Kazimierz Wrobel

Capillary electrophoresis-inductively coupled plasma-mass spectrometry: an attractive complementary technique for elemental speciation analysis.

Some basic and practical aspects of interfacing capillary electrophoresis to inductively coupled plasma-mass spectrometry (CE-ICP-MS) are reviewed in this article with emphasis on the use of this hyphenated technique for elemental speciation analysis. The principles behind the techniques of both CE and ICP-MS are introduced. The interfacing of CE to ICP-MS is discussed including several devices and nebulizers reported in literature. A brief account of their advantages and limitations is given. The various CE-ICP-MS applications for elemental speciation analysis are also reviewed. Some issues concerning the future of CE-ICP-MS for the elemental speciation analyses are discussed.

Determination of As(III), As(V), monomethylarsonic acid, dimethylarsinic acid and arsenobetaine by HPLC-ICP-MS: analysis of reference materials, fish tissues and urine

In this study, a rapid and sensitive high performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) determination of primary As species in fish tissues and urine is reported. The separation was achieved on an Altima C18 column with a mobile phase containing citric acid and hexanesulfonic acid (pH 4.5). As(V), monomethylarsonic acid (MMA), As(III), dimethylarsinic acid (DMA) and arsenobetaine (AsB) were separated in less than 4 min with retention times of 83, 99, 130, 166 and 208 s, respectively. This separation of five species in less than 4 min should be attractive to those interested in As speciation. The quantification limits were 44, 56, 94, 64, 66 ng l(-1) and the relative standard deviations (R.S.D.) for day-to-day injections of As at 2 mug l(-1) were 2.0, 3.1, 2.4, 3.8 and 4.0%. The procedure was tested using two reference materials (DORM-2 dogfish muscle tissue, NIST SRM 2670 Freeze-dried Urine, normal level) and then applied to real-world samples. The results obtained demonstrate the suitability of the procedure for screening and quantification at physiological levels of primary As species in biological samples.

Dietary Advanced Glycation End Products and Their Role in Health and Disease

Over the past 2 decades there has been increasing evidence supporting an important contribution from food-derived advanced glycation end products (AGEs) to the body pool of AGEs and therefore increased oxidative stress and inflammation, processes that play a major role in the causation of chronic diseases. A 3-d symposium (1st Latin American Symposium of AGEs) to discuss this subject took place in Guanajuato, Mexico, on 1-3 October 2014 with the participation of researchers from several countries. This review is a summary of the different presentations and subjects discussed, and it is divided into 4 sections. The first section deals with current general knowledge about AGEs. The second section dwells on mechanisms of action of AGEs, with special emphasis on the receptor for advanced glycation end products and the potential role of AGEs in neurodegenerative diseases. The third section discusses different approaches to decrease the AGE burden. The last section discusses current methodologic problems with measurement of AGEs in different samples. The subject under discussion is complex and extensive and cannot be completely covered in a short review. Therefore, some areas of interest have been left out because of space. However, we hope this review illustrates currently known facts about dietary AGEs as well as pointing out areas that require further research.

Determination of total aluminum, chromium, copper, iron, manganese, and nickel and their fractions leached to the infusions of black tea, green tea, Hibiscus sabdariffa, and Ilex paraguariensis (mate) by ETA-AAS.

Total aluminum, chromium, copper, iron, manganese, and nickel were determined in black tea, green tea, Hibiscus sabdariffa, and Ilex paraguariensis (mate) by electrothermal atomic absorption spectrometry after nitric/perchloric acid digestion. In each case, one ground sample of commercially available leafy material was prepared and three 0.5-g subsamples were run in parallel. The infusions were also analyzed and the percentage of each element leached into the liquor was evaluated. The obtained results indicated that hibiscus and mate contained lower levels of aluminum (272±19 µg/g and 369±22 µg/g, respectively) as referred to black tea (759±31 µg/g) or green tea (919±29 µg/g) and suggested that mate drinking could be a good dietary source of essential micronutrient manganese (total content 2223±110 µg/g, 48.1% leached to the infusion). It was also found that the infusion of hibiscus could supply greater amounts of iron (111±5 µg/g total, 40.5% leached) and copper (5.9±0.3 µg/g total, 93.4% leached) as compared to other infusions. Moreover, it was found that the percentage of element leached to the infusion was strongly related to the tannins content in the beverage (correlation coefficients >0.82 with the exception for nickel); for lower tannins level, better leaching was observed.

Aluminium and silicon speciation in human serum by ion-exchange high-performance liquid chromatography–electrothermal atomic absorption spectrometry and gel electrophoresis

Speciation of aluminium and silicon in serum was studied by a reliable and sensitive high-performance liquid chromatographic-electrothermal atomic absorption spectrometric (HPLC-ETAAS) hybrid method, based on the use of a polymeric anion-exchange column (Protein-Pak DEAE-5PW). This polymer-based column minimizes the risk of aluminium losses and of silicon contamination from the column during separation. The results obtained were compared with the results of previous studies carried out using different, complementary techniques including ultramicrofiltration, gel filtration and silica-based column for HPLC. In order to ascertain which protein(s) of serum actually bind(s) aluminium, gel electrophoresis was employed for the further separation of the column fractions obtained by HPLC and aluminium was determined in separate aliquots of the same fractions. From all the experiments, it appears that transferrin (Tf) is the only serum protein that binds aluminium and it contains about 90% of total serum aluminium. It was also confirmed that in the presence of desferrioxamine (DFO). aluminium is partly displaced from its complex with transferrin to a low molecular mass AL-DFO complex. Aluminum citrate seems to be the main low molecular mass aluminium species in serum, amounting to about (12 +/- 5% of the total aluminium in an aluminium-loaded serum sample. The proposed speciation procedure permits the simultaneous identification and determination of three aluminium species in metal-spiked serum (Al-Tf, Al-DFO and AI-citrate). The result for silicon suggest that it seems to be unspecifically adsorbed to several serum proteins and its speciation is not affected by the presence of DFO.(ABSTRACT TRUNCATED AT 250 WORDS)

Trace elements status in diabetes mellitus type 2: Possible role of the interaction between molybdenum and copper in the progress of typical complications

It is well established that both, the deficiency and possible overload of mineral micronutrients have adverse health effects. It is also generally accepted that non-essential xenobiotics contribute to oxidative damage, which is considered one of the principal factors in diabetes and its complications. The purpose of this work was to gain an insight on the global role of metal/metalloids in the progress of diabetes mellitus type 2. In such approach, aluminum, vanadium, chromium, manganese, cobalt, nickel, copper, zinc, arsenic, selenium, molybdenum, mercury, cadmium and lead were determined by inductively coupled plasma-mass spectrometry (ICP-MS) in serum and urine of 76 diabetic patients (age 52 ± 8 years, 5-16 years of DM2, 52 subjects with slight-to-moderate complications and 24 with severe complications). A series of anthropometric and clinical parameters usually evaluated in the follow-up of patients were assessed by standard methods. Statistical analysis (unpaired t-test, analysis of correlation and principal component analysis) was then carried out in search of possible relationships existing among metals/metalloids and these parameters. The results obtained suggest that antagonistic interaction between molybdenum and copper might be involved in the progress of diabetes complications.

Serum selenium and glutathione peroxidase concentrations in type 2 diabetes mellitus patients

AIMS Antioxidant selenium (Se) properties and, its protective role against oxidative damage play an important role in diabetic complications. Our objective was to gain further insight on a link between selenium status and diabetic nephropathy. METHODS We assessed glutathione peroxidase (GPx) and Se in type 2 diabetes mellitus patients with microalbuminuria (MA) (group 1), without microalbuminuria (group 2), and in control subjects (group 3). Glucose, urea, creatinine and glycated hemoglobin tests were tested in sera. A complete clinical record was elaborated. RESULTS For diabetic patients both, the time from diagnosis and plasma glucose concentration were higher in group 1 as compared to group 2. Control group showed higher serum Se concentrations as compared to the diabetic groups. The two groups of diabetic patients showed similar serum Se levels. Serum concentration of GPx was significantly lower in group 1 as compared to groups 2 and 3. Microalbuminuria (MA) test showed a positive correlation with glucose, and a negative relationship with serum Se and GPx. Multiple regression revealed an inverse relationship between selenium or GPx in serum and the results of the MA test. CONCLUSIONS Our results suggest that lower Se and GPx levels in diabetic patients may be implicated in the diabetic nephropathy.

Methylome analysis reveals an important role for epigenetic changes in the regulation of the Arabidopsis response to phosphate starvation

Significance Significant progress has been achieved in our understanding of plant adaptive responses to ensure growth and reproduction in soils with low phosphate (Pi) availability. However, the potential role of epigenetic mechanisms in the modulation of these responses remains largely unknown. In this article, we describe dynamic changes in global DNA methylation patterns that occur in Arabidopsis plants exposed to low Pi availability; these changes are associated with the onset of Pi starvation responses. We show that the expression of a subset of low Pi-responsive genes is modulated by methylation changes and that DNA methylation is required for the proper establishment of developmental and molecular responses to Pi starvation. Phosphate (Pi) availability is a significant limiting factor for plant growth and productivity in both natural and agricultural systems. To cope with such limiting conditions, plants have evolved a myriad of developmental and biochemical strategies to enhance the efficiency of Pi acquisition and assimilation to avoid nutrient starvation. In the past decade, these responses have been studied in detail at the level of gene expression; however, the possible epigenetic components modulating plant Pi starvation responses have not been thoroughly investigated. Here, we report that an extensive remodeling of global DNA methylation occurs in Arabidopsis plants exposed to low Pi availability, and in many instances, this effect is related to changes in gene expression. Modifications in methylation patterns within genic regions were often associated with transcriptional activation or repression, revealing the important role of dynamic methylation changes in modulating the expression of genes in response to Pi starvation. Moreover, Arabidopsis mutants affected in DNA methylation showed that changes in DNA methylation patterns are required for the accurate regulation of a number of Pi-starvation–responsive genes and that DNA methylation is necessary to establish proper morphological and physiological phosphate starvation responses.

New insights into somatic embryogenesis: leafy cotyledon1, baby boom1 and WUSCHEL-related homeobox4 are epigenetically regulated in Coffea canephora.

Plant cells have the capacity to generate a new plant without egg fertilization by a process known as somatic embryogenesis (SE), in which differentiated somatic cells can form somatic embryos able to generate a functional plant. Although there have been advances in understanding the genetic basis of SE, the epigenetic mechanism that regulates this process is still unknown. Here, we show that the embryogenic development of Coffea canephora proceeds through a crosstalk between DNA methylation and histone modifications during the earliest embryogenic stages of SE. We found that low levels of DNA methylation, histone H3 lysine 9 dimethylation (H3K9me2) and H3K27me3 change according to embryo development. Moreover, the expression of LEAFY COTYLEDON1 (LEC1) and BABY BOOM1 (BBM1) are only observed after SE induction, whereas WUSCHEL-RELATED HOMEOBOX4 (WOX4) decreases its expression during embryo maturation. Using a pharmacological approach, it was found that 5-Azacytidine strongly inhibits the embryogenic response by decreasing both DNA methylation and gene expression of LEC1 and BBM1. Therefore, in order to know whether these genes were epigenetically regulated, we used Chromatin Immunoprecipitation (ChIP) assays. It was found that WOX4 is regulated by the repressive mark H3K9me2, while LEC1 and BBM1 are epigenetically regulated by H3K27me3. We conclude that epigenetic regulation plays an important role during somatic embryogenic development, and a molecular mechanism for SE is proposed.

Selenium speciation in low molecular weight fraction of Se-enriched yeasts by HPLC-ICP-MS: detection of selenoadenosylmethionine

In this study, selenium speciation in low molecular weight (LMW) fraction of Se-enriched yeasts was investigated assuming the presence of selenoadenosylmethionine (AdoSeMet). In order to minimize the risk of species decomposition, the pretreatment procedure consisted of homogenization of fresh cells and precipitation of proteins with perchloric acid at 0 °C. The species separation was achieved by ion pairing HPLC (hexanesulfonic acid) with UV (254 nm) and ICP-MS detection. The assignment of chromatographic peaks was by spiking the sample with both the commercial and synthesized standards. The synthesis of AdoSeMet and selenoadenosylhomocysteine (AdoSeHcy) was performed and their structures confirmed by ES-MS analysis. These two selenium species were identified in yeasts extract and the presence of Se-cystine was also observed. Significantly lower ratio between AdoSeMet and AdoSeHcy was observed in yeast extracts as compared to the ratio between sulfur analogs. This observation could contribute to further understanding of the selenium role in biomethylation processes.