Elemental analysis of Brazilian coffee with ion beam techniques: From ground coffee to the final beverage.

Brazilian coffee is well known worldwide due to its quality and richness in taste. The aim of the present study is to provide the elemental characterization of Brazilian coffee along different stages of the drip brewing process. To that end, samples from roasted ground coffee, spent coffee, paper filters and the final beverage were analyzed with one single ion beam technique, namely particle-induced X-ray emission (PIXE). In total, over 140 samples from 8 different Brazilian brands of ground coffee were analyzed. Large differences in some elemental concentrations were observed among different brands and among different batches of a single brand, which leads to high variances in the data. Concerning the beverage preparation, the analysis of the spent coffee shows that the transfer ratio from the ground coffee to the beverage differs for each element. Our results indicate that potassium and chlorine have the highest transfer ratio. Moreover, the concentration of rubidium is relatively high in drinking coffee. Finally, there is no influence of the elemental composition of paper filter in the preparation of drinking coffee.

Enhanced resistance of yeast mutants deficient in low-affinity iron and zinc transporters to stannous-induced toxicity.

Tin or stannous (Sn(2+)) compounds are used as catalysts, stabilizers in plastic industries, wood preservatives, agricultural biocides and nuclear medicine. In order to verify the Sn(2+) up-take and toxicity in yeast cells we utilized a multi-elemental analysis known as particle-induced X-ray emission (PIXE) along with cell survival assays and quantitative real-time PCR. The detection of Sn(2+) by PIXE was possible only in yeast cells in stationary phase of growth (STAT cells) that survive at 25mM Sn(2+) concentration. Yeast cells in exponential phase of growth (LOG cells) tolerate only micro-molar Sn(2+) concentrations that result in intracellular concentration below of the method detection limit. Our PIXE analysis showed that STAT XV185-14c yeast cells demonstrate a significant loss of intracellular elements such as Mg, Zn, S, Fe and an increase in P levels after 1h exposure to SnCl(2). The survival assay showed enhanced tolerance of LOG yeast cells lacking the low-affinity iron and zinc transporters to stannous treatment, suggesting the possible involvement in Sn(2+) uptake. Moreover, our qRT-PCR data showed that Sn(2+) treatment could generate reactive oxygen species as it induces activation of many stress-response genes, including SOD1, YAP1, and APN1.

Antioxidant activity and phenolic and mineral content of rose grape juice.

There are many studies related to the antioxidant activity of grape products; however, they concern only purple and white grape varieties. Up to now, there are no reports of studies on the Goethe rose grape variety, either on its antioxidant activity or on its phenolic and mineral quantification. Thus, the purpose of this study was to evaluate in vitro and in vivo antioxidant activity, as well as to quantify total phenolic compounds, ascorbic acid, and mineral content, in a Goethe rose grape juice. The results obtained showed that the Goethe rose grape juice is a great polyphenol source, which contains catechin, epicatechin, and procyanidins (B(1), B(2), B(3), and B(4)). Of all metals analyzed, potassium, calcium, magnesium, and iron showed the highest values. We found that this rose grape juice shows an important antioxidant activity in in vitro (2,2-diphenyl-1-picrylhydrazyl radical scavenging activity) and in vivo (using the Saccharomyces cerevisiae yeast cells) assays. The antioxidant activity could be explained by the significant phenolic content and ascorbic acid levels found in the juice. The results showed that rose grape juice is an excellent antioxidant source, which could contribute to the prevention of many diseases related to oxidative stress, such as atherosclerosis and Parkinson's disease.

The Pmr1 protein, the major yeast Ca2+-ATPase in the Golgi, regulates intracellular levels of the cadmium ion.

Cadmium is a nonessential, highly toxic heavy metal that shows ionic properties similar to calcium. These ionic similarities imply that the cadmium ion, Cd2+, is a calcium ion, Ca2+, receptor-agonist, affecting the same biochemical pathways involved in Ca2+ homeostasis. In the yeast Saccharomyces cerevisiae, the PMC1 and PMR1 genes encode vacuolar and Golgi Ca2+-ATPases, respectively. The PMR1 protein product Pmr1p is involved in both Ca2+ and Mn2+ homeostasis. This study investigated the importance of Pmc1p and Pmr1p for Cd2+ cellular detoxification. Using the standard techniques of yeast molecular research and a multielemental procedure named particle-induced X-ray emission, Pmr1p was identified as a protein that directly participates in the detoxification of Cd2+, possibly through the secretory pathway. The results allow us to posit a model of Cd2+ detoxification where Pmr1p has a central role in cell survival in a Cd2+-rich environment.

Genotoxicity and mutagenicity of iron and copper in mice.

The toxicity of trace metals is still incompletely understood. We have previously shown that a single oral dose of iron or copper induces genotoxic effects in mice in vivo, as detected by single cell gel electrophoresis (comet assay). Here, we report the effect of these metals on subchronic exposure. Mice were gavaged for six consecutive days with either water, 33.2 mg/kg iron, or 8.5 mg/kg copper. On the 7th day, the neutral and alkaline comet assays in whole blood and the bone marrow micronucleus (MN) test were used as genotoxicity and mutagenicity endpoints, respectively. Particle induced X-ray emission was used to determine liver levels of the metals. Females showed a slightly lower DNA damage background, but there was no significant difference between genders for any endpoint. Iron and copper were genotoxic and mutagenic. While copper was more genotoxic in the neutral version, iron was more genotoxic in the alkaline version of the comet assay. Copper induced the highest mutagenicity as evaluated by the MN test. Iron was not mutagenic to male mice. Iron is thought to induce more oxidative lesions than copper, which are primarily detected in the alkaline comet assay. Treatment with iron, but not with copper, induced a significant increase in the hepatic level of the respective metal, reflecting different excretion strategies.

Elemental characterization of commercial mate tea leaves (Ilex paraguariensis A. St.-Hil.) before and after hot water infusion using ion beam techniques.

Ilex paraguariensis A. St.-Hil. is used to prepare a traditional tealike beverage widely appreciated in Argentina, Paraguay, Uruguay, and southern Brazil. In these countries, the tea is popularly known as mate or chimarrão. The aim of this work is to characterize the elemental composition of commercial Ilex paraguariensis and determine the portion of each element present in the leaves that is eluted in the water during the infusion process and consequently ingested by the drinker. Using the particle-induced X-ray emission technique, we verified the presence of Mg, Al, Si, P, S, Cl, K, Ca, Ti, Mn, Fe, Cu, Zn, and Rb at different concentrations, which accounts for about 3.4% of the total mass. The results show a loss of about 90% of K and Cl, 50% of Mg and P, and 20% of Mn, Fe, Cu, Zn, and Rb by the leaves after the infusion. The volume of water used in the infusion affects only the concentration of elements such as Cl, P, K, and Mg until the first 600 mL of water, where a steep decrease in the concentration of these elements was observed in brewed leaves. Furthermore, higher water temperatures (typical temperatures used in infusions, between 80 and 100 degrees C) favor the extraction of K and Cl into the infusion, while the concentration of other elements remains practically constant as a function of temperature.

Influence of orange juice in the levels and in the genotoxicity of iron and copper.

World consumption of natural juices is increasing as a consequence of the human search for a healthier life. The juice production industry, especially for orange juice, is expanding in several countries and particularly in Brazil. Despite scientific data reporting beneficial properties derived from juice consumption, some components of juices have been identified as mutagenic or carcinogenic. Carcinogenic or genotoxic effects may be mediated by the interaction of juice components with transition metals or by sub-products of juice auto-oxidation. In this study, the mutagenic potential of orange juice and two metallic agents used in dietary supplementation, FeSO(4) and CuSO(4), were investigated using the comet assay in mouse blood cells (in vivo). Both metal compounds were genotoxic for eukaryotic cells after 24h treatment at the doses used. Significant damage repair was observed after 48h of treatment with the same compounds. Orange juice had a modulating effect on the action of metallic sulfates. In the case of iron treatment, the presence of the orange juice had a preventive, but not restorative, effect. On the other hand, in the case of copper treatment, the effects were both preventive and restorative. PIXE (particle induced X-ray emission) analysis indicated a positive correlation between DNA damage and the hepatic levels of iron and a negative correlation between whole blood copper and DNA damage. A negative correlation between hepatic iron and whole blood copper content was also seen in the treatment with both ferrous and cupric sulfates.

The function of Alr1p of Saccharomyces cerevisiae in cadmium detoxification: insights from phylogenetic studies and particle-induced X-ray emission.

Two genes in Saccharomyces cerevisiae, ALR1 and ALR2, encode transmembrane proteins involved in Mg2+ uptake. The present study investigates the phylogenetic relationship of Alr1p/Alr2p with bacterial CorA proteins and some proteins related to Mg2+ influx/efflux transport in mitochondrial and bacterial zinc transporters; including hydrophobic cluster analysis (HCA). The phylogenetic results indicate that the Alrp sequences of S. cerevisiae share a common carboxy-terminus with proteins related to zinc efflux transport. We also analyse the intracellular metal content by particle-induced X-ray emission (PIXE) after cell exposure to cadmium. The PIXE analysis of cadmium-exposed ALR mutants and wild-type yeast cells suggests that Alrp has a central role in cell survival in a cadmium-rich environment.

Fish as bioindicators to assess the effects of pollution in two southern Brazilian rivers using the Comet assay and micronucleus test.

Industrial effluents, agricultural runoff, and municipal wastewaters contain unknown substances and complex mixtures that are released into the environment and can lead to contamination of surface and subsurface waters. In the present report, we have used the alkaline Comet assay and the micronucleus (MN) test to detect the genotoxicity due to multiple sources of pollution in the peripheral blood of two native estuarine fish (mullet and sea catfish) and evaluated possible interactive genotoxic effects from multiple contaminants and the seasonal variation of the genotoxicity. Mullet and sea catfish were captured in the Tramandai and Mampituba Rivers in the southern Brazilian state of Rio Grande do Sul. Reference animals were obtained from the Armazem lagoon. Fish captured in the two estuaries during the four seasons over a period of 2 years had increased levels of DNA damage and MN frequencies relative to the reference fish. In general, the alkaline Comet assay was more sensitive to the genotoxicity of the river contaminants than the MN test. The Comet assay demonstrated significant differences in fish captured at different seasons and at the two river sites, while the MN test showed significant differences only for the annual average for mullet from both sites and fish from the control site. The increases in DNA damage appear to be related to the increase in the number of people in the towns close to the study areas during the warm spring and summer seasons. Although no specific cause-effect relationships were established, comparison of the chemical contaminants and physical variations in the rivers with the genotoxicity data indicate that there may be some association between hydrocarbons, metals, pH, and water temperature and the level of damaged cells observed in mullet and sea catfish from the Tramandai and Mampituba estuaries.

Genotoxicity biomonitoring in regions exposed to vehicle emissions using the comet assay and the micronucleus test in native rodent Ctenomys minutus.

Exposure to motor vehicle emissions represents an important concern for possible long-term health effects. The present report describes: 1) the application and verification of the alkaline comet assay in Ctenomys minutus to detect the possible genotoxicity of automobile emissions; 2) a comparison of the comet assay results with peripheral blood micronucleus (MN) assay results performed in the same animals; and 3) the identification of agents involved in the responses and in the seasonal variation of the effects. Ctenomys minutus (Octodontidae-Rodentia) were captured in two different fields from both sides of RS/030, a highway on the coastal plain of the Brazilian state of Rio Grande do Sul. Reference animals were obtained from a nearby field that was about 3 km distant from any road. By the end of this study, 123 rodents (73 females and 50 males) were live-trapped. Our results indicate that there was an increase in cells with DNA damage for C. minutus environmentally exposed to automobile emissions, as demonstrated by the alkaline comet assay, but there was no increase in micronucleated cells. The alkaline comet assay showed age and gender differences in the response. The comet assay results suggest that adult females are the principal population affected by air pollutants from vehicle emissions. Chemical data were also collected from areas exposed to automobile exhaust and these indicated that elevated levels of hydrocarbons, metals, and NO(2) were associated with the elevated levels of damaged cells observed in the wild rodent C. minutus. Our results agree with previous data on engine and fuel components, where weak increases in damage for native rodents exposed to emissions have been observed. Other larger, controlled studies are needed to better understand how the metabolism of C. minutus affects its response to emission exposure.