Comparison of Analytical Methods for Quantifying the Levels of Lead,Cadmium, Chromium,and Nickel in Mineral Fertilizers and Industrial By-products

The control of contaminants in fertilizers and industrial by-products is fundamental to safe food production, and it requires low-cost, reliable test methods. The aim of this study was to evaluate the performance of the USEPA 3051a and alternative analytical methods [concentrated hydrochloric acid (HCl), 10 percent HCl, 10 percent sulfuric acid (H₂SO₄), and 50 mmol L^?1 diethylenetriaminepentaacetic acid (DTPA)] for quantifying the levels of lead, cadmium, chromium, and nickel in zinc and/or copper sources. The sample digestions with concentrated HCl, H₂SO₄, and nitric acid (HNO₃) (USEPA 3051a) were heated. Extraction without heating and stirring was employed in 10 percent HCl and 50 mmol L^?1 DTPA methods. The Graybill’s modified F-test, t-test for mean error, and the linear correlation coefficient analysis were used to compare test method performance. Equivalent results to USEPA 3051a were found with the following methods: DTPA for the extraction of lead in zinc sources and concentrated HCl and H₂SO₄ for the extraction of cadmium and chromium in copper sources. However, the absolute values of cadmium, chromium, and nickel recovered by the use of H₂SO₄ were greater regardless of the source. The greater extraction of contaminants in the evaluated methods suggests that there is need to review the official method if the aim is to quantify the total levels of these elements in raw materials and mineral fertilizers.     

Importance of heat shock proteins in maize

Abiotic and biotic stress conditions cause extensive losses to maize production, mainly due to protein dysfunction in these conditions. In higher plants, the occurrence of heat-shock proteins (HSPs) in response to different environmental stresses is a universal phenomenon and has been well documented. Many studies have demonstrated that most HSPs are involved in many regulatory pathways, act as molecular chaperones for other cell proteins, and have strong cytoprotective effects. Although many functional roles for HSPs are known, the mechanisms for these multiple functions are not entirely understood. Here we reviewed the correlation among HSP genes/proteins and plant tolerance, especially maize, in different environmental stresses. Due to the low availability of information regarding the expression of HSP genes in response to different stresses in maize, we decided to mine databases in order to generate new insights related to this topic.     

In situ characterization of the granulomatous immune response with time in nonhealing lesional skin of Leishmania braziliensis-infected rhesus macaques (Macaca mulatta)

We have recently introduced a macaque (Macaca mulatta) model of Leishmania braziliensis-induced self-healing cutaneous leishmaniasis in which the T cell-mediated inflammatory response effectively promotes parasite clearance and granuloma resolution. Here we show that macaques infected with a highly pathogenic L. braziliensis strain displayed longstanding granulomatous lesions which lasted until the end of the observation period (52 weeks). Immunoperoxidase staining of representative tissue sections indicated that distinct cell populations (CD3, CD4, CD8, CD20, Foxp3, CD20, CD68, HLA-DR, CCL2, and CXCL-10) change uniformly during infection, suggesting that the same components of the local immune response are working in unison. This model also confirmed that granuloma formation is orchestrated by diverse inflammatory mediators that are important for T helper type 1 (Th1) cell development and macrophage effector functions. Cytometry analysis of ex vivo granuloma-derived leukocytes revealed accumulation of distinct functional subsets of effector and regulatory T cells into the inflamed skin. We provide evidence that local interleukin (IL)-10 production by both Foxp3(+) and Foxp3(-) CD4(+) T subsets is likely important in promoting lesional granuloma maintenance. Further studying the immune suppression mechanisms that induces granulomas in L. braziliensis-infected macaques may reveal new opportunities for therapeutic control of this important human disease.     

In vitro effect of Aloe vera, Coriandrum sativum and Ricinus communis fractions on Leishmania infantum and on murine monocytic cells

In South America, visceral leishmaniasis is a zoonosis caused by the protozoan species Leishmania infantum (syn. L. chagasi) and is primarily transmitted through the bite of the female Lutzomyia longipalpis. Its main reservoir in urban areas is the dog. The application of control measures recommended by health agencies have not achieved significant results in reducing the incidence of human cases, and the lack of effective drugs to treat dogs resulted in the prohibition of this course of action in Brazil. Therefore, it is necessary to search new alternatives for the treatment of canine and human visceral leishmaniasis. The objectives of this study were to evaluate the in vitro effect of fractions from Aloe vera (aloe), Coriandrum sativum (coriander), and Ricinus communis (castor) on promastigotes and amastigotes of L. infantum and to analyze the toxicity against the murine monocytic cells RAW 264.7. To determine the viability of these substances on 50% parasites (IC50), we used a tetrazolium dye (MTT) colorimetric assay (bromide 3-4.5-dimethylthiazol-2-yl-2,5-dephenyltetrazolium), and on amastigotes we performed an in situ ELISA. All fractions were effective against L. infantum promastigotes and did not differ from the positive control pentamidine (p>0.05). However, the R. communis ethyl acetate and chloroform fractions, as well as the C. sativum methanol fraction, were the most effective against amastigotes and did not differ from the positive control amphotericin B (p>0.05). The R. communis ethyl acetate fraction was the least toxic, presenting 83.5% viability of RAW 264.7 cells, which was similar to the results obtained with amphotericin B (p>0.05). Based on these results, we intend to undertake in vivo studies with R. communis ethyl acetate fractions due the high effectiveness against amastigotes and promastigotes of L. infantum and the low cytotoxicity towards murine monocytic cells.     

Influence of category-heifers,primiparous and multiparous lactating cows-in a large-scale resynchronization fixed-time artificial insemination program

This study was conducted to evaluate the influence of category (heifers, primiparous or multiparous cows) on pregnancy rates in a large scale resynchronization ovulation program. Nelore heifers (n = 903), primiparous lactating cows (n = 338) and multiparous lactating cows (n = 1,223) were synchronized using a conventional protocol of estradiol/P4-based fixed-time artificial insemination (FTAI). Thirty days after ultrasonography, females who failed the first FTAI were resynchronized with the same hormonal protocol prior to a second FTAI. The pregnancy status of each cohort was evaluated by ultrasonography 30 days after each FTAI. The average conception rate after the first FTAI and resynchronization was 80.5%. Heifers had a higher conception rate (85%) than primiparous (76%) or multiparous cows (78%; p = 0.0001). The conception rate after the first FTAI was similar among heifers (57%), primiparous cows (51%) and multiparous cows (56%; p = 0.193). After the second FTAI, heifers exhibited a higher conception rate (66%) than primiparous or multiparous cows (51%; p = 0.0001). These results demonstrate the feasibility of resynchronization in large beef herds for providing consistent pregnancy rates in a short period of time. We also demonstrated that ovulation resynchronization 30 days after FTAI is particularly effective for heifers, providing a conception rate of up to 66%.     

Effects of bovine leukemia virus infection on milk neutrophil function and the milk lymphocyte profile

The effects of bovine leukemia virus (BLV) on the immune response have been extensively investigated; however, its effects on mammary gland immunity are only speculative. Although BLV has a tropism for B cells, it can affect both adaptive and innate immunities because these systems share many effector mechanisms. This scenario is the basis of this investigation of the effects of BLV on mammary gland immunity, which is largely dependent upon neutrophilic functions. Thus, the present study sought to examine neutrophilic functions and the lymphocyte profile in the milk of naturally BLV-infected cows. The viability of the milk neutrophils and the percentage of milk neutrophils that produced reactive oxygen species (ROS) or phagocytosed Staphylococcus aureus were similar between BLV-infected and BLV-uninfected dairy cows. Furthermore, the expression of CD62L and CD11b by the milk neutrophils and the percentage of milk neutrophils (CH138⁺ cells) that were obtained from the udder quarters of the BLV-infected cows were not altered. Conversely, the median fluorescence intensity (MFI) representing intracellular ROS production and the phagocytosis of S. aureus, the expression of CD44 by the milk neutrophils and the percentage of apoptotic B cells were lower in the milk cells from BLV-infected dairy cows, particularly those from animals with persistent lymphocytosis (PL). The lymphocyte subsets were not different among the groups, with the exception of the percentage of CD5⁻/CD11b⁻ B cells, which was higher in the milk cells from BLV-infected cows, particularly those with PL. Thus, the present study provides novel insight into the implications of BLV infection for mammary gland immunity.     

New Insights of Turmeric Extract-Loaded PLGA Nanoparticles: Development,Characterization and In Vitro Evaluation of Antioxidant Activity

Here, we presented new insights of the development of poly(lactic-co-glycolic acid) nanoparticles containing turmeric compounds (turmeric-PLGA-NPs) using emulsion-solvent evaporation method. The nanoparticulate system was characterized by size, zeta potential, morphology, release profile, partition parameter, stability and encapsulation efficiency (%EE). Antioxidant activity studies were also evaluated. The Korsmeyer-Peppas model (M_t/M_∞ vs. t) was used to determine the release mechanisms of the studied system. Our results demonstrated the emulsion-solvent evaporation method was shown advantageous for producing turmeric-PLGA-NPs in the range of 145 nm with high homogeneity in size distribution, zeta potential of ?21.8 mV and %EE about 72%. Nanoparticles were stable over a period of one month. In vitro study showed a release of curcumin governed by diffusion and relaxation of the polymeric matrix. The partition parameter of the extract in relation to blank-PLGA-NPs was 0.111?±?0.008 M^?1, indicating a low affinity of curcumin for the polymer matrix. Antioxidant ability of the turmeric-PLGA-NPs in scavenging the radical 2,2-azinobis (3-ethylbenzothiazoline- 6-sulfonic acid) (ABTS) was inferior to free turmeric extract and showed a concentration and time-dependent profile. The study concluded that PLGA nanoparticles are potential carriers for turmeric extract delivery.

     

Vasorelaxation, induced by Dictyota pulchella (Dictyotaceae), a brown alga, is mediated via inhibition of calcium influx in rats

This study aimed to investigate the cardiovascular effects elicited by Dictyota pulchella, a brown alga, using in vivo and in vitro approaches. In normotensive conscious rats, CH(2)Cl(2)/MeOH Extract (CME, 5, 10, 20 and 40 mg/kg) from Dictyota pulchella produced dose-dependent hypotension (-4 ± 1; -8 ± 2; -53 ± 8 and -63 ± 3 mmHg) and bradycardia (-8 ± 6; -17 ± 11; -257 ± 36 and -285 ± 27 b.p.m.). In addition, CME and Hexane/EtOAc Phase (HEP) (0.01-300 μg/mL) from Dictyota pulchella induced a concentration-dependent relaxation in phenylephrine (Phe, 1 μM)-pre-contracted mesenteric artery rings. The vasorelaxant effect was not modified by the removal of the vascular endothelium or pre-incubation with KCl (20 mM), tetraethylammonium (TEA, 3 mM) or tromboxane A(2) agonist U-46619 (100 nM). Furthermore, CME and HEP reversed CaCl(2)-induced vascular contractions. These results suggest that both CME and HEP act on the voltage-operated calcium channel in order to produce vasorelaxation. In addition, CME induced vasodilatation after the vessels have been pre-contracted with L-type Ca(2+) channel agonist (Bay K 8644, 200 nM). Taken together, our data show that CME induces hypotension and bradycardia in vivo and that both CME and HEP induce endothelium-independent vasodilatation in vitro that seems to involve the inhibition of the Ca(2+) influx through blockade of voltage-operated calcium channels.     

Evaluation of antiradical capacity by H2O2-hemin-induced luminol chemiluminescence

This paper describes a screening method for antioxidant potential determination based on luminol/hemin/hydrogen peroxide chemiluminescence. The emission depletion, caused by an antiradical compound added during the chemiluminescence decay, is proportional to the number of reactive species trapped. Therefore, the difference between the areas of the emission decay curves, obtained in the absence and in the presence of the potential antioxidant, is a measure for the antiradical capacity of the sample. The technique has been applied to measure the antiradical capacity of pure compounds and complex mixtures from natural origin, providing reliable results that indicate the method's feasibility.     

Cross-amplification and characterization of microsatellite loci for three species of <Emphasis Type="Italic">Theobroma</Emphasis> (Sterculiaceae) from the Brazilian Amazon

This study reports on the cross-species amplification of 23 microsatellite markers previously developed for Theobroma cacao L. (Sterculiaceae), source of chocolate in three economically important Amazonian species of Theobroma (T. grandiflorum, T. subincanum, T. sylvestre). Thirteen of the 23 microsatellite loci tested were polymorphic across the three species at 2–13 alleles per locus. The observed heterozygosity per locus varied from 0.18 to 0.84 and expected heterozygosity ranged from 0.28 to 0.87. The high level of transferability and genetic information content of these microsatellite loci indicate their usefulness for population genetic, mating system and breeding studies of these economically important Amazonian fruit trees.