8-oxodGuo and 1,N(2)-epsilon dGuo, indicating the participation of copper ions lost from SOD1 active sites. The addition of bicarbonate increased the levels of both DNA lesions. We conclude that copper liberated from SOD1 active sites has a central role in the mechanism of DNA damage promoted by SOD1 in the presence of H(2)O(2), and that bicarbonate can modulate the reactivity of released copper.

Contribuition of nitrogen from biological nitrogen fixation, nitrogen fertilizer and soil nitrogen on the growth of the common bean and cowpea

Biological nitrogen fixation (BNF) constitutes a valuable source of this nutrient for the common bean Phaseolus vulgaris L and cowpea Vigna unguiculata (L.) Walp., being its avaibility affected by mineral N in the soil solution. The objectives of this work were to evaluate the effects of nitrogen rate, as urea, on symbiotic fixation of N(2) in common bean and cowpea plants, using the isotopic technique, and quantifying the relative contributions of N sources symbiotic N(2) fixation, soil native nitrogen and urea N on the growth of the common bean and cowpea. Non nodulating soybean plants were used as standard. The research was carried out in greenhouse, using pots with 5 kg of soil from a Typic Haplustox (Dystrophic Red Yellow Latosol). The experimental design was completely randomized blocks, with 30 treatments and three replications, arranged in 5x3x2 factorial outline. The treatments consisted of five N rates: 2, 15, 30, 45 and 60 mg N kg(-1) soil; three sampling times: 23, 40 and 76 days after sowing (DAS) and two crops: common bean and cowpea. The BNF decreased with increase N rates, varying from 81.5% to 55.6% for cowpea, and from 71.9% to 55.1% for common bean. The symbiotic N(2) fixation in cowpea can substitute totally the nitrogen fertilization. The nitrogen absorption from soil is not affected by nitrogen fertilizer rate. The N recovery from fertilizer at 76 DAS was of 60.7% by common bean, and 57.1% by cowpea. The symbiotic association in common bean needs the application of a starting dose (40 kg N ha(-1)) for economically acceptable yields.

Application of molecular techniques to evaluate the methanogenic archaea and anaerobic bacteria in the presence of oxygen with different COD : sulfate ratios in a UASB reactor

In this paper, the microbial characteristics of the granular sludge in the presence of oxygen (3.0 +/- 0.7 mg O-2 1(-1)) were analyzed using molecular biology techniques. The granules were provided by an upflow anaerobic sludge blanket (UASB) operated over 469 days and fed with synthetic substrate. Ethanol and sulfate were added to obtain different COD/SO42- ratios (3.0, 2.0, and 1.6). The results of fluorescent in situ hybridization (FISH) analyses showed that archaeal cells, detected by the ARC915 probe, accounted for 77%, 84%, and 75% in the COD/SO42- ratios (3.0, 2.0, and 1.6, respectively). Methanosaeta sp. was the predominant acetoclastic archaea observed by optical microscopy and FISH analyses, and confirmed by sequencing of the excised bands of the DGGE gel with a similarity of 96%. The sulfate-reducing bacterium Desulfovibrio vulgaris subsp. vulgaris (similarity of 99%) was verified by sequencing of the DGGE band. Others identified microorganism were similar to Shewanella sp. and Desulfitobacterium hafniense, with similarities of 95% and 99%, respectively. These results confirmed that the presence of oxygen did not severely affect the metabolism of microorganisms that are commonly considered strictly anaerobic. We obtained mean efficiencies of organic matter conversion and sulfate reducing higher than 74%. (C) 2008 Elsevier Ltd. All rights reserved.

Photochemical heat-shock response in common bean leaves as affected by previous water deficit

The heat sensitivity of photochemical processes was evaluated in the common bean (Phaseolus vulgaris) cultivars A222, A320, and Carioca grown under well-watered conditions during the entire plant cycle (control treatment) or subjected to a temporal moderate water deficit at the preflowering stage (PWD). The responses of chlorophyll fluorescence to temperature were evaluated in leaf discs excised from control and PWD plants seven days after the complete recovery of plant shoot hydration. Heat treatment was done in the dark (5 min) at the ambient CO2 concentration. Chlorophyll fluorescence was assessed under both dark and light conditions at 25, 35, and 45 degrees C. In the dark, a decline of the potential quantum efficiency of photosystem II (PSII) and an increase in minimum chlorophyll fluorescence were observed in all genotypes at 45 degrees C, but these responses were affected by PWD. In the light, the apparent electron transport rate and the effective quantum efficiency of PSII were reduced by heat stress (45 degrees C), but no change due to PWD was demonstrated. Interestingly, only the A222 cultivar subjected to PWD showed a significant increase in nonphotochemical fluorescence quenching at 45 degrees C. The common bean cultivars had different photochemical sensitivities to heat stress altered by a previous water deficit period. Increased thermal tolerance due to PWD was genotype-dependent and associated with an increase in potential quantum efficiency of PSII at high temperature. Under such conditions, the genotype responsive to PWD treatment enhanced its protective capacity against excessive light energy via increased nonphotochemical quenching.

Macroscopic root water uptake distribution using a matric flux potential approach

Hydrological models featuring root water uptake usually do not include compensation mechanisms such that reductions in uptake from dry layers are compensated by an increase in uptake from wetter layers. We developed a physically based root water uptake model with an implicit compensation mechanism. Based on an expression for the matric flux potential (M) as a function of the distance to the root, and assuming a depth-independent value of M at the root surface, uptake per layer is shown to be a function of layer bulk M, root surface M, and a weighting factor that depends on root length density and root radius. Actual transpiration can be calculated from the sum of layer uptake rates. The proposed reduction function (PRF) was built into the SWAP model, and predictions were compared to those made with the Feddes reduction function (FRF). Simulation results were tested against data from Canada (continuous spring wheat [(Triticum aestivum L.]) and Germany (spring wheat, winter barley [Hordeum vulgare L.], sugarbeet [Beta vulgaris L.], winter wheat rotation). For the Canadian data, the root mean square error of prediction (RMSEP) for water content in the upper soil layers was very similar for FRF and PRF; for the deeper layers, RMSEP was smaller for PRF. For the German data, RMSEP was lower for PRF in the upper layers and was similar for both models in the deeper layers. In conclusion, but dependent on the properties of the data sets available for testing,the incorporation of the new reduction function into SWAP was successful, providing new capabilities for simulating compensated root water uptake without increasing the number of input parameters or degrading model performance.

Extension of the core map of common bean with EST-SSR, RGA, AFLP, and putative functional markers

Microsatellites and gene-derived markers are still underrepresented in the core molecular linkage map of common bean compared to other types of markers. In order to increase the density of the core map, a set of new markers were developed and mapped onto the RIL population derived from the `BAT93` x `Jalo EEP558` cross. The EST-SSR markers were first characterized using a set of 24 bean inbred lines. On average, the polymorphism information content was 0.40 and the mean number of alleles per locus was 2.7. In addition, AFLP and RGA markers based on the NBS-profiling method were developed and a subset of the mapped RGA was sequenced. With the integration of 282 new markers into the common bean core map, we were able to place markers with putative known function in some existing gaps including regions with QTL for resistance to anthracnose and rust. The distribution of the markers over 11 linkage groups is discussed and a newer version of the common bean core linkage map is proposed.

Zn uptake, physiological response and stress attenuation in mycorrhizal jack bean growing in soil with increasing Zn concentrations

The influence of arbuscular mycorrhizal fungi (AMF) inoculation on Canavalia ensiformis growth. nutrient and Zn uptake, and on some physiological parameters in response to increasing soil Zn concentrations was studied. Treatments were applied in seven replicates in a 2 x 4 factorial design, consisting of the inoculation or not with the AMF Glomus etunicatum, and the addition of Zn to soil at the concentrations of 0, 100, 300 and 900 mg kg(-1). AMF inoculation enhanced the accumulation of Zn in tissues and promoted biomass yields and root nodulation. Mycorrhizal plants exhibited relative tolerance to Zn up to 300 mg kg(-1) without exhibiting visual symptoms of toxicity, in contrast to non-mycorrhizal plants which exhibited a significant growth reduction at the same soil Zn concentration. The highest concentration of Zn added to soil was highly toxic to the plants. Leaves of plants grown in high Zn concentration exhibited a Zn-induced proline accumulation and also an increase in soluble amino acid contents; however proline contents were lower in mycorrhizal jack beans. Plants in association or not with the AMF exhibited marked differences in the foliar soluble amino acid profile and composition in response to Zn addition to soil. In general, Zn induced oxidative stress which could be verified by increased lipid peroxidation rates and changes in catalase, ascorbate peroxidase, glutathione reductase and superoxide dismutase activities. In summary, G. etunicatum was able to maintain an efficient symbiosis with jack bean plants in moderately contaminated Zn-soils, improving plant performance under those conditions, which is likely to be due to a combination of physiological and nutritional changes caused by the intimate relation between fungus and plant. The enhanced Zn uptake by AMF inoculated jack bean plants might be of interest for phytoremediation purposes. (C) 2009 Elsevier Ltd. All rights reserved.

Acquired tolerance of tomato (Lycopersicon esculentum cv. Micro-Tom) plants to cadmium-induced stress

The effects of varying concentrations of cadmium (Cd) on the development of Lycopersicon esculentum cv. Micro-Tom (MT) plants were investigated after 40 days (vegetative growth) and 95 days (fruit production), corresponding to 20 days and 75 days of exposure to CdCl(2), respectively. Inhibition of growth was clearly observed in the leaves after 20 days and was greater after 75 days of growth in 1 mM CdCl(2), whereas the fruits exhibited reduced growth following the exposure to a concentration as low as 0.1 mM CdCl(2). Cd was shown to accumulate in the roots after 75 days of growth but was mainly translocated to the upper parts of the plants accumulating to high concentrations in the fruits. Lipid peroxidation was more pronounced in the roots even at 0.05 mM CdCl(2) after 75 days, whereas in leaves, there was a major increase after 20 days of exposure to 1 mM CdCl(2), but the fruit only exhibited a slight significant increase in lipid peroxidation in plants subjected to 1 mM CdCl(2) when compared with the control. Oxidative stress was also investigated by the analysis of four key antioxidant enzymes, which exhibited changes in response to the increasing concentrations of Cd tested. Catalase (EC 1.11.1.6) activity was shown to increase after 75 days of Cd treatment, but the major increases were observed at 0.1 and 0.2 mM CdCl(2), whereas guaiacol peroxidase (EC 1.11.1.7) did not vary significantly from the control in leaves and roots apart from specific changes at 0.5 and 1 mM CdCl(2). The other two enzymes tested, glutathione reductase (EC 1.6.4.2) and superoxide dismutase (SOD, EC 1.15.1.1), did not exhibit any significant changes in activity, apart from a slight decrease in SOD activity at concentrations above 0.2 mM CdCl(2). However, the most striking results were obtained when an extra treatment was used in which a set of plants was subjected to a stepwise increase in CdCl(2) from 0.05 to 1 mM, leading to tolerance of the Cd applied even at the final highest concentration of 1 mM. This apparent adaptation to the toxic effect of Cd was confirmed by biomass values being similar to the control, indicating a tolerance to Cd acquired by the MT plants.

Validation of a root water uptake model to estimate transpiration constraints

Experimental results obtained from a greenhouse trial with common bean (Phaseolus vulgaris L) plants performed to test model hypotheses regarding the onset of limiting hydraulic conditions and the shape of the transpiration reduction curve in the falling rate phase are presented. According to these hypotheses based on simulations with an upscaled single-root model, the matric flux potential at the onset of limiting hydraulic conditions is as a function of root length density and potential transpiration rate, while the relative transpiration in the falling rate phase equals the relative matric flux potential. Transpiration of bean plants in water stressed pots with four different soils was determined daily by weighing and compared to values obtained from non-stressed pots. This procedure allowed determining the onset of the falling rate phase and corresponding soil hydraulic conditions. At the onset of the falling rate phase, the value of matric flux potential M(I) showed to differ in order of magnitude from the model predicted value for three out of four soils. This difference between model and experiment can be explained by the heterogeneity of the root distribution which is not considered by the model. An empirical factor to deal with this heterogeneity should be included in the model to improve predictions. Comparing the predictions of relative transpiration in the falling rate phase using a linear shape with water content, pressure head or matric flux potential, the matric flux potential based reduction function, in agreement with the hypothesis, showed the best performance, while the pressure head based equation resulted in the highest deviations between observed and predicted values of relative transpiration rates. (C) 2010 Elsevier B.V. All rights reserved.