A simple and automated sample preparation system for subsequent halogens determination: Combustion followed by pyrohydrolysis.

A simple and automated system based on combustion followed by a pyrohydrolysis reaction was proposed for further halogens determination. This system was applied for digestion of soils containing high (90%) and also low (10%) organic matter content for further halogens determination. The following parameters were evaluated: sample mass, use of microcrystalline cellulose and heating time. For analytes absorption, a diluted alkaline solution (6 mL of 25 mmol L-1 NH4OH) was used in all experiments. Up to 400 mg of soil with high organic matter content and 100 mg of soil with low organic matter content (mixed with 400 mg of cellulose) could be completely digested using the proposed system. Quantitative results for all halogens were obtained using less than 12 min of sample preparation step (about 1.8 min for sample combustion and 10 min for pyrohydrolysis). The accuracy was evaluated using a certified reference material of coal and spiked samples. No statistical difference was observed between the certified values and results obtained by the proposed method. Additionally, the recoveries obtained using spiked samples were in the range of 98-103% with relative standard deviation values lower than 5%. The limits of quantification obtained for F, Cl, Br and I for soil with high (400 mg of soil) and low (100 mg of soil) organic matter were in the range of 0.01-2 µg g-1 and 0.07-59 µg g-1, respectively. The proposed system was considered as a simple and suitable alternative for soils digestion for further halogens determination by ion chromatography and inductively coupled plasma mass spectrometry techniques.

Efeito metafilático do zinco injetável sobre parâmetros metabólicos e oxidativos de ovelhas no pós-parto imediato / Metaphylactic effect of injectable zinc on metabolic and oxidative parameters of sheep in the immediate postpartum

The aim of this study was to evaluate a new source of injectable organic zinc (zinc edetate) on the energy and oxidative profile in sheep during the immediate postpartum period. Twenty-six Texel breed animals were previously identified and divided into two experimental groups: the treated group (TG; n= 13) that comprised the animals that received a subcutaneous (SC) injection of 100 mg of zinc edetate (2 mL) fifteen days before the parturition expected date and the control group (CG; n=13) that comprised the animals that received 2mL of physiological solution at the same date of TG. Blood samples were collected on the parturition day for the assessment of serum fructosamine, cholesterol and triglycerides, insulin-like growth factor type 1 (IGF-1), the oxidative stress index (OSi) and blood zinc concentration. In addition to these parameters, the measurement of zinc was made in food given to the animals. There was no difference in metabolic parameters and OSi between the experimental groups (P>0.05), as well as in blood zinc concentrations (P>0.05). The parenteral zinc edentate does not change the energy and oxidative profile of sheep in immediate postpartum.(AU)

Multitechnique Determination of Halogens in Soil after Selective Volatilization Using Microwave-Induced Combustion.

A method for digestion of soils with high inorganic matter content (ranging from 50 to 92%) by microwave-induced combustion (MIC) is proposed for the first time for further halogens (F, Cl, Br, and I) determination by ion chromatography (IC) and also by inductively coupled plasma mass spectrometry (ICP-MS). Microcrystalline cellulose (100-500 mg), used as a combustion aid, was mixed with sample and water or NH4OH solutions (10-100 mmol L-1) were investigated for analytes absorption. The use of cellulose (400 mg) was mandatory to volatilize the halogens from soils with high inorganic matter. It was possible to use diluted absorbing solutions (up to 100 mmol L-1 NH4OH) for halogens retention, providing limits of quantification in the range of 0.06 (I) to 60 (Cl) µg g-1. Accuracy was evaluated using certified reference materials (CRMs), spiked samples, and pyrohydrolysis method. Recoveries for halogens after spiked samples were in the range of 94 to 103% and the results after digestion of CRMs by MIC were in agreement better than 95% to certified values. Blanks were low, relative standard deviation was below 8% for all soils and no statistical difference was observed for results by pyrohydrolysis and MIC methods showing the feasibility of the proposed method for further halogens determination in soil samples.

Association between bone turnover markers, clinical variables, spinal syndesmophytes and bone mineral density in Mexican patients with ankylosing spondylitis.

OBJECTIVES: To compare bone turnover marker (BTM) levels and bone mineral density (BMD) between patients with ankylosing spondylitis (AS) and healthy controls (HC) and to evaluate, in AS, the association between BTM levels and clinical variables, spinal syndesmophytes, and BMD using multivariate analysis. METHOD: Seventy-eight AS patients were compared with 58 HC matched by gender. Spinal syndesmophytes in AS and other characteristics were assessed. C-terminal telopeptide fragments of type I collagen (CTX), bone-specific alkaline phosphatase (BAP), osteocalcin (OC) serum levels, and BMD of the lumbar spine, femoral neck, and forearm were evaluated. RESULTS: AS males and females had lower BAP levels than their respective HC (p < 0.001 and p = 0.001). AS patients with bridging syndesmophytes had higher OC levels than AS patients either with non-bridging syndesmophytes (p = 0.001) or without spinal syndesmophytes (p < 0.001). OC and CTX levels correlated significantly with the modified Stoke Ankylosing Spondylitis Spinal Score (mSASSS). In the multivariate linear regression adjusted by age, gender, the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), BMD in the lumbar spine, and C-reactive protein (CRP), we observed an association between BAP levels and anti-tumour necrosis factor (anti-TNF) use (p = 0.05) whereas OC levels were associated with mSASSS (p < 0.001) and anti-TNF use (p = 0.05), and CTX levels were exclusively associated with mSASSS (p = 0.03). In the logistic regression analysis, only OC levels were associated with the presence of syndesmophytes in AS [odds ratio (OR) 2.42, 95% confidence interval (CI) 1.19-5.75]. CONCLUSIONS: We observed an increase in OC levels in AS patients with syndesmophytes. BTM levels were associated with the severity of spinal damage. Future longitudinal studies should evaluate whether these BTMs should be included as tools to determine the prognosis and progression of spinal damage.

Microwave-induced combustion of crude oil for further rare earth elements determination by USN-ICP-MS.

A procedure for light and heavy crude oils digestion by microwave-induced combustion (MIC) is proposed for the first time for further rare earth elements (REE) determination by inductively coupled plasma mass spectrometry (ICP-MS) equipped with an ultrasonic nebulizer (USN). Samples of crude oil (API density of 10.8-23.5, up to 250 mg) were inserted in polycarbonate capsules and combusted using 20 bar of oxygen and 50 µL of 6 mol L(-1) ammonium nitrate as igniter. Nitric acid solutions (1-14.4 mol L(-1)) were evaluated for analyte absorption and a reflux step was applied after combustion (5 min of microwave irradiation at 1400 W) in order to achieve better analyte recoveries. Accuracy was evaluated using a spiked sample and also by comparison of results obtained by microwave-assisted digestion combined to ultraviolet radiation (MW-UV) and by neutron activation analysis (NAA). Using 3 mol L(-1) HNO3, quantitative recoveries (better than 97%) were obtained for all analytes. Blank values were always negligible. Agreement was higher than 96% for La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu and Y by comparison of results with those obtained by MW-UV and by NAA (only for La, Ce, Nd, Sm, and Yb). Residual carbon content in digests using MIC was always below 1%. As an advantage over conventional procedures for crude oil digestion, using MIC, it was possible to use diluted acid as absorbing solution, obtaining better limits of detection and avoiding interferences in REE determination by USN-ICP-MS.

Effects of Hg(II) exposure on MAPK phosphorylation and antioxidant system in D. melanogaster.

The heavy metal mercury is a known toxin, but while the mechanisms involved in mercury toxicity have been well demonstrated in vertebrates, little is known about toxicological effects of this metal in invertebrates. Here, we present the results of our study investigating the effects associated with exposure of fruit fly Drosophila melanogaster to inorganic mercury (HgCl2 ). We quantify survival and locomotor performance as well as a variety of biochemical parameters including antioxidant status, MAPK phosphorylation and gene expression following mercury treatment. Our results demonstrate that exposure to Hg(II) through diet induced mortality and affected locomotor performance as evaluated by negative geotaxis, in D. melanogaster. We also saw a significant impact on the antioxidant system including an inhibition of acetylcholinesterase (Ache), glutathione S-transferase (GST) and superoxide dismutase (SOD) activities. We found no significant alteration in the levels of mRNA of antioxidant enzymes or NRF-2 transcriptional factor, but did detect a significant up regulation of the HSP83 gene. Mercury exposure also induced the phosphorylation of JNK and ERK, without altering p38(MAPK) and the concentration of these kinases. In parallel, Hg(II) induced PARP cleavage in a 89 kDa fragment, suggesting the triggering of apoptotic cell death in response to the treatment. Taken together, this data clarifies and extends our understanding of the molecular mechanisms mediating Hg(II) toxicity in an invertebrate model.

High-efficiency microwave-assisted digestion combined to in situ ultraviolet radiation for the determination of rare earth elements by ultrasonic nebulization ICPMS in crude oils.

A method for heavy and extraheavy crude oil digestion based on microwave-assisted wet digestion (MW-AD) and ultraviolet (UV) radiation using diluted HNO3 was applied for the determination of rare earth elements (REE) by inductively coupled plasma mass spectrometry (ICPMS) with an ultrasonic nebulizer (USN). Even using pressurized systems conventional acid digestion is not feasible for efficient crude oil digestion, especially for heavy and extraheavy crude oils that generally present high amounts of asphaltenes and resins. In the proposed system, UV radiation is generated in situ by immersed electrodeless Cd discharge lamps positioned inside quartz vessels. The use of diluted solutions (1-14.4 mol L(-1) HNO3 and 1-4 mol L(-1) H2O2) were evaluated for heavy and extraheavy crude oil digestion (API density of 11.1-19.0). With the proposed method the residual carbon content was lower than 13 mg C/100 mg of sample, and it was possible to digest sample masses up to 500 mg using 4 mol L(-1) HNO3 and 4 mol L(-1) H2O2. Interferences caused by excessive acid concentration and carbon content in digests were minimized allowing limits of quantification for REEs as low as 0.3 ng g(-1). Samples were also digested using MW-AD in pressurized systems with concentrated HNO3, but even using 280 °C, 80 bar, and concentrated HNO3, MW-AD method was not suitable for REE determination due to interferences in ICPMS determination. The combination of microwave heating with UV was considered a suitable and effective way to digest crude oil allowing further determination of low concentrations of REE by ICPMS.

Moderate hypoxia is able to minimize the manganese-induced toxicity in tissues of silver catfish (Rhamdia quelen).

The aim of this study was to compare the effects of manganese (Mn) on silver catfish exposed to different levels of dissolved oxygen. Silver catfish (Rhamdia quelen) were exposed to increasing concentrations of Mn (4.2, 8.4 or 16.2mgL(-1)) under either normoxia (100 percent saturation) or moderate hypoxia (51.87 percent saturation) for 15 days. Under normoxia, Mn exposure increased lipid peroxidation (LP) in brain and kidney; it increased gluthatione (GSH) levels in brain and decreased catalase (CAT) activity in both tissues. Moderate hypoxia was able to prevent Mn-induced LP in brain and to reduce this oxidative parameter in kidney; GSH level was increased in brain, while CAT activity was reduced in both tissues. Activity of isolated mitochondria of liver and gills was reduced by Mn exposure under both levels of dissolved oxygen, but this effect was more prominent in normoxia. As expected, liver, kidney and gills showed an increase of Mn accumulation according to waterborne levels, and these parameters presented positive relationship. The highest waterborne Mn (8.4 and 16.2mgL(-1)) resulted in greater accumulation under normoxia, indicating that moderate hypoxia can stimulate mechanisms capable of reducing Mn accumulation in tissues (though not in blood). Moderate hypoxia can be considered a stress factor and Mn an aquatic anthropogenic contaminant. Therefore we hypothesized that these two conditions together are able to invoke defense mechanisms in juvenile silver catfish, acting in a compensatory form, which may be related to adaptation and/or hormesis.

Prescription for antiresorptive therapy in Mexican patients with rheumatoid arthritis: is it time to reevaluate the strategies for osteoporosis prevention?

Glucocorticoids are frequently used in rheumatoid arthritis (RA) in order to alleviate symptoms of joint inflammation, retard erosions and to treat extra-articular manifestations, although these drugs may increase the risk of bone mineral loss and osteoporotic fractures. To date, in Mexico there are no studies that identify the frequency of patients with RA with corticosteroids, receiving therapy for osteoporosis. Therefore, we evaluated the prevalence and factors related to the prescription of antiresorptives in 520 Mexican patients with RA. We used a multivariate model to identify variables associated with antiresorptives prescription. We identified that although 79% of patients were under treatment with glucocorticoids, only 13% received antiresorptive agents as preventive therapy for osteoporosis. The multivariate analysis identified that higher proportions of antiresorptive drugs prescriptions were associated with female patients (OR 11.40, 95% CI: 1.5-84.3, P = 0.02), an age of 40 years or more (OR 3.22, 95% CI: 1.3-8.3, P = 0.02) and to consume a lower number of cointerventions with other drugs (OR 1.09, 95% CI: 1.0-1.2, P = 0.03). Corticosteroid treatment was not associated with the prescription of antiresorptives (P = 0.31). In conclusion, a low proportion of Mexicans with RA receive antiresorptive therapy independently regardless of whether they consume or not chronically corticosteroids. Additional strategies should be evaluated to encourage the prevention and early treatment for osteoporosis in patients with RA.

Focused microwave-induced combustion for digestion of botanical samples and metals determination by ICP OES and ICP-MS.

The advantages and shortcomings of focused microwave-induced combustion (FMIC) for digestion of plant samples were studied. The effects of sample mass, absorbing solution, oxygen gas flow-rate, and time of reflux step on recoveries of major, minor and trace metals were systematically evaluated. Afterwards, Al, Ba, Ca, Co, Cr, Cu, Mg, Mn, Ni, Sr, V, and Zn were determined by inductively coupled plasma optical emission spectrometry (ICP OES) and by inductively coupled plasma mass spectrometry (ICP-MS). The main advantages of FMIC when compared to microwave-assisted wet digestion (MAWD) and focused-microwave-assisted wet digestion (FMAWD) are the possibility to digest larger masses of samples (up to 3g) using shorter heating times and diluted nitric acid solution for absorbing all analytes. Using the selected experimental conditions for FMIC, residual carbon content was lower than 0.7% for all samples and relative standard deviation (RSD) varied from 1.5 to 14.1%. Certified reference materials (NIST 1515 apple leaves and NIST 1547 peach leaves) were used for checking accuracy and determined values for all metals were in agreement with certified values at a 95% confidence level. No statistical difference (ANOVA, 95% of confidence level) was observed for results obtained by FMIC, FMAWD, and MAWD. Limits of detection were lower when using FMIC in the range of 0.02-0.15 µg g(-1) for ICP OES and 0.001-0.01 µg g(-1) for ICP-MS, which were about 3 and 6 times lower than the values obtained by FMAWD and MAWD, respectively. It is important to point out that FMIC was a suitable sample preparation method for major, minor and trace metals by both determination techniques (ICP OES and ICP-MS). Additionally, since it allows lower LODs (because up to 3g of sample can be digested) and diluted acid solutions are used (without any further dilution), the use of ICP-MS is not mandatory.

Cervical human papillomavirus infection in Mexican women with systemic lupus erythematosus or rheumatoid arthritis.

Cervical human papillomavirus (HPV+) infection is associated with an increased risk of cervical dysplasia. Although the frequency of HPV+ in systemic lupus erythematosus (SLE) has been investigated in some races its prevalence in Hispanic women is still unknown. This cross-sectional study evaluated the prevalence of cervical HPV+ in Mexican women with SLE (n = 34) or rheumatoid arthritis (RA) (n = 43) and in healthy controls (n = 146). These women were interviewed about risk factors for sexually transmitted infections and cervical cytology analysis was performed. HPV+ viral types were identified using PCR: HPV+ was observed in 14.7% of SLE, 27.9% of RA and 30.8% of controls. High-risk HPV types were observed in 11.7% of women with SLE, 27.9% of women with RA, and in 26% of the controls. High-risk viral types 58, 35 and 18 were the most frequently identified in SLE. Two women with SLE had a high-grade squamous intraepithelial lesion and one had cervical cancer. An association was observed between methotrexate utilization, longer duration of therapy with prednisone, and HPV+ in RA or SLE. Thus, there is a high prevalence of cervical HPV infection in Mexican women with SLE or RA, and physicians must be vigilant in preventing the development of cervical dysplasia.

Effect of wheat bran and flaxseed on cadmium effects and retention in rats.

Dietary fiber can affect cadmium (Cd) absorption and toxicity, but the effect appears to depend on the type of dietary fiber. The aim of the present study was to compare the effect of dietary sources containing distinct amounts of soluble and insoluble fiber on Cd absorption, accumulation and toxicity in growing rats. The absorption of essential macrominerals (Ca, P and Mg) was also evaluated. Animals received a nutritionally balanced diet with cellulose (cel - control), wheat bran or flaxseed as the fiber source with 0 or 50 mg Cd kg(-1) diet, during 30 days. Cd exposure reduced body weight gain, feed efficiency ratio, epididymal fat relative weight and liver relative weight, and increased plasma alanine aminotransferase activity in all fiber groups. The apparent Cd absorption was similar among Cd-groups, but the flax-Cd group had a higher hepatic and renal Cd concentration. Cd decreased the absorption of Ca and P, and increased Mg absorption in the wheat bran and flaxseed groups, but not in the cel group. Although the different fiber sources investigated had no effect on Cd toxicity, the major soluble fiber source, flaxseed, increased Cd retention. Thus, caution should be taken in the intake of flaxseed by Cd-exposed populations.

Delayed biochemical changes induced by mercury intoxication are prevented by zinc pre-exposure.

This work evaluated the delayed effects of mercury and the effectiveness of zinc in preventing such effects. Pups were pre-treated with 1 daily dose of ZnCl(2) (27 mg/kg/day, by subcutaneous injections) from 3rd to 7th postnatal day and received 1 daily dose of 5 mg/kg of HgCl(2), for 5 subsequent days (8-12 days old). Animals were euthanized 21 days after the end of Hg-exposure. Porphobilinogen-synthase activity as well as zinc and mercury contents was determined in the liver and kidneys. Alanine aminotransferase, aspartate aminotransferase and lactic dehydrogenase activities as well as urea, creatinine and glucose levels were analyzed in plasma or serum. Some animals were considered more sensitive to mercury, since they did not recover the body weight gain and presented an increase of renal and hepatic mercury content, urea and creatinine levels; a decrease in renal porphobilinogen-synthase and alanine aminotransferase activities, as well as a decrease in the liver and an increase in kidney weights. Some animals were considered less sensitive to mercury because they recovered the body weight and presented no biochemical alterations in spite of mercury in the tissues. Zinc prevents partially or totally the alterations caused by mercury even those that persisted for a long time after the end of exposure. These findings suggest that there is difference among the animals regarding the sensitivity to mercury.

Identification of rosmarinic acid as the major active constituent in Cordia americana.

ETHNOPHARMACOLOGICAL RELEVANCE: Preparation from leaves of Cordia americana have been widely used in traditional medicine in South Brazil to treat wounds and various inflammations. AIM OF THE STUDY: The objective of this work was to identify the effective compounds in the ethanolic extract prepared from the leaves of Cordia americana, which is used in traditional South Brazilian medicine as anti-inflammatory and wound healing remedy. MATERIALS AND METHODS: Isolation and structure elucidation techniques were performed in order to identify the compounds of Cordia americana and HPLC analysis was used for the quantification. The major constituent and the ethanolic extract were investigated for inhibition of 5-lipoxygenase, p38alpha MAPK, TNFalpha release and NF-kappaB as well as in the fibroblast scratch assay. RESULTS: Rosmarinic acid (1) was identified as the major compound with an amount of 8.44% in the ethanolic extract of the leaves of Cordia americana. The ethanolic extract as well as (1) exhibited the highest inhibitory effects on 5-lipoxygenase (IC(50)=0.69 and 0.97microg/mL, resp., IC50 of BWA4C as reference: 0.3microM) and p38alpha (IC50=3.25 and 1.16microg/mL, resp., IC50 of SB203580 as reference: 0.046microM) and moderate inhibitory effects on TNFalpha release. Slight effects were observed in the fibroblast scratch assay. CONCLUSIONS: This study increases our knowledge on the effective compound in Cordia americana and supports its use in traditional medicine. We demonstrated for the first time pharmacological effects of Cordia americana and we provide evidences for a crucial role of rosmarinic acid as the major key player.

Antioxidant system activation by mercury in Pfaffia glomerata plantlets.

Oxidative stress caused by mercury (Hg) was investigated in Pfaffia glomerata plantlets grown in nutrient solution using sand as substrate. Thirty-day-old acclimated plants were treated for 9 days with four Hg levels (0, 1, 25 and 50 microM) in the substrate. Parameters such as growth, tissue Hg concentration, toxicity indicators (delta-aminolevulinic acid dehidratase, delta-ALA-D, activity), oxidative damage markers (TBARS, lipid peroxidation, and H(2)O(2) concentration) and enzymatic (superoxide dismutase, SOD, catalase, CAT, and ascorbate peroxidase, APX) and non-enzymatic (non-protein thiols, NPSH, ascorbic acid, AsA, and proline concentration) antioxidants were investigated. Tissue Hg concentration increased with Hg levels. Root and shoot fresh weight and delta-ALA-D activity were significantly decreased at 50 microM Hg, and chlorophyll and carotenoid concentration were not affected. Shoot H(2)O(2) concentration increased curvilinearly with Hg levels, whereas lipid peroxidation increased at 25 and 50 microM Hg, respectively, in roots and shoots. SOD activity showed a straight correlation with H(2)O(2) concentration, whereas CAT activity increased only in shoots at 1 and 50 microM Hg. Shoot APX activity was either decreased at 1 microM Hg or increased at 50 lM Hg. Conversely, root APX activity was only increased at 1 microM Hg. In general, AsA, NPSH and proline concentrations increased upon addition of Hg, with the exception of proline in roots, which decreased. These changes in enzymatic and non-enzymatic antioxidants had a significant protective effect on P. glomerata plantlets under mild Hg-stressed conditions.

Influence of cereal bran supplement on cadmium effects in growing rats.

Strategies to diminish cadmium (Cd) absorption are highly desirable especially where Cd exposure due to environmental contamination is still inevitable. Cd toxicity may be influenced by dietary components, such as fiber and minerals. Multimixtures are low-cost cereal bran supplements used in Brazil and in other countries to counteract malnutrition in low-income populations. This study was aimed at evaluating whether multimixture would reduce Cd effects in young rats. Animals received a diet with or without the multimixture plus 0, 5, or 25 mg Cd/kg (control, Cd-5, and Cd-25 groups) during 30 days. The Cd-5 groups were similar to control groups in all parameters analyzed, except in the higher renal Cd concentration. However, the Cd-25 groups had lower biological growth parameters and renal delta-aminolevulinate dehydratase activity, besides higher renal Cd concentration and plasma alanine aminotransferase activity compared to the controls. The multimixture did not prevent Cd effects in the Cd-25 group, but caused a small reduction in renal Cd concentration in the Cd-5 group. Although this multimixture was ineffective to prevent Cd effects at the higher concentration, it seemed to reduce Cd accumulation at the lower Cd dietary concentration, which is similar to levels of human exposure in some polluted areas.