The polysaccharide fraction of Propionibacterium acnes modulates the development of experimental focal segmental glomerulosclerosis

The pathogenesis of focal segmental glomerulosclerosis (FSGS) appears to be associated with type-2 cytokines and podocyte dysfunction. In this study, we tested the hypothesis that immunization with the polysaccharide fraction of Propionibacterium acnes (PS), a pro-Th1 agonist, may subvert the type-2 profile and protect podocytes from adriamycin-induced glomerulosclerosis. Adriamycin injection resulted in albuminuria and increased serum creatinine in association with loss of glomerular podocin and podoplanin expression, which is consistent with podocyte dysfunction. Renal tissue analysis revealed the expression of transcripts for GATA3 and fibrogenic-related proteins, such as TGF-beta, tissue inhibitor of metalloproteinase-1 (TIMP-1) and metalloproteinase 9 (MMP9). In association with the expression of fibrogenic transcripts, we observed peri-glomerular expression of a-smooth muscle actin (alpha-SMA), indicating epithelial-to-mesenchymal transition, and increased expression of proliferating cell nuclear antigen (PCNA) in tubular cells, suggesting intense proliferative activity. Previous immunization with PS inhibited albuminuria and serum creatinine in association with the preservation of podocyte proteins and inhibition of fibrogenic transcripts and the expression of alpha-SMA and PCNA proteins. Tissue analysis also revealed that PS treatment induced expression of mRNA for GD3 synthase, which is a glycosiltransferase related to the synthesis of GD3, a ganglioside associated with podocyte physiology. In addition, PS treatment inhibited the influx of inflammatory CD8(pos) and CD11b(pos) cells to kidney tissue. Finally, PS treatment on day 4 post-ADM, a period when proteinuria was already established, was able to improve renal function. Thus, we demonstrate that the PS fraction of P. acnes can inhibit FSGS pathogenesis, suggesting that immunomodulation can represent an alternative approach for disease management. (C) 2011 Elsevier GmbH. All rights reserved.

Physiological effects of glyphosate over amino acid profile in conventional and transgenic soybean (Glycine max)

This paper compares the responses of conventional and transgenic soybean to glyphosate application in terms of the contents of 17 detectable soluble amino acids in leaves, analyzed by HPLC and fluorescence detection. Glutamate, histidine, asparagine, arginine + alanine, glycine + threonine and isoleucine increased in conventional soybean leaves when compared to transgenic soybean leaves, whereas for other amino acids, no significant differences were recorded. Univariate analysis allowed us to make an approximate differentiation between conventional and transgenic lines, observing the changes of some variables by glyphosate application. In addition, by means of the multivariate analysis, using principal components analysis (PCA), cluster analysis (CA) and linear discriminant analysis (LDA) it was possible to identify and discriminate different groups based on the soybean genetic origin. (C) 2011 Elsevier Inc. All rights reserved.

Genotoxicity assessment of water soluble fractions of biodiesel and its diesel blends using the Salmonella assay and the in vitro MicroFlow (R) kit (Litron) assay

The designation of biodiesel as an environmental-friendly alternative to diesel oil has improved its commercialization and use. However, most biodiesel environmental safety studies refer to air pollution and so far there have been very few literature data about its impacts upon other biotic systems, e.g. water, and exposed organisms. Spill simulations in water were carried out with neat diesel and biodiesel and their blends aiming at assessing their genotoxic potentials should there be contaminations of water systems. The water soluble fractions (WSF) from the spill simulations were submitted to solid phase extraction with C-18 cartridge and the extracts obtained were evaluated carrying out genotoxic and mutagenic bioassays [the Salmonella assay and the in vitro MicroFlow (R) kit (Litron) assay]. Mutagenic and genotoxic effects were observed, respectively, in the Salmonella/microsome preincubation assay and the in vitro MN test carried out with the biodiesel WSF. This interesting result may be related to the presence of pollutants in biodiesel derived from the raw material source used in its production chain. The data showed that care while using biodiesel should be taken to avoid harmful effects on living organisms in cases of water pollution. (C) 2011 Elsevier Ltd. All rights reserved.

Interaction of cationic water-soluble meso-tetrakis(4-N-methylpyridiniumyl)porphyrin (TMPyP) with ionic and nonionic micelles: aggregation and binding

The equilibrium of meso-tetrakis(4-N-methylpyridiniumyl)porphyrin (TMPyP) in aqueous solution in the presence of surfactants was studied by optical spectroscopic techniques and SAXS (small angle X-ray scattering). Anionic SDS (sodium dodecyl sulfate), zwitterionic HPS (N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate) and nonionic TRITON X-100 (t-octyl-phenoxypolyethoxyethanol), surfactants were used. TMPyP is characterized by a protonation equilibrium with a pK(a) around 1.0, associated with the diacid-free base transition, and a second pK(a) around 12.0 related with the transition between the free base and the monoanion form. Three independent species were observed for TMPyP at pH 6.0 as a function of SDS concentration: free TMPyP, TMPyP-SDS aggregates and porphyrin monomer bound to micelles. For HPS and TRITON X-100, the equilibrium of TMPyP as a function of pH is quite similar to that obtained in pure aqueous solution: no aggregation was observed, suggesting that electrostatic contribution is the major factor in the interaction between TMPyP and surfactants. SAXS data analysis demonstrated a prolate ellipsoidal shape for SDS micelles; no significant changes in shape and size were observed for SDS-TMPyP co-micelles. Moreover, the ionization coefficient, alpha, decreases with the increase of the porphyrin concentration, suggesting the ""screening"" of the anionic charge of SDS by the cationic porphyrin. These results are consistent with optical absorption, fluorescence and RLS (resonance light scattering) spectroscopies data, allowing to conclude that neutral surfactants present a smaller interaction with the cationic porphyrin as compared with an ionic surfactant. Therefore, the interaction of TMPyP with the ionic and nonionic surfactants is predominantly due to the electrostatic contribution. Copyright (c) 2008 Society of Porphyrins & Phthalocyanines.

Expression of soluble and functional full-length human matrix metalloproteinase-2 in Escherichia coli

Characterization of the matrix metalloproteinase-2 (MMP-2) substrates and understanding of its function remain difficult because up to date preparations containing minor amounts of other eukaryotic proteins that are co-purified with MMP-2 are still used. In this work, the expression of a soluble and functional full-length recombinant human MMP-2 (rhMMP-2) in the cytoplasm of Escherichia coli is reported, and the purification of this metalloproteinase is described. Culture of this bacterium at 18 degrees C culminated in maintenance of the soluble and functional rhMMP-2 in the soluble fraction of the E. coli lysate and its purification by affinity with gelatin-sepharose yielded approximately 0.12 mg/L of medium. Western Blotting and zymographic analysis revealed that the most abundant form was the 72-kDa MMP-2, but some gelatinolytic bands corresponding to proteins with lower molecular weight were also detected. The obtained rhMMP-2 was demonstrated to be functional in a gelatinolytic fluorimetric assay, suggesting that the purified rhMMP-2 was correctly folded. The method described here involves fewer steps, is less expensive, and is less prone to contamination with other proteinases and MMP inhibitors as compared to expression of rhMMP-2 in eukaryotic tissue culture. This protocol will facilitate the use of the full-length rhMMP-2 expressed in bacteria and will certainly help researchers to acquire new knowledge about the substrates and biological activities of this important proteinase. (C) 2011 Elsevier B.V. All rights reserved.

Trichogramma pretiosum parasitism and dispersal capacity: a basis for developing biological control programs for soybean caterpillars

In order to succeed in biological control programs, not only is it crucial to understand the number of natural enemies to be released but also on how many sites per area this releasing must be performed. These variables might differ deeply among egg parasitoid species and crops worked. Therefore, these trials were carried out to evaluate the parasitism (%) in eggs of Anticarsia gemmatalis and Pseudoplusia includens after the release of different densities of the egg parasitoid Trichogramma pretiosum. Field dispersal was also studied, in order to determine appropriate recommendations for the release of this parasitoid in soybean fields. The regression analysis between parasitism (%) and densities of the parasitoid indicated a quadratic effect for both A. gemmatalis and P. includens. The maximum parasitism within 24 h after the release was reached with densities of 25.6 and 51.2 parasitoids per host egg, respectively, for the two pests. Parasitism of T. pretiosum in eggs of P. includens decreased linearly as the distance of the pest eggs from the parasitoid release sites increased. For P. includens, the mean radius of T. pretiosum action and the area of parasitoid dispersal in the soybean crop were 8.01 m and 85.18 m(2), respectively. We conclude that for a successful biological control program of lepidopteran pests using T. pretiosum in soybean fields, a density of 25.6 parasitoids per host egg, divided into 117 sites per hectare, should be used.

Rice oil as a soybean oil replacement in weanling pig diets

The motivation for including oils in the diets of weanling pigs is to increase dietary energy density to meet the pigs' energy requirements. The purpose of this study was to determine the digestible and metabolizable energy values of semi-refined rice oil (RO) and refined soybean oil (SO) for weanling pigs (Experiment 1), as well as to evaluate how replacing SO with RO in weanling pig diets affects performance, intestinal histology, organ morphometry and the occurrence of diarrhea (Experiment 2). In Experiment 1,21 barrows, averaging 19.63 +/- 0.37 kg body weight (BW), were used in a randomized complete block design experiment, with seven replications per treatment and one pig per experimental unit (metabolism cage). The treatments consisted of a reference diet and two test diets. The test diets were obtained by using 90% of the reference diet and 10% of RO or SO. The experimental period consisted of a 7 days adaptation period, followed by a 5 days collection period of feces and urine. The values of the apparent digestible energy, apparent metabolizable energy and N-corrected apparent metabolizable energy of the oils were, respectively, 26.24, 25.78 and 26.01 MJ kg(-1) for RO and 31.74, 31.35 and 31.51 MJ kg(-1) for SO. In Experiment 2, 120 weaned pigs, castrated males and females, averaging 6.74 +/- 0.42 kg BW, were used in a randomized complete block design experiment, with eight replications per treatment and three pigs per experimental unit (pen) to evaluate the effects of five replacement levels of SO with RO in isonutritive weanling pig diets. The treatments were as follows: 4:0 basal diet with 4% of SO; 3:1 basal diet with 3% of SO and 1% of RO; 2:2 basal diet with 2% of SO and 2% of RO; 1:3 basal diet with 1% of SO and 3% of RO; and 0:4 basal diet with 4% of RO. No effects of the replacement levels were observed in the weanling pigs' performance, organ morphometry or in their occurrence of diarrhea. However, a quadratic effect on the villus width and a cubic effect on the villus height:crypt depth ratio were observed in the duodenum. Therefore, if the determined values of energy are used in the feed formulation, refined soybean oil can be replaced with semi-refined rice oil in weanling pig diets without affecting their growth performance. (C) 2011 Elsevier B.V. All rights reserved.

Assessment of the high-dose concept and level of control provided by MON 87701 x MON 89788 soybean against Anticarsia gemmatalis and Pseudoplusia includens (Lepidoptera: Noctuidae) in Brazil

BACKGROUND: Genetically modified MON 87701 X MON 89788 soybean (Glycine max), which expresses the Cry1Ac and EPSP-synthase proteins, has been registered for commercial use in Brazil. To develop an Insect Resistance Management (IRM) program for this event, laboratory and field studies were conducted to assess the high-dose concept and level of control it provides against Anticarsia gemmatalis and Pseudoplusia includens. RESULTS: The purified Cry1Ac protein was more active against A. gemmatalis [LC50 (FL 95%) = 0.23 (0.150.34) mu g Cry1Ac mL-1 diet] than P. includens [LC50 (FL 95%) = 3.72 (2.654.86) mu g Cry1Ac mL-1 diet]. In bioassays with freeze-dried MON 87701X MON 89788 soybean tissue diluted 25 times in an artificial diet, there was 100% mortality of A. gemmatalis and up to 95.79% mortality for P. includens. In leaf-disc bioassays and under conditions of high artificial infestation in the greenhouse and natural infestation in the field, MON 87701X MON 89788 soybean showed a high level of efficacy against both target pests. CONCLUSIONS: The MON 87701X MON 89788 soybean provides a high level of control against A. gemmatalis and P. includes, but a high-dose event only to A. gemmatalis. Copyright (c) 2012 Society of Chemical Industry

Thyroid hormones are involved in 5 '-nucleotidase modulation in soluble fraction of cardiac tissue

Aims: To investigate the role of TH (thyroid hormones) in 5'-nucleotidase activity and expression in cardiac soluble fraction (SF). Main methods: Male Wistar rats received daily injections of 14 (10, 25 or 50 mu g T4/100 g body weight) for 14 days to develop a hyperthyroidism condition. Thyroidectomy was performed in other animals to mimic hypothyroidism, and 14 days after surgery they were submitted to TH replacement therapy. Key findings: T4 reduced the 5'-nucleotidase activity (T4-25. P<0.05 and 14-50, P<0.01) in the SF. Conversely, hypothyroidism significantly increased the 5'-nucleotidase activity in this fraction (P<0.001) and TH replacement therapy reversed the latter result (P<0.001 compared to hypothyroid group). The analysis of protein expression in the SF showed that 5'-nucleotidase was more expressed in hypothyroid than in the control group and that the phosphorylated state of PKC observed in this condition may contribute to a possible mechanism of 5'-nucleotidase modulation by thyroid status. Significance: Taken together, these data reveal that TH can influence adenosine production by modulating 5'-nucleotidase activity and expression, which may contribute to the cardioprotective effect and the maintenance of cardiac function under TH privation. (C) 2012 Elsevier Inc. All rights reserved.

Exploring the Role of Soluble Factors Associated with Immune Regulatory Properties of Mesenchymal Stem Cells

Mesenchymal stem cells (MSCs) are characterized as multipotent stromal cells with the capacity for both self-renewal and differentiation into mesodermal cell lineages. MSCs also have a fibroblast-like phenotype and can be isolated from several tissues. In recent years, researchers have found that MSCs secrete several soluble factors that exert immunosuppressive effects by modulating both innate (macrophages, dendritic and NK cells) and adaptive (B cells and CD4+ and CD8+ T cells) immune responses. This review summarizes the principal trophic factors that are related to immune regulation and secreted by MSCs under both autoimmune and inflammatory conditions. The understanding of mechanisms that regulate immunity in MSCs field is important for their future use as a novel cellular-based immunotherapy with clinical applications in several diseases.

Reciprocal gene flow between conventional and genetically modified soybean cultivars

The objective of this work was to determine the reciprocal gene flow between two soybean cultivars, one tolerant and the other sensitive to glyphosate, as well as to use estimators to determine the outcrossing rate in the population and the number of hybrid seeds in the progeny. The experiment was composed of four blocks of 40 soybean rows, of which 20 rows of each cultivar (CD217 and CD219RR). At the R8 stage, five rows, distant 0.5, 1, 2, 4 and 8 m from the adjacent cultivar, were harvested, threshed and analyzed as for the occurrence of gene flow. As phenotypical markers, the trait color of flowers, hypocotil and pubescence, as well as the tolerance to glyphosate were used. The cultivars contrast for all the analyzed traits, each one conditioned by a single gene with two alleles, in a complete dominance interaction. In the tolerant cultivar progeny, the largest outcross rate was 0.27%, and in the sensitive cultivar progeny, 0.83% was identified; by the dilution effect hypothesis, the outcross rates in natural populations would be 0.104 and 0.388%, respectively. The reciprocal gene flow between CD217 and CD219RR cultivars is not the same in both directions. The proposed estimators are useful for determining the hybrid rates in seed samples.

Drospirenone and levonorgestrel in combination with either 30 or 20 mcg ethinylestradiol reduce soluble adhesion molecules in Brazilian women; cross-sectional study

Background: The objective of this study was to evaluate the effect of three contraceptive pills containing ethinylestradiol (EE) (20 or 30 mcg) in combination with drospirenone (DRSP) and levonorgestrel (LNG) on plasma concentration of adhesion molecules vascular cell adhesion molecule -1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) and E-selectin. Study Design: A cross-sectional study was conducted with 72 participants (18-30 years old) distributed into three groups that used oral contraceptives containing EE 20 or 30 mcg combined with DRSP 3 mg or EE 30 mcg/LNG 150 mcg for at least 6 months. The control group was comprised of nonusers of contraceptives. Soluble VCAM-1, soluble ICAM-1 and soluble E-selectin were evaluated by enzyme-linked immunosorbent assay. Results: Compared to the control group, a significant decrease was found in VCAM-1 and ICAM-1 concentrations with use of DRSP/20 EE and LNG/30 EE. Conclusions: DRSP/20 EE and LNG/30 EE induce favorable changes in endothelial function. (C) 2012 Elsevier Inc. All rights reserved.

Seasons of desiccant application in the soybean crop: water content and productivity

The herbicides glyphosate and paraquat have been used by Brazilian soybean producers to obtain crop desiccation and to anticipate and uniformity at harvest. However, improper use of herbicides can to occasion problems in agronomic and physiologic characteristics of crop. This study aimed to evaluate the use of the glyphosate and paraquat herbicides as a desiccant for growing soybeans. The experiment was conducted in 2005/06 crop year, in an experimental design of randomized blocks with four replications. Treatments were arranged in two factorial design, 3x3x5x2 and 3x2x5x2: two desiccants (glyphosate and paraquat) and control (without drying), three and two growth stages (R6, R7 and R8) and (R7 and R8) for varieties MSOY 6101 of superprecocious cycle, and MG / BR 46 (Conquista) of precocious cycle, respectively, five sampling times (2, 4, 6, 8 and 10 days after application) and two positions in the plants on the ground assessment (apex and base). Was evaluated for seed Production, mass and water content of 100 seeds. Desiccants tested were effective in reducing the water content of seeds, without affecting productivity and mass of 100 seeds.

Seasonal Distribution of Airborne Trace Elements and Water-Soluble Ions in Sao Paulo Megacity, Brazil

This study deals with the seasonal distribution of Al, Ca, Cu, Fe, K, Mg, Na, Pb and Zn and water soluble ions (Cl-, PO43-, NO3-, SO42-, HCOO-, CH3COO-, oxalate, succinate, Na+, NH4+, K+, Mg2+ and Ca2+) found in PM10 samples (particulate matter less than 10 mu m in diameter) Sao Paulo City, Brazil, (April 2003-May 2004). Higher atmospheric levels were found for SO42-, NO3-, Cl- and PO43- while the main organic anions were oxalate and formate. Atmospheric levels for elements were: Fe > Al > Ca > K > Na > Mg > Zn > Cu > Pb. Some sources were predominant for some species: (i) fuel burning and/or biomass burning (NO3-, HCOO-, C2O42-, K+, Mg2+, Ca2+, Fe, Pb, Zn, Al, Ca, K and Mg), (ii) gas-to-particle conversion (SO42- and NH4+) and (iii) sea salt spray (Cl-, Na+ and Na).

Using Fluid Bed Granulation to Improve the Dissolution of Poorly Water-Soluble Drugs

In this study, fluid bed granulation was applied to improve the dissolution of nimodipine and spironolactone, two very poorly water-soluble drugs. Granules were obtained with different amounts of sodium dodecyl sulfate and croscarmellose sodium and then compressed into tablets. The dissolution behavior of the tablets was studied by comparing their dissolution profiles and dissolution efficiency with those obtained from physical mixtures of the drug and excipients subjected to similar conditions. Statistical analysis of the results demonstrated that the fluid bed granulation process improves the dissolution efficiency of both nimodipine and spironolactone tablets. The addition of either the surfactant or the disintegrant employed in the study proved to have a lower impact on this improvement in dissolution than the fluid bed granulation process.