Pivotal Role for Platelet-Activating Factor Receptor in CD36 Expression and oxLDL Uptake by Human Monocytes/Macrophages

The uptake of oxLDL by CD36 is not regulated by intracellular levels of cholesterol, leading to macrophage differentiation into foam cells which play a major role in atherosclerosis. Furthermore, oxLDL competes with PAF in macrophages for binding to PAF receptors (PAFR). Here we investigated the involvement of PAFR in CD36 expression and uptake of oxLDL by human monocytes/macrophages. Adherent peripheral blood mononuclear cells were treated with PAFR-antagonists (WEB2170, CV3988); inhibitors of ERK1/2 (PD98059), p38 (SB203580), JNK (SP600125) or diluents, before stimulation with oxLDL or PAF. After 24 h, uptake of FITC oxLDL and expression of CD36 was determined by flow cytometry and phosphorylation of MAP-kinases by Western blot. It was shown that the uptake of oxLDL was reduced by PAFR antagonists. CD36 expression was up-regulated by oxLDL, an effect reversed by PAFR antagonists. The up-regulation of CD36 and oxLDL uptake both required MAP-kinases activation. The oxLDL induced ERK1/2 and JNK but not p38 phosphorylation was reversed by PAFR-antagonists suggesting that oxLDL signalling involves PAFR dependent and independent pathways. In macrophages from PAFR(-/-) mice, oxLDL was unable to up-regulate CD36 expression and the oxLDL uptake was reduced compared to wild type. These results suggest that oxLDL interacts with PAFR in macrophages to increase CD36 expression and oxLDL uptake. Whereas pharmacological intervention at the level of PAFR would be beneficial in atherosclerosis remains to be determined. Copyright (C) 2011 S. Karger AG, Basel

L-Proline uptake in Crithidia deanei is influenced by its endosymbiont bacterium

Crithidia deanei, a monoxenic trypanosomatid, presents an endosymbiotic bacterium in its cytoplasm. Both the protozoan and the bacterium maintain intensive metabolic exchange, resulting in an interesting model to study the coevolution of metabolisms. The relevance of L-proline for the growth of C. deanei and its transport into these cells was studied. Both the endosymbiont-containing (wild) and the endosymbiont-free protozoa (aposymbiont or cured) strains, when grown in medium supplemented with L-proline, reached higher cell densities than those grown in unsupplemented media. We biochemically characterized the uptake of L-proline in both the wild (K(m)=0.153 +/- 0.022 mM, V(max)=0.239 +/- 0.011 nmol min(-1) per 4 x 10(7) cells) and the aposymbiont strains (K(m)=0.177 +/- 0.049 mM, V(max)=0.132 +/- 0.012 nmol min(-1) per 4 x 10(7) cells). These data suggest a single type of proline transporter whose activity is upregulated by the presence of the symbiotic bacterium. Proline transport was further characterized and was found to be insensitive to the extracellular concentration of Na(+), but sensitive to K(+) and pH. The abolition of proline uptake by respiratory chain inhibitors and valinomycin indicates that the proline transport in C. deanei is dependent on the plasma membrane K(+) gradient.

Root uptake and reduction of hexavalent chromium by aquatic macrophytes as assessed by high-resolution X-ray emission

Aquatic macrophytes Salvinia auriculata, Pistia stratiotes and Eichhornia crassipes were chosen to investigate the Cr(VI) reduced by root-based biosorption in a chromium uptake experiment, using a high-resolution XRF technique. These plants were grown in hydroponics medium supplied with non-toxic Cr concentrations during a 27-day metal uptake experiment. The high-resolution Cr-K beta fluorescence spectra for dried root tissues and Cr reference material (100% Cr, Cr(2)O(3), and CrO(3)) were measured using an XRF spectrometer. For all species of aquatic plant treated with Cr(VI), the energy of the Cr-K beta(2,5) line was shifted around 8 eV below the same spectral line identified for the Cr(VI) reference, but it was also near to the line identified for the Cr(III) reference. Moreover, there was a lack of the strong Cr-K beta"" line assigned to the Cr(VI) reference material within the Cr(VI)-treated plant spectra, suggesting the reduction of Cr(VI) for other less toxic oxidation states of Cr. As all Cr-K beta spectra of root tissue species were compared, the peak energies and lineshape patterns of the Cr-K beta(2,5) line are coincident for the same aquatic plant species, when they were treated with Cr(III) and Cr(VI). Based on the experimental evidence, the Cr(VI) reduction process has happened during metal biosorption by these plants. (C) 2009 Elsevier Ltd. All rights reserved.

Glucose uptake in the mammalian stages of Trypanosoma cruzi

Trypanosoma cruzi, the agent of Chagas` disease, alternates between different morphogenetic stages that face distinct physiological conditions in their invertebrate and vertebrate hosts, likely in the availability of glucose. While the glucose transport is well characterized in epimastigotes of T cruzi, nothing is known about how the mammalian stages acquire this molecule. Herein glucose transport activity and expression were analyzed in the three developmental stages present in the vertebrate cycle of T cruzi. The infective trypomastigotes showed the highest transport activity (V(max) = 5.34 +/- 0.54 nmol/min per mg of protein: K(m) = 0.38 +/- 0.01 mM) when compared to intracellular epimastigotes (V(max) = 2.18 +/- 0.20 nmol/min per mg of protein; K(m) = 0.39 +/- 0.01 mM). Under the conditions employed no transport activity could be detected in amastigotes. The gene of the glucose transporter is expressed at the mRNA level in trypomastigotes and in intracellular epimastigotes but not in amastigotes, as revealed by real-time PCR. In both trypomastigotes and intracellular epimastigotes protein expression could be detected by Western blot with an antibody raised against the glucose transporter correlating well with the transport activity measured experimentally. Interestingly, anti-glucose transporter antibodies showed a strong reactivity with glycosome and reservosome organelles. A comparison between proline and glucose transport among the intracellular differentiation forms is presented. The data suggest that the regulation of glucose transporter reflects different energy and carbon requirements along the intracellular life cycle of T cruzi. (C) 2009 Elsevier B.V. All rights reserved.

The GATA-type transcriptional activator Gat1 regulates nitrogen uptake and metabolism in the human pathogen Cryptococcus neoformans

Nitrogen uptake and metabolism are essential to microbial growth. Gat1 belongs to a conserved family of zinc finger containing transcriptional regulators known as GATA-factors. These factors activate the transcription of Nitrogen Catabolite Repression (NCR) sensitive genes when preferred nitrogen sources are absent or limiting. Cryptococcus neoformans GAT1 is an ortholog to the Aspergillus nidulans AreA and Candida albicans GAD genes. In an attempt to define the function of this transcriptional regulator in C. neoformans, we generated null mutants (gat1 Delta) of this gene. The gat 1 mutant exhibited impaired growth on all amino acids tested as sole nitrogen sources, with the exception of arginine and proline. Furthermore, the gat1 mutant did not display resistance to rapamycin, an immunosuppressant drug that transiently mimics a low-quality nitrogen source. Gal is not required for C. neoformans survival during macrophage infection or for virulence in a mouse model of cryptococcosis. Microarray analysis allowed the identification of target genes that are regulated by Gat1 in the presence of proline, a poor and non-repressing nitrogen source. Genes involved in ergosterol biosynthesis, iron uptake, cell wall organization and capsule biosynthesis, in addition to NCR-sensitive genes, are Gat1-regulated in C. neoformans. (C) 2010 Elsevier Inc. All rights reserved.

Evaluation of some metals in Brazilian coffees cultivated during the process of conversion from conventional to organic agriculture

The aim of this study was to evaluate the presence of nutrients and toxic elements in coffees cultivated during the process of conversion, on organic agriculture, in southwest Bahia, Brazil. Levels of the nutrients and toxic elements were determined in samples of soils and coffee tissues from two transitional organic farms by atomic absorption spectrometry (FAAS). The metals in soil samples were extracted by Mehlich1 and USEPA-3050 procedures. Coffee samples from both farms presented relatively high levels of Cd, Zn and Cu (0.75,45.4 and 14.9 mu g g(-1). respectively), but were still below the limits specified by the Brazilian Food Legislation. The application of statistical methods showed that this finding can be attributed to the addition of high amounts of organic matter during the flowering tree period which can act on the bioavailability of metal ions in soils. (C) 2009 Elsevier Ltd. All rights reserved.

Estudo do crescimento, eficiência de biofiltração e cinética de absorção de nutrientes (N-NH, N-NO e P-PO4³) da macroalga Gracilaria cervicornis (Turner) J. Agardh

The objective of this study was to examine the growth of Gracilaria cervicornis cultured in a shrimp (Litopenaeus vannamei) pond and to determine the absorption efficiency and the kinetics parameters (Vmax, Ks e Vmax:Ks) of this macroalgae for the nutrients N-NO3-, N-NH4+ and P-PO4-3, aiming at its use as bioremediatory of eutrophicated environments. For this study, two experiments (field and laboratory) were developed. In the field study, the seaweed was examined in relation to the growth and the biomass. In the laboratory experiment, the absorption efficiency of G. cervicornis was measured through the monitoring of the concentration of the three nutrients (N-NO3-, N-NH4+ e P-PO4-3) during 5 hours and the kinetic parameters were determined through the formula of Michaelis-Menten. The results obtained in this study demonstrated that G. cervicornis benefited from the available nutrients in the pond, increasing 52.4% of its biomass value after 30 days of culture. It was evidenced that the variability of the biomass could be explained through the salinity, availability of light (transparency and solid particle in suspension) and concentration of N-NO3- in the environment. In the laboratory experiment, the highest absorption efficiency was found in the treatments with low concentration (5 µmol.L-1), being evidenced a reduction of up to 85,3%, 97,5% and 81,2% of N-NH4+, N-NO3- and P-PO43-, respectively. Regarding the kinetic parameters, G. cervicornis presented better ability in absorbing N-NH4+ in high concentrations (Vmax = 158,5 µmol g-1 dry wt h-1) and P-PO43- in low concentrations (Ks = 5 µmol.L-1 e Vmax:Ks = 10,3). The results of this study show that G. cervicornis could be cultivated in shrimp ponds, presents a good capacity of absorption for the tested nutrients and is a promising candidate for biorremediation in shrimp pond effluent

Peat humic substances enriched with nutrients for agricultural applications: Competition between nutrients and non-essential metals present in tropical soils

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Selenite and Selenate Effects on Mercury (Hg2+) Uptake and Distribution in Tilapia, Oreochromis niloticus L., Assessed by Chronic Bioassay

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

Accumulation of nutrients in sweet peppers cultivated in coconut fiber

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

Crescimento e acúmulo de nutrientes em couve-flor Verona

This work consisted in an experiment installed at Unesp, Campus of Jaboticabal, from January to May 2006 and the objective was to evaluate dry matter accumulated and nutrients by cauliflower, hybrid Verona, during the cycle. The experimental design was randomized blocks, in plots subdivided on the time, with three replications. The plants were sampled at 30, 45, 60, 75, 90, 105 and 120 days after sowing (DAS), and were evaluated: number of leaves, stem length, stem and curd diameter, leaf area, stem, leaves and curd dry matter and accumulate of nutrients. The cauliflower growth had four different phases. In the first one, from 0 to 30 DAS, was correspondent of the period from sowing to transplant. In the second phase, from 30 to 75 DAS, the dry matter accumulated in leaves and stem were small in comparison with the total. The third growth phase, from 75 to 97 DAS, was characterized with great dry matter accumulated, increase in the number of leaves and leaf area. This phase ended with visualization of the curd, and was the beginning to the fourth phase, from 97 to 120 DAS. In the culture estimated in 25,000 plants/ha and productivity of 25.2 t ha(-1) the accumulated quantities with of N, P, K, Ca, Mg and S were, corresponding to, 224.5; 54; 156; 136; 21.75; 63.5 kg ha(-1) and 714.8; 270; 236.7 and 33.3 g ha(-1), corresponding to Fe, Mn, Zn and Cu.

Curva de crescimento e marcha de absorção de macronutrientes em três cultivares de amoreira (Morus alba L)

To evaluate the growth and pattern of nutrient absorption (N, P, K, Ca, Mg and S amongst others) in three berry cultivars (treatments), four plants by treatment in different phonological stages were harvested. The experiment was arranged in randomized design of factorial 3x5 with three replicates. There cultivars were used FM Shima-Miura, IZ 56/4 and Miura and above ground biomass was harvested in five occasions 45, 60, 75, 90 and 115 days after plantation. It was observed that the accumulation of nutrients presented different pattern and cv. IZ 56/4 was superior compared with PM Shima-Miura for N, P, K, Ca, Mg and S and N, P, K and S for Miura. For example, the accumulation of N expressed on mg plant"(1) per berry plant cultivar PM Shima-miura, IZ 56/4 and Miura was respectively 4,21; 8,14 and 5,56, for K 2,71; 5,22 and 2,64 and for Ca 2,47; 4,52 and 3,59.

Produção de matéria seca e absorção de nutrientes pelo milho em razão da saturação por bases e da adubação potássica

O trabalho foi conduzido em condições de casa de vegetação, com o objetivo de avaliar a resposta do milho à adubação potássica e à saturação do solo por bases, na produção de matéria seca e na absorção de nutrientes. O delineamento experimental foi inteiramente casualizado, com quatro repetições, em esquema fatorial 3x2x4, constituindo-se de três solos (Areia Quartzosa, Latossolo Vermelho-Escuro e Latossolo Roxo), dois valores de saturação por bases (40% e 70 %) e quatro teores de K no solo (0,5, 1,5, 3,0 e 6,0 mmol c dm-3). Para a calagem, foi utilizada mistura de carbonato de cálcio + carbonato de magnésio (PRNT = 103,3 %) na proporção de 4:1. A adubação básica constou de 200 mg kg-1 de N, 200 mg kg-1 de P e 5 mg kg-1 de Zn por vaso de 30 L, sendo o N parcelado em 2,48 g (83,7 mg kg-1) na semeadura e o restante em duas coberturas aos 25 e 40 dias após a emergência das plântulas (DAE). As doses de K utilizadas foram de 0, 3,62, 7,24 e 14,48 g de KCl por vaso. A semeadura foi realizada em 4/3/97, utilizando-se o milho cv. Zêneca 8392, mantendo-se uma planta/vaso durante 60 DAE. O K proporciona ganhos de matéria seca até teores de 1,5 mmol c dm-3 no solo. O aumento dos teores de K no solo resultam em queda na concentração de Ca e Mg na matéria seca do milho. A elevação da saturação por bases e respectivo aumento dos teores de Ca e Mg no solo reduzem a absorção de K pelo milho.

Produção de mudas de caramboleiras 'B-10' e 'Golden Star': I - parâmetros biológicos

O Brasil é um dos maiores produtores de carambola do mundo, entretanto há poucas informações científicas, especialmente estudos de nutrição mineral com mudas dessa frutífera. Objetivando contribuir com o conhecimento desse importante aspecto, desenvolveu-se estudo que permitisse avaliar o crescimento e o acúmulo de nutrientes em mudas de caramboleiras, cultivadas em solução nutritiva. O experimento foi realizado em parcelas subdivididas, sendo utilizadas como parcela as duas cultivares de caramboleira ('B-10' e 'Golden Star') e, como subparcelas, as cinco épocas de coleta de plantas, realizadas aos 208; 233; 258; 283 e 308 dias após o transplantio para a solução nutritiva. O delineamento foi inteiramente casualizado, com três repetições. As mudas foram cultivadas em vasos (8L) com solução nutritiva (pH=5,5 ± 0,5), com aeração. O experimento iniciou-se em 24-08-2005. Nos diferentes órgãos das mudas (folhas, caule e raízes), avaliaram-se o crescimento e o acúmulo de nutrientes, e os índices nutricionais. Não houve diferenças no crescimento e, em geral, no acúmulo da massa da matéria seca entre as duas cultivares. Houve acúmulo linear da massa da matéria seca das mudas de caramboleira com o tempo de cultivo, sendo maior nas folhas > caule > raízes. O período de maior acúmulo da massa de matéria seca e da taxa de crescimento relativo na planta inteira esteve compreendido entre 208 - 233 e 233 - 258 dias após o transplantio para a 'B-10' e a 'Golden Star', respectivamente.

Acúmulo de massa seca e marcha de absorção de nutrientes em mudas de goiabeira 'Pedro Sato'

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