Quantification of modified nucleotides and nucleosides by isotope dilution mass spectrometry.

Epigenetic modifications are closely related to certain disorders of the organism, including the development of tumors. One of the main epigenetic modifications is the methylation of DNA cytosines, 5-methyl-2'-deoxycycytidine. Furthermore, 5-mdC can be oxidized to form three new modifications, 5-(hydroxymethyl)-2'-deoxycytidine, 5-formyl-2'-deoxycytidine, and 5-carboxy-2'-deoxycytidine. The coupling of liquid chromatography with tandem mass spectrometry has been widely used for the total determination of methylated DNA cytosines in samples of biological and clinical interest. These methods are based on the measurement of the free compounds (e.g., urine) or after complete hydrolysis of the DNA (e.g., tissues) followed by a preconcentration, derivatization, and/or clean-up step. This review highlights the main advances in the quantification of modified nucleotides and nucleosides by isotope dilution using isotopically labeled analogs combined with liquid or gas chromatography coupled to mass spectrometry reported in the last 20 years. The different possible sources of labeled compounds are indicated. Special emphasis has been placed on the different types of chromatography commonly used (reverse phase and hydrophilic interaction liquid chromatography) and the derivatization methods developed to enhance chromatographic resolution and ionization efficiency. We have also revised the application of bidimensional chromatography and indicated significant biological and clinical applications of these determinations.

Evaluation of different isotope dilution mass spectrometry strategies for the characterization of naturally abundant and isotopically labelled peptide standards.

Natural abundance and isotopically labelled tryptic peptides are routinely employed as standards in quantitative proteomics. The certification of the peptide content is usually carried out by amino acid analysis using isotope dilution mass spectrometry (IDMS) after the acid hydrolysis of the peptide. For the validation and traceability of the amino acid analysis procedure, expensive certified peptides must be employed. In this work we evaluate different IDMS alternatives which will reduce the amount of certified peptide required for validation of the amino acid analysis procedure. In this context, the characterization of both natural and isotopically labelled synthetic angiotensin I peptides was carried out. First, we applied a fast procedure for peptide hydrolysis based on microwave-assisted digestion and employed two certified peptide reference materials SRM 998 angiotensin I and CRM 6901-b C-peptide for validation of the hydrolysis procedure. The amino acids proline, leucine, isoleucine, valine, tyrosine, arginine and phenylalanine were evaluated for their suitability for peptide certification by IDMS by both liquid chromatography with tandem mass spectrometry (LC-MS/MS) and gas chromatography with mass spectrometry (GC)-MS/MS. Then, natural angiotensin I and 13C1-labelled angiotensin I were synthesized in-house and purified by preparative liquid chromatography. The concentration of the 13C1-labelled angiotensin I peptide was established by reverse IDMS in its native form using SRM 998 angiotensin I as reference. The concentration of the natural synthesized peptide was determined by IDMS both using the 13C1-labelled peptide in its native form and by amino acid analysis showing comparable results. Finally, the synthetic naturally abundant angiotensin I peptide was employed as "in-house" standard for the validation of subsequent peptide characterization procedures. Therefore, the novelty of this work relies on, first, the development of a faster hydrolysis procedure assisted by focused microwaves, providing complete hydrolysis in 150 min, and secondly, a validation strategy combining GC-MS and LC-MS/MS that allowed us to certify the purity of an in-house-synthesized peptide standard that can be employed as quality control in further experiments.

Determination of 3-monoiodotyrosine and 3,5-diiodotyrosine in newborn urine and dried urine spots by isotope dilution tandem mass spectrometry.

High levels of 3-mono- and 3,5-diiodotyrosine (MIT and DIT, respectively) in urine have been related to iodotyrosine dehalogenase 1 deficiency, a type of congenital hypothyroidism. However, the determination of MIT and DIT in urine is not included in newborn screening programs performed in clinical laboratories to detect inborn errors of metabolism. We report here on the development of an analytical method for the determination of MIT and DIT in newborn urine and dried urine spots (DUS) by Liquid Chromatography Isotope Dilution tandem Mass Spectrometry (LC-IDMSMS). The development included the synthesis of 15N-monoiodotyrosine and 13C2-diiodotyrosine through the iodination of 15N-tyrosine and 13C2-tyrosine, respectively, using bis(pyridine)iodonium(I) tetrafluoroborate (IPy2BF4). Both labelled analogues were added at the beginning of the sample preparation procedure and used to develop both single- and double-spike LC-IDMS methods for the determination of MIT and DIT. The developed double spike methodology was able to quantify and correct possible MIT ↔ DIT interconversions throughout the sample preparation, which was observed for concentrated urine samples but not for DUS. Suppression matrix effects on the absolute signals of MIT and DIT were observed in urine samples but did not affect the IDMS results as recoveries on urine samples at different dilution factors could be considered quantitative. Method detection limits were 0.018 and 0.046 ng g-1 (limits of quantification 0.06 and 0.15 ng g-1) by single-spike IDMS, for MIT and DIT, respectively, in the analysis of urine samples and 0.07 and 0.05 ng g-1 (limits of quantification 0.23 and 0.17 ng g-1) for MIT and DIT, respectively, in the analysis of DUS. No significant differences were obtained for MIT concentrations in the analysis of the same newborn samples stored as liquid urine or DUS when the results were corrected for the creatinine content. Finally, 36 DUS samples from healthy newborns were analyzed and MIT was detected in all samples at low ng mg-1creatinine levels.

The Elderly Nursing Core Set and the cognition of Portuguese older adults: a cross-sectional study.

BACKGROUND: The aging population and its associated health needs require specific nursing care. The aim of this study was to draw an epidemiological profile of Portuguese elderly adults attending in residential homes and day centers and to evaluate the association between the functioning and cognition of these older adults and their sociodemographic characteristics and presence of multimorbidity. METHODS: This was a cross-sectional study of 613 older adults. Functioning was assessed using the Elderly Nursing Core Set, and cognition was assessed using the Mini Mental State Examination. Descriptive and inferential analyses were performed. RESULTS: The mean age was 85.73 years; the majority of the participants were female (69.3 %), widowed (67.0 %) and over 85 years old (60.4 %). A total of 68.2 % of the sample presented multimorbidity. A total of 54.5 % had cognitive impairment, and the average functional profile was classified as "moderate difficulty". Institutionalized older adults had more diseases than those who attended the day center. Women, those who were illiterate, those who were institutionalized and older adults who had diseases of the nervous system had a worse functional profile and greater cognitive impairment. Those with multimorbidity had a worse functional profile, and those without a spouse had greater cognitive impairment. CONCLUSIONS: Given the functional and cognitive profile of older adults, it is necessary to adopt care practices focused on the rehabilitation/maintenance of self-care and affective relationships. This care must be provided by highly qualified professionals. Therefore, it is necessary to increase the ratio of nurses per older adult in these institutions.

Quantitative Assessment of Individual Populations Present in Nanoparticle-Antibody Conjugate Mixtures Using AF4-ICP-MS/MS.

A current remaining challenge in nanotechnology is the fast and reliable determination of the ratios between engineered nanoparticles and the species attached to their surface after chemical functionalization. The approach proposed herein based on the online coupling of asymmetric flow field-flow fractionation (AF4) with inductively coupled plasma-tandem mass spectrometry (ICP-MS/MS) allows for the first time the direct determination of such ratios in CdSe/ZnS core-shell quantum dot:rat monoclonal IgG2a antibody (QD:Ab) conjugate mixtures in a single run without any previous sample preparation (i.e., derivatization). AF4 provides full recovery and adequate resolution of the resulting bioconjugate from the excess of nanoparticles and proteins used in the different bioconjugation mixtures (1:1, 2:1, and 3:1 QD:Ab molar ratios were assessed). The point-by-point determination by ICP-MS/MS of the metal to sulfur ratios along the bioconjugate fractographic peak allowed disclosing the mixture of the different species in the bioconjugated sample, providing not only the limits of the range of QD:Ab ratios in the different bioconjugate species resulting after functionalization but also a good estimation of their individual relative abundance in the mixture. Interestingly, a wide variety of compositions were observed for the different bioconjugate mixtures studied (QD:Ab molar ratios ranging from 0.27 to 4.6). The resulting weighted QD:Ab ratio computed in this way for each bioconjugate peak matches well with both the global (average) QD:Ab ratio experimentally obtained by the simpler peak area ratio computation and the theoretical QD:Ab molar ratios assayed, which internally validates the procedure developed.

Anion-Specific Sulfur Isotope Analysis by Liquid Chromatography Coupled to Multicollector ICPMS.

An accurate method has been developed to measure, in a single analytical run, δ34S in sulfite, sulfate and thiosulfate in water samples by liquid chromatography combined with multicollector inductively coupled plasma mass spectrometry (MC-ICPMS). The method is based on the anionic exchange separation of sulfur species prior to their online isotope ratio determination by MC-ICPMS. Mass bias correction was accomplished by a novel approach based on the addition of an internal sulfur-containing standard to the sample. This innovative approach was compared to the sample-standard bracketing procedure. On-column isotopic fractionation was observed and therefore corrected by external calibration. Isotopic ratios were calculated by linear regression slope (LRS), an advantageous method for transient signals, leading to a combined uncertainty of δ34S below 0.25‰ and a reproducibility below 0.5‰ for the injection of 1 µg of S. The method was successfully applied to the measurement of δ34S in synthetic solutions and environmental water samples. Matrix effects leading to δ34S overestimation were observed for sulfate in some samples with high sodium/sulfate mass ratios. The developed analytical procedure simplifies the δ34S analysis of liquid environmental samples since preparation steps are no longer required and allows the analysis of several sulfur-containing species in a single run.

Isotopically Enriched Tracers and Inductively Coupled Plasma Mass Spectrometry Methodologies to Study Zinc Supplementation in Single-Cells of Retinal Pigment Epithelium in Vitro.

Mass spectrometry-based techniques, such as inductively coupled plasma mass spectrometry (ICPMS) and laser ablation (LA) ICPMS, combined with an isotope pattern deconvolution mathematical tool are proposed for a better understanding of supplementation studies in cultured cells. An in vitro model of human retinal pigment epithelium (HRPEsv) cells was treated with different concentrations (0-150 µm Zn, 1 mL) of enriched stable isotope tracers of Zn in the form of sulfate and/or gluconate. Supplementations with t68ZnSO4 or t70Zn-gluconate alone and in combination (1:1 molar ratio) were investigated to evaluate the exogenous contribution and distribution of Zn in the treated cells. In order to obtain not only the Zn concentration for a cell population (mineralized cells) but also single cell information about the contribution of exogenous Zn and their distribution within micrometer cells structures, LA-ICPMS was employed to directly analyze cryopreserved cells. natZn, t68Zn, and t70Zn molar fraction images obtained from cells and cell aggregates allowed confirming the uptake of exogenous Zn by HRPEsv cells, being t68Zn and t70Zn molar fractions close to 1 in the cell nuclei. Under the selected experimental conditions tested (24 h treatments), no significant differences were obtained in the Zn distribution depending on its chemical form.

Systematic Literature Review of Food-Intake Monitoring in an Aging Population.

The dietary habits of people directly impact their health conditions. Especially in elder populations (in 2017, 6.7% of the world's population was over 65 years of age), these habits could lead to important-nutrient losses that could seriously affect their cognitive and functional state. Recently, a great research effort has been devoted to using different technologies and proposing different techniques for monitoring food-intake. Nevertheless, these techniques are usually generic but make use of the most innovative technologies and methodologies to obtain the best possible monitoring results. However, a large percentage of elderly people live in depopulated rural areas (in Spain, 28.1% of the elderly population lives in this type of area) with a fragile cultural and socioeconomic context. The use of these techniques in these environments is crucial to improving this group's quality of life (and even reducing their healthcare expenses). At the same time, it is especially challenging since they have very specific and strict requirements regarding the use and application of technology. In this Systematic Literature Review (SLR), we analyze the most important proposed technologies and techniques in order to identify whether they can be applied in this context and if they can be used to improve the quality of life of this fragile collective. In this SLR, we have analyzed 326 papers. From those, 29 proposals have been completely analyzed, taking into account the characteristics and requirements of this population.

A Novel Routing Scheme for Creating Opportunistic Context-Virtual Networks in IoT Scenarios.

The massive amount of traffic required by the emerging Internet of Things (IoT) paradigm can be supported by the imminent arrival of 5G next-generation networks. However, the limited capacity of resources in IoT nodes, e.g., battery lifetime or buffer space, opens a challenge to be taken into account when proposing new routing solutions on IoT scenarios with intermittent connectivity. In this paper, we propose the concept of Opportunistic Context-Virtual Networks (OCVNs). The novelty of this approach is to create virtual groups of nodes that share interests in common for routing purposes. Therefore, only the nodes that are interested in the content of the messages that are flowing throughout the network are used as relaying nodes, providing their own resources for the sake of the communication. By leveraging the use of store-carry-and-forward mechanisms, a novel routing algorithm is proposed and evaluated over two realistic scenarios. Experimental results reveal that our solution outperforms other well-known opportunistic routing algorithms in terms of delivery probability and overhead ratio, while resource usage of relaying nodes is significantly reduced.

Development of a Common Procedure for the Determination of Methylmercury, Ethylmercury, and Inorganic Mercury in Human Whole Blood, Hair, and Urine by Triple Spike Species-Specific Isotope Dilution Mass Spectrometry.

We report the first common methodology for the simultaneous determination of methylmercury (MeHg), ethylmercury (EtHg), and inorganic mercury (Hg(II)) in human blood hair and urine. With the exception of the initial sample mass (0.15 g for blood, 0.5 g for urine, and 0.1 g for hair), the same sample preparation and gas chromatography-inductively coupled plasma mass spectrometry (GC-ICPMS) measurement conditions are employed for the three matrixes providing experimental values in agreement with the certified values in the analysis of NIST SRM 955c (Caprine Blood) Level 3 and the certified human hairs IAEA 085 and IAEA 086. Also, the method provides quantitative recoveries for the three Hg species in the analysis of fortified human urine samples at 1, 2, and 5 ng Hg g-1. Mercury species concentrations for levels 2 and 4 of SRM 955c are reported here for the first time. A systematic interconversion of EtHg into Hg(II) was obtained for all matrixes reaching values up to 95% in blood, 29% in hair, and 11% in urine. MeHg dealkylation was also observed in a lesser extent in blood and hair analyses, but it was not observed when analyzing urine samples. Hg methylation was not observed in any matrix. The amount of NaBPr4 added for derivatization has been found to be the main factor responsible for Hg species interconversion. This work demonstrates for the first time that experimental conditions optimized for SRM 955c (caprine blood) are not valid for human blood samples as the optimum initial sample amount for a real sample is more than 3 times lower than that for SRM 955c.

Melatonin Decreases Glucose Metabolism in Prostate Cancer Cells: A 13C Stable Isotope-Resolved Metabolomic Study.

The pineal neuroindole melatonin exerts an exceptional variety of systemic functions. Some of them are exerted through its specific membrane receptors type 1 and type 2 (MT1 and MT2) while others are mediated by receptor-independent mechanisms. A potential transport of melatonin through facilitative glucose transporters (GLUT/SLC2A) was proposed in prostate cancer cells. The prostate cells have a particular metabolism that changes during tumor progression. During the first steps of carcinogenesis, oxidative phosphorylation is reactivated while the switch to the "Warburg effect" only occurs in advanced tumors and in the metastatic stage. Here, we investigated whether melatonin might change prostate cancer cell metabolism. To do so, 13C stable isotope-resolved metabolomics in androgen sensitive LNCaP and insensitive PC-3 prostate cancer cells were employed. In addition to metabolite 13C-labeling, ATP/AMP levels, and lactate dehydrogenase or pentose phosphate pathway activity were measured. Melatonin reduces lactate labeling in androgen-sensitive cells and it also lowers 13C-labeling of tricarboxylic acid cycle metabolites and ATP production. In addition, melatonin reduces lactate 13C-labeling in androgen insensitive prostate cancer cells. Results demonstrated that melatonin limits glycolysis as well as the tricarboxylic acid cycle and pentose phosphate pathway in prostate cancer cells, suggesting that the reduction of glucose uptake is a major target of the indole in this tumor type.

Long-term care units: a Portuguese study about the functional profile.

Aim: In this study, we analyze the relationship between the functional profile of older people admitted to long-term care units in Portugal and some demographic variables such as education level, sex, and age as well as the emotional state of mind. Methods: A sample of 59,516 older people from the National Network of Integrated Continuous Care of Portugal were analyzed in this longitudinal study. All the retrospective data of the older people were collected during the period of hospitalization at the long-term care units. The database records of these units were analyzed, and a functional profile spanning the period of hospitalization was calculated. Results: Activities of daily living and cognitive states improved, in the first 90 days of hospitalization, while mobility and instrumental activities of daily living worsened for the same period of 90 days. Generally, there was a decline in all domains after 450 days of hospitalization. The older women that did not attend school, those over 85 years old, and those who suffered from anxiety were pre-dominantly placed in the group of those with greater dependence (severe/complete dependence). Conclusion: The participants hospitalized between 90 and 360 days presented the best results in the long-term care units of the National Network of Integrated Continuous Care of Portugal. With this study, we highlight the importance of evaluating the functional status of persons in long-term hospitalizations and the influence exerted by the level of education on the recovery and rehabilitation of dependence.

Lessons Learned: Occupational Therapy in Nursing Homes during the First Wave of COVID-19 in Spain.

This research aims to explore the perception that occupational therapists working in elderly care facilities have about the measures implemented against the COVID-19 pandemic in their resources, and the impact that these measures have had on occupational therapy in these facilities. An interpretive paradigm was selected, using a qualitative approach and a phenomenological design. Sixteen occupational therapists working in elderly care facilities in two Spanish regions were included. Data were collected through semi-structured interviews. A discourse analysis of the narrative information was carried out using open, axial, and selective coding processes and the constant comparison method. Four themes were extracted from the analysis results: The initial chaos in senior centers; The blurring of occupational therapists' roles; The emergence of technology; and organizational and therapeutic proposals for future pandemics. The pandemic had a significant impact on the care and therapeutic processes in elderly care facilities. Occupational therapists had to stop performing their functions to dedicate themselves to other support, auxiliary or communication tasks between the center and the families. Similarly, it is worth noting the emergence of low-cost technology to facilitate communication and to carry out some therapeutic interventions.

Androgen-Dependent Prostate Cancer Cells Reprogram Their Metabolic Signature upon GLUT1 Upregulation by Manganese Superoxide Dismutase.

Prostate cancer is the second leading cause of cancer in men across the globe. The prostate gland accounts for some unique glycolytic metabolic characteristics, which causes the metabolic features of prostate tumor initiation and progression to remain poorly characterized. The mitochondrial superoxide dismutase (SOD2) is one of the major redox metabolism regulators. This study points out SOD2 as one major regulator for both redox and glycolytic metabolism in prostate cancer. SOD2 overexpression increases glucose transporter GLUT-1 and glucose uptake. This is not an insulin-mediated effect and seems to be sex-dependent, being present in male mice only. This event concurs with a series of substantial metabolic rearrangements at cytoplasmic and mitochondrial level. A concomitant decrease in glycolytic and pentose phosphate activity, and an increase in electron transfer in the mitochondrial electronic chain, were observed. The Krebs Cycle is altered to produce amino-acid intermediates by decreasing succinate dehydrogenase. This in turn generates a 13-fold increase in the oncometabolite succinate. The protein energy sensor AMPK is decreased at basal and phosphorylated levels in response to glucose deprivation. Finally, preliminary results in prostate cancer patients indicate that glandular areas presenting high levels of SOD2 show a very strong correlation with GLUT-1 protein levels (R2 = 0.287 p-value < 0.0001), indicating that in patients there may exist an analogous phenomenon to those observed in cell culture and mice.

Potential of Nassarius nitidus for monitoring organotin pollution in the lagoon of Bizerta (northern Thunisia).

Imposex and butyltin burden were assessed in Nassarius nitidus, Bolinus brandaris and Hexaplex trunculus collected at five stations in the Bizerta lagoon. Biological analysis showed that imposex followed type (a) in N. nitidus (distal evolution), against type (d) in the two muricids (proximal evolution). Imposex indices were higher in sites located nearby sources of tributyltin and N. nitidus was the least affected species of the five sites, followed by B. brandaris and H. trunculus. Butyltin analysis showed lower accumulation in N. nitidus followed by H. trunculus and B. brandaris. This study has allowed the gathering of data on imposex in a snail studied for the first time in Tunisia (N. nitidus). It suggests the possibility of using such snail as a complementary species for organotin monitoring programs in the Mediterranean and further confirmed that H. trunculus is the most suitable species for such investigations.

Functioning and Cognition of Portuguese Older Adults Attending in Residential Homes and Day Centers: A Comparative Study.

The functioning and cognition of older adults can be influenced by different care contexts. We aimed to compare the functioning profiles and cognition of institutionalized and noninstitutionalized older adults and to evaluate the effect of sociodemographic factors on the functioning and cognition. This is a cross-sectional study that included 593 older adults. The data were collected using the Elderly Nursing Core Set and Mini Mental State Examination. Women, older adults who did not attend school and those live in Residential Homes are more likely to have a higher degree of cognitive impairment than men, those who attended school and those frequent Day Centre. The chances of an older adult with moderate or severe cognitive impairment increases with age. Older women, older adults who did not attend school, and older adults who live in Residential Homes had a higher degree of functional problem than men, those who attended school and those who frequent a Day Centre, independently to age. It is necessary to promote the health literacy of older adults throughout life. The implementation of social and health responses should allow older adults to remain in their homes, given the influence of functioning and cognition on self-care and quality of life.

Treatment Adherence in Chronic Conditions during Ageing: Uses, Functionalities, and Cultural Adaptation of the Assistant on Care and Health Offline (ACHO) in Rural Areas.

The increasingly common scenario of an ageing population is related to a rise in the prevalence of problems associated with chronic conditions and comorbidities. Polypharmacy is frequent among this population, and it is a situation that can create medication management and adherence issues. This article introduces the features and functionalities of a voice assistant (Assistant on Health and Care Offline, ACHO) that aims to facilitate treatment adherence among elderly adults. Specifically adapted for its use in rural contexts, it does not require an Internet connection. Its development consisted of two stages: a first stage of problem diagnosis, in which the classic tools of ethnographic fieldwork were used, and a second stage of design implementing methodologies developed by Ambient Assisted Living (AAL) programmes. The main design characteristic of this new digital care system is that it is adapted to the needs of its end-users. It includes features such as voice customisation and the personal identification of medication, it can be connected to other digital devices, and information is introduced and supervised by healthcare professionals. These custom features introduce a safer medication administration procedure, improve supervision strategies, and increase patients' trust in the prescription process.

Identification of a tri-iron(III), tri-citrate complex in the xylem sap of iron-deficient tomato resupplied with iron: new insights into plant iron long-distance transport.

The identification of Fe transport forms in plant xylem sap is crucial to the understanding of long-distance Fe transport processes in plants. Previous studies have proposed that Fe may be transported as an Fe-citrate complex in plant xylem sap, but such a complex has never been detected. In this study we report the first direct and unequivocal identification of a natural Fe complex in plant xylem sap. A tri-Fe(III), tri-citrate complex (Fe(3)Cit(3)) was found in the xylem sap of Fe-deficient tomato (Solanum lycopersicum Mill. cv. 'Tres Cantos') resupplied with Fe, by using an integrated mass spectrometry approach based on exact molecular mass, isotopic signature and Fe determination and retention time. This complex has been modeled as having an oxo-bridged tri-Fe core. A second complex, a di-Fe(III), di-citrate complex was also detected in Fe-citrate standards along with Fe(3)Cit(3), with the allocation of Fe between the two complexes depending on the Fe to citrate ratio. These results provide evidence for Fe-citrate complex xylem transport in plants. The consequences for the role of Fe to citrate ratio in long-distance transport of Fe in xylem are also discussed.

The Quality of Life of Older Individuals Following the World Health Organization Assessment Criteria.

The aim of this study was to evaluate the psychometric qualities of the WHOQOL-BREF(PT) (the questionnaire developed by the World Health Organization Quality of Life Grpup for quality of life assessment), when applied to Portuguese elderly people residing in a community setting. The psychometric qualities were assessed by confirmatory factor analysis. A hierarchical second-order model and a third model were performed, and all three models presented similar and reasonable adjustment indexes. The data analysis showed that the construct failed only regarding discriminant validity because the correlations between the first-order factors were higher, associated with lower values of average variance extracted. The psychometric qualities found in the original translation/validation of the WHOQOL-BREF(PT) were compared with those found in this study; this study found higher correlations between domains but a similar level of factor reliability. The findings of this study lead to three recommendations: (i) to compute each factor score for each participant using the factor score weights obtained from confirmatory analysis models instead of adopting a unitary weight for each item, as proposed by the authors of the original translation/validation of the WHOQOL-BREF(PT); (ii) to compute a QOL score, which is not included in the original translation/validation; and (iii) to analyze differences between individual scores for each participants, which should be done by a group of health experts.

Concentration of mercury species in hair, blood and urine of individuals occupationally exposed to gaseous elemental mercury in Asturias (Spain) and its comparison with individuals from a control group formed by close relatives.

Between November 19th, 2012 and December 3rd, 2012, 50 workers were intoxicated with gaseous Hg in San Juan de Nieva (Asturias, Spain) during the maintenance of a heat exchanger of a zinc manufacturer. We have quantified the concentration of methylmercury (MeHg), ethylmercury (EtHg) and Hg(II) in blood, hair and urine samples of those individuals taken three years after the accident. Blood, hair and urine of their closest relatives were also analyzed to assess whether the mercury burden present in the intoxicated individuals was due to the occupational exposure or to environmental or lifestyle-related factors. The determination of the mercury species in the samples was carried out applying multiple spiking Isotope Dilution GC-ICP-MS. This procedure corrects for possible interconversion reactions between the Hg species during the sample preparation procedure. Linear correlations were observed for both groups when plotting MeHg in blood vs MeHg in hair, and MeHg in hair vs Hg (II) in urine. The concentrations of Hg species in the intoxicated individuals were not significantly different from those obtained in the control group except for MeHg in blood. Significantly higher levels of MeHg in blood were obtained in some of the intoxicated individuals who had not consumed fish or seafood since the accident. A different correlation between MeHg in hair and MeHg in blood was obtained for these individuals compared to the control group who showed a hair-to-blood ratio consistent with the reported value for people exposed to Hg via fish consumption. Our results suggest that ingested MeHg followed the same pathway of deposition in hair in exposed and non-exposed individuals. However, the exposed individuals with high MeHg levels in blood showed a significantly different extent of MeHg deposition in hair compared to the control group.