Effectiveness of educational programs in hearing health: a systematic review and meta-analysis.

OBJECTIVE: To evaluate the effectiveness of hearing health education programs aimed at preventing noise-induced hearing loss (NIHL), of recreational and occupational origin, by means of a systematic review and meta-analysis. DESIGN: The search strategy was carried out in on five electronic databases, as well as referrals from experts. The risk of bias was judged, and the random effects meta-analysis was performed. The certainty of the evidence was assessed. STUDY SAMPLE: Effectiveness studies that used educational intervention in hearing health and prevention of NIHL were included. RESULTS: 42 studies were included. The Dangerous Decibels program was the only one that could be quantitatively analysed and showed improvement in the post-intervention period of up to one week [SMD = 0.60; CI95% = 0.38-0.82; I2 = 92.5%) and after eight weeks [SMD = 0.45; CI95% = 0.26-0.63; I2 = 81.6%) compared to the baseline. The certainty of evidence was judged as very low. CONCLUSIONS: The Dangerous Decibels program is effective after eight weeks of intervention. The other programs cannot be quantified. They still present uncertainty about their effectiveness. The level of certainty is still low for this assessment.

Copper drives remodeling of metabolic state and progression of clear cell renal cell carcinoma.

Copper (Cu) is an essential trace element required for mitochondrial respiration. Late-stage clear cell renal cell carcinoma (ccRCC) accumulates Cu and allocates it to mitochondrial cytochrome c oxidase. We show that Cu drives coordinated metabolic remodeling of bioenergy, biosynthesis and redox homeostasis, promoting tumor growth and progression of ccRCC. Specifically, Cu induces TCA cycle-dependent oxidation of glucose and its utilization for glutathione biosynthesis to protect against H 2 O 2 generated during mitochondrial respiration, therefore coordinating bioenergy production with redox protection. scRNA-seq determined that ccRCC progression involves increased expression of subunits of respiratory complexes, genes in glutathione and Cu metabolism, and NRF2 targets, alongside a decrease in HIF activity, a hallmark of ccRCC. Spatial transcriptomics identified that proliferating cancer cells are embedded in clusters of cells with oxidative metabolism supporting effects of metabolic states on ccRCC progression. Our work establishes novel vulnerabilities with potential for therapeutic interventions in ccRCC. Accumulation of copper is associated with progression and relapse of ccRCC and drives tumor growth.Cu accumulation and allocation to cytochrome c oxidase (CuCOX) remodels metabolism coupling energy production and nucleotide biosynthesis with maintenance of redox homeostasis.Cu induces oxidative phosphorylation via alterations in the mitochondrial proteome and lipidome necessary for the formation of the respiratory supercomplexes. Cu stimulates glutathione biosynthesis and glutathione derived specifically from glucose is necessary for survival of Cu Hi cells. Biosynthesis of glucose-derived glutathione requires activity of glutamyl pyruvate transaminase 2, entry of glucose-derived pyruvate to mitochondria via alanine, and the glutamate exporter, SLC25A22. Glutathione derived from glucose maintains redox homeostasis in Cu-treated cells, reducing Cu-H 2 O 2 Fenton-like reaction mediated cell death. Progression of human ccRCC is associated with gene expression signature characterized by induction of ETC/OxPhos/GSH/Cu-related genes and decrease in HIF/glycolytic genes in subpopulations of cancer cells. Enhanced, concordant expression of genes related to ETC/OxPhos, GSH, and Cu characterizes metabolically active subpopulations of ccRCC cells in regions adjacent to proliferative subpopulations of ccRCC cells, implicating oxidative metabolism in supporting tumor growth.

A miniaturized sample preparation method for routine elemental determination in whole blood using volumetric absorptive micro-sampling by ICP-QQQ.

Volumetric absorptive micro-sampling (VAMS) has emerged as a simple and safe tool for collecting and storing blood samples in clinical and bioanalytical fields. This study presents a novel method for determining essential and non-essential trace elements (As, Be, Cd, Cs, Cu, Fe, Mg, P, Pb, S, Sb, Se, Tl, V, U) in VAMS-collected blood samples using microwave-assisted digestion with diluted acid as sample preparation method and an inductively coupled plasma triple quadrupole mass spectrometry (ICP-QQQ) as determination technique. While certain elements posed challenges due to VAMS tip background issues (Al, Ti, Cr, Mn, Co, Ni, Sn, Mo, Ba), the method demonstrated high precision and accuracy for the targeted analytes. It was demonstrated that 4.5 mol L-1 HNO3 plus 100 µL H2O2 30% (w/w) was suitable for an efficiency of digestion for further elemental determination using micro-analysis (spending less than 300 µL analytical solution) by ICP-QQQ, given that the residual carbon content (RCC) after the digestion procedure was lower than 5%. All the results higher than limit of quantification (LOQ) were in agreement with reference values for all analytes. Accuracy was assessed through reference material analysis and recovery tests using spiked samples. Moreover, suitable agreements (p > 0.05) between this method (VAMS-M) and the comparative method (liquid sampling method) were obtained for all analytes >LOQ. Furthermore, all results >LOQ showed good precision according to precision requirements (Horwitz equation). In this way, with the use of dilute acid, low dilution factor (30-fold), and excellent digestion efficiency (>95%), the proposed method was able to achieve an excellent detection limit, precision, and accuracy for 15 elements: As, Be, Cd, Cs, Cu, Fe, Mg, P, Pb, S, Sb, Se, Tl, V, and U using ICP-MS/MS, without the need for matrix-matched calibration curves. This research showcases an innovative analytical approach using VAMS for blood samples, offering biosafety, practicality, sensitivity, versatility, and robustness. This method contributes to the advancement of trace element analysis in biomedical research and clinical applications.

Doppler Broadening of Neutron Cross-Sections Using Kaniadakis Entropy.

In the last seven years, Kaniadakis statistics, or κ-statistics, have been applied in reactor physics to obtain generalized nuclear data, which can encompass, for instance, situations that lie outside thermal equilibrium. In this sense, numerical and analytical solutions were developed for the Doppler broadening function using the κ-statistics. However, the accuracy and robustness of the developed solutions contemplating the κ distribution can only be appropriately verified if applied inside an official nuclear data processing code to calculate neutron cross-sections. Hence, the present work inserts an analytical solution for the deformed Doppler broadening cross-section inside the nuclear data processing code FRENDY, developed by the Japan Atomic Energy Agency. To do that, we applied a new computational method called the Faddeeva package, developed by MIT, to calculate error functions present in the analytical function. With this deformed solution inserted in the code, we were able to calculate, for the first time, deformed radiative capture cross-section data for four different nuclides. The usage of the Faddeeva package brought more accurate results when compared to other standard packages, reducing the percentage errors in the tail zone in relation to the numerical solution. The deformed cross-section data agreed with the expected behavior compared to the Maxwell-Boltzmann.

Influence of culinary treatment on the concentration and on the bioavailability of cadmium, chromium, copper, and lead in seafood.

BACKGROUND: Seafood present important advantages for human nutrition, but it can also accumulate high levels of toxic and potentially toxic elements. Culinary treatments could influence seafood chemical element content and element bioavailability. In this study, the influence of culinary treatments on the total concentration and on the bioavailability of Cd, Cr, Cu and Pb in shark, shrimp, squid, oyster, and scallop was assessed. METHODS: Boiling, frying, and sautéing with or without seasonings (salt, lemon juice and garlic) were evaluated. Total concentration and bioavailability of Cd, Cr, Cu and Pb in seafood after all these culinary treatments were compared with those in uncooked samples. Analytes were determined by triple-quadrupole inductively coupled plasma mass spectrometry (ICP-MS/MS). An alternative to express the results avoiding underestimated or overestimated values was proposed. RESULTS: The analytes concentration in seafood without culinary treatment varied from 0.0030 µg g-1 (shrimp) to 0.338 µg g-1 (oyster) for Cd; 0.010 µg g-1 (squid) to 0.036 µg g-1 (oyster) for Cr; 0.088 µg g-1 (scallop) to 8.63 µg g-1 (oyster) for Cu, and < 0.005 µg g-1 (shrimp, squid and oyster) to 0.020 µg g-1 (shark) for Pb. Only Cd (in scallop) was influenced by culinary treatments (reduction from 37 to 53 % after boiling, frying, and sautéing). Bioavailability percentage varied from 11% (oyster) for Cd; 18% (oyster) to 41% (shark) for Cr; 6% (shark) for Cu, and 8% (oyster) for Pb. Bioavailability percentage was not influenced by culinary treatments. CONCLUSION: Cadmium concentration was reduced in scallop after some culinary treatments (reduction o 37-53% after boiling, frying, and sautéing), but bioavailability percentage was not influenced. The employed analytical method was adequate for the purpose, presenting import results for food safety assessment about the influence of culinary treatments on metals concentration and bioavailability in seafood.

Single-molecule chemistry. Part I: monitoring oxidation of G in oligonucleotides using CY3 fluorescence.

Single-molecule hybridisation of CY3 dye labelled short oligonucleotides to surface immobilised probes was investigated in zero-mode waveguide nanostructures using a modified DNA sequencer. At longer measuring times, we observed changes of the initial hybridisation fluorescence pulse pattern which we attribute to products created by chemical reactions at the nucleobases. The origin is a charge separated state created by a photoinduced electron transfer from nucleobases to the dye followed by secondary reactions with oxygen and water, respectively. The positive charge can migrate through the hybrid resulting in base modifications at distant sites. Static fluorescence spectra were recorded in order to determine the properties of CY3 stacking to different base pairs, and compared to pulse intensities. A characteristic pulse pattern change was assigned to the oxidation of G to 8-oG besides the formation of a number of secondary products that are not yet identified. Further, we present a method to visualise the degree of chemical reactions to gain an overview of ongoing processes. Our study demonstrates that CY3 is able to oxidise nucleobases in ds DNA, and also in ss overhangs. An important finding is the correlation between nucleobase oxidation potential and fluorescence quenching which explains the intensity changes observed in single molecule measurements. The analysis of fluorescence traces provides the opportunity to track complete and coherent reaction sequences enabling to follow the fate of a single molecule over a long period of time, and to observe chemical reactions in real-time. This opens up the opportunity to analyse reaction pathways, to detect new products and short-lived intermediates, and to investigate rare events due to the large number of single molecules observed in parallel.

Hearing prevention program: how speech-language pathologists and audiologists work in the occupational health area / Programa de preservação auditiva: como atuam os fonoaudiólogos da área da saúde do trabalhador

ABSTRACT Objective: to characterize speech-language pathologists/audiologists and their interventions in the occupational health area, regarding the Hearing Prevention Program (HPP). Methods: a cross-sectional, quantitative study with 74 speech-language pathologists/audiologists from several regions of Brazil, working in the areas of Occupational Health and Audiology. The participants answered a questionnaire on their professional and educational profile, as well as on their work related to the HCP components. For data analysis, statistical procedures were used (Chi-Square Test and the test of difference in proportions, at the significance level of 0.05 - 5%), in relation to the variables of gender, age, job position, time since their graduation and academic training in occupational safety and health. Results: the speech-language pathologists/audiologists, mostly women, who reported performing the HPP, were older than 30 years, had specific postgraduate courses, but still had doubts about the Program implementation, they worked for companies with over 1,000 employees, disregarding their employment status, performing interventions, such as: hearing management and guidance on the correct use of ear protectors. Conclusion: a high number of speech-language pathologists in the Occupational Health area have not developed a complete hearing conservation program to this date.

RESUMO Objetivo: caracterizar os fonoaudiólogos e as ações que desenvolvem na área da saúde do trabalhador com relação ao Programa de Preservação Auditiva (PPA). Métodos: estudo quantitativo, transversal, com 74 fonoaudiólogos de diversas regiões do Brasil que trabalham em Saúde do Trabalhador e Audiologia. Os participantes responderam a um questionário sobre seu perfil profissional, formação e atuação em relação aos componentes do PPA. Para a análise dos dados utilizou-se procedimentos estatísticos (teste Qui-Quadrado e o teste de diferença de proporções, ao nível de significância 0,05 - 5%) em relação às variáveis sexo, idade, função, tempo de formado e formação acadêmica em saúde e segurança do trabalho. Resultados: os fonoaudiólogos, a maioria mulheres, que relataram realizar o PPA, tinham idade superior a 30 anos; fizeram cursos de pós-graduação específica, mas ainda apresentavam dúvidas em relação à implantação do Programa; estavam em empresas com mais de 1000 funcionários, independentemente de seu regime de trabalho, realizando ações como: gerenciamento auditivo e orientação sobre utilização correta de protetores auriculares. Conclusão: há ainda um número elevado de fonoaudiólogos na área de Saúde do Trabalhador que não desenvolvem um programa de preservação auditiva na íntegra.

Maxwell-Wagner Effect Applied to Microwave-Induced Self-Ignition: A Novel Approach for Carbon-Based Materials.

A new method for analytical applications based on the Maxwell-Wagner effect is proposed. Considering the interaction of carbonaceous materials with an electromagnetic field in the microwave frequency range, a very fast heating is observed due to interfacial polarization that results in localized microplasma formation. Such effect was evaluated in this work using a monomode microwave system, and temperature was recorded using an infrared camera. For analytical applications, a closed reactor under oxygen pressure was evaluated. The combination of high temperature and oxidant atmosphere resulted in a very effective self-ignition reaction of sample, allowing its use as sample preparation procedure for further elemental analysis. After optimization, a high sample mass (up to 600 mg of coal and graphite) was efficiently digested using only 4 mol L-1 HNO3 as absorbing solution. Several elements (Ba, Ca, Fe, K, Li, Mg, Na, and Zn) were determined by inductively coupled plasma optical emission spectrometry (ICP-OES). Accuracy was evaluated by using a certified reference material (NIST 1632b). Blanks were negligible, and only a diluted solution was required for analytes absorption preventing residue generation and making the proposed method in agreement with green chemistry recommendations. The feasibility of the proposed method for hard-to-digest materials, the minimization of reagent consumption, and the possibility of multi elemental analysis with lower blanks and better limits of detection can be considered as the main advantages of this method.

A feasible method for As speciation in several types of seafood by LC-ICP-MS/MS.

A method for arsenic speciation in shark, shrimp, squid, oyster and scallop using liquid chromatography coupled to inductively coupled plasma triple quadrupole mass spectrometry (LC-ICP-MS/MS) was proposed. Suitable sensitivity and selectivity by LC-ICP-MS/MS were obtained using 10 mmol L-1 (NH4)2HPO4 diluted in 1% methanol (pH 8.65) as mobile phase. Recoveries from 90 to 104% for arsenobetaine (AsB), arsenite [As(III)], dimethylarsinic acid (DMA), monomethylarsonic acid (MMA) and arsenate [As(V)] were obtained for all samples. A certificated reference material was also analyzed and the sum of As species was in agreement with the total As concentration. Limits of quantification (LOQ) for AsB, As(III), DMA, MMA, and As(V) were 6, 30, 6, 12 and 26 ng g-1, respectively. Higher concentration of AsB was found in all seafood, while As(III) and DMA were found only in oyster. Arsenate was found in squid and scallops, and MMA was below the LOQ in all samples.

Dynamic Myofibrillar Remodeling in Live Cardiomyocytes under Static Stretch.

An increase in mechanical load in the heart causes cardiac hypertrophy, either physiologically (heart development, exercise and pregnancy) or pathologically (high blood pressure and heart-valve regurgitation). Understanding cardiac hypertrophy is critical to comprehending the mechanisms of heart development and treatment of heart disease. However, the major molecular event that occurs during physiological or pathological hypertrophy is the dynamic process of sarcomeric addition, and it has not been observed. In this study, a custom-built second harmonic generation (SHG) confocal microscope was used to study dynamic sarcomeric addition in single neonatal CMs in a 3D culture system under acute, uniaxial, static, sustained stretch. Here we report, for the first time, live-cell observations of various modes of dynamic sarcomeric addition (and how these real-time images compare to static images from hypertrophic hearts reported in the literature): 1) Insertion in the mid-region or addition at the end of a myofibril; 2) Sequential addition with an existing myofibril as a template; and 3) Longitudinal splitting of an existing myofibril. The 3D cell culture system developed on a deformable substrate affixed to a stretcher and the SHG live-cell imaging technique are unique tools for real-time analysis of cultured models of hypertrophy.

Microfluidics-based laser cell-micropatterning system.

The ability to place individual cells into an engineered microenvironment in a cell-culture model is critical for the study of in vivo relevant cell-cell and cell-extracellular matrix interactions. Microfluidics provides a high-throughput modality to inject various cell types into a microenvironment. Laser guided systems provide the high spatial and temporal resolution necessary for single-cell micropatterning. Combining these two techniques, the authors designed, constructed, tested and evaluated (1) a novel removable microfluidics-based cell-delivery biochip and (2) a combined system that uses the novel biochip coupled with a laser guided cell-micropatterning system to place individual cells into both two-dimensional (2D) and three-dimensional (3D) arrays. Cell-suspensions of chick forebrain neurons and glial cells were loaded into their respective inlet reservoirs and traversed the microfluidic channels until reaching the outlet ports. Individual cells were trapped and guided from the outlet of a microfluidic channel to a target site on the cell-culture substrate. At the target site, 2D and 3D pattern arrays were constructed with micron-level accuracy. Single-cell manipulation was accomplished at a rate of 150 µm s(-1) in the radial plane and 50 µm s(-1) in the axial direction of the laser beam. Results demonstrated that a single-cell can typically be patterned in 20-30 s, and that highly accurate and reproducible cellular arrays and systems can be achieved through coupling the microfluidics-based cell-delivery biochip with the laser guided system.

Laser cell-micropatterned pair of cardiomyocytes: the relationship between basement membrane development and gap junction maturation.

The basement membrane (BM), a network of laminin and collagen IV, mechanically supports individual cells and directly mediates cell-cell and cell-extracellular matrix (ECM) interactions. For example, the BM network that tightly encloses each cardiomyocyte (CM) mediates the alignment of CMs with collagen I in the ECM. Additionally, the BM-laminin is involved in the formation of gap junctions (GJs), which regulate electrical coupling between two CMs in the myocardium. The role of BM in GJ maturation remains unclear because of the complicated in vivo structures and lack of an ideal in vitro culturing mode. In this study, our laser cell-micropatterning system was used to place two neonatal CMs (NCMs) in contact on an aligned collagen gel (ACG) to study the relationship between GJ maturation and BM development. The results of double immunofluorescence staining and confocal imaging showed that BM-laminin was deposited earlier than the formation of GJs in the intercellular space and that newly expressed connexin 43 clusters were preferentially assembled near the deposited BM structures. Eventually the BM network surrounded the GJs.

Effect of simultaneous cooling on microwave-assisted wet digestion of biological samples with diluted nitric acid and O2 pressure.

The present work evaluates the influence of vessel cooling simultaneously to microwave-assisted digestion performed in a closed system with diluted HNO3 under O2 pressure. The effect of outside air flow-rates (60-190 m(3) h(-1)) used for cooling of digestion vessels was evaluated. An improvement in digestion efficiency caused by the reduction of HNO3 partial pressure was observed when using higher air flow-rate (190 m(3) h(-1)), decreasing the residual carbon content for whole milk powder from 21.7 to 9.3% (lowest and highest air flow-rate, respectively). The use of high air flow-rate outside the digestion vessel resulted in a higher temperature gradient between liquid and gas phases inside the digestion vessel and improved the efficiency of sample digestion. Since a more pronounced temperature gradient was obtained, it contributed for increasing the condensation rate and thus allowed a reduction in the HNO3 partial pressure of the digestion vessel, which improved the regeneration of HNO3. An air flow-rate of 190 m(3) h(-1) was selected for digestion of animal fat, bovine liver, ground soybean, non fat milk powder, oregano leaves, potato starch and whole milk powder samples, and a standard reference material of apple leaves (NIST 1515), bovine liver (NIST 1577) and whole milk powder (NIST 8435) for further metals determination by inductively coupled plasma atomic emission spectroscopy (ICP-OES). Results were in agreement with certified values and no interferences caused by matrix effects during the determination step were observed.

Desorption electrospray ionization mass spectrometry (DESI-MS) applied to the speciation of arsenic compounds from fern leaves.

The different chemical forms of arsenic compounds, including inorganic and organic species, present distinct environmental impacts and toxicities. Desorption electrospray ionization mass spectrometry (DESI-MS) is an excellent technique for in situ analysis, as it operates under atmospheric pressure and room temperature and is conducted with no/minimal sample pretreatment. Aimed at expanding its scope, DESI-MS is applied herein for the quick and reliable detection of inorganic (arsenate--As(V): AsO4(3-) and arsenite--As(III): AsO2(-)) and organic (dimethylarsinic acid--DMA: (CH3)2AsO(OH) and disodium methyl arsonate hexahydrate: CH3AsO3·2Na·6H2O) arsenic compounds in fern leaves. Operational conditions of DESI-MS were optimized with DMA standard deposited on paper surfaces to improve ionization efficiency and detection limits. Mass spectra data for all arsenic species were acquired in both the positive and negative ion modes. The positive ion mode was shown to be useful in detecting both the organic and inorganic arsenic compounds. The negative ion mode was shown only to be useful in detecting As(V) species. Moreover, MS/MS spectra were recorded to confirm the identity of each arsenic compound by the characteristic fragmentation profiles. Optimized conditions of DESI-MS were applied to the analysis of fern leaves. LC-ICP-MS was employed to confirm the results obtained by DESI-MS and to quantify the arsenic species in fern leaves. The results confirmed the applicability of DESI-MS in detecting arsenic compounds in complex matrices.

Laser guidance-based cell detection in a microfluidic biochip.

We developed a microfluidic biochip to perform laser guidance on two cell types, chick embryonic forebrain neurons and spinal cord neurons. Observation of neurons under a high-magnification microscope, which we obtained from these two cell types, showed no difference in morphology. However, when flowing in the microfluidic channel and simultaneously being laser guided, the two cell types gained quite different guidance speeds under the same experimental conditions. The results demonstrate that different cell types with the same morphology (e.g., size, shape, etc.) can be effectively distinguished from each other by measuring the difference in guidance speeds (the maximum flow speed minus the initial flow speed). This technique is expected to provide a new approach to high-throughput, label-free cell sorting with high sensitivity.

Evaluation of central auditory discrimination abilities in older adults.

The present study focuses on auditory discrimination abilities in older adults aged 65-89 years. We applied the "Leipzig inventory for patient psychoacoustic" (LIPP), a psychoacoustic test battery specifically designed to identify deficits in central auditory processing. These tests quantify the just noticeable differences (JND) for the three basic acoustic parameters (i.e., frequency, intensity, and signal duration). Three different test modes [monaural, dichotic signal/noise (s/n) and interaural] were used, stimulus level was 35 dB sensation level. The tests are designed as three-alternative forced-choice procedure with a maximum-likelihood procedure estimating p = 0.5 correct response value. These procedures have proven to be highly efficient and provide a reliable outcome. The measurements yielded significant age-dependent deteriorations in the ability to discriminate single acoustic features pointing to progressive impairments in central auditory processing. The degree of deterioration was correlated to the different acoustic features and to the test modes. Most prominent, interaural frequency and signal duration discrimination at low test frequencies was elevated which indicates a deterioration of time- and phase-dependent processing at brain stem and cortical levels. LIPP proves to be an effective tool to identify basic pathophysiological mechanisms and the source of a specific impairment in auditory processing of the elderly.

Evaluation of oxygen pressurized microwave-assisted digestion of botanical materials using diluted nitric acid.

The feasibility of diluted nitric acid solutions for microwave-assisted decomposition of botanical samples in closed vessels was evaluated. Oxygen pressurized atmosphere was used to improve the digestion efficiency and Al, Ca, K, Fe, Mg and Na were determined in digests by inductively coupled plasma optical emission spectrometry (ICP OES). Efficiency of digestion was evaluated taking into account the residual carbon content (RCC) and residual acidity in digests. Samples were digested using nitric acid solutions (2, 3, 7, and 14 mol L(-1) HNO(3)) and the effect of gas phase composition inside the reaction vessels by purging the vessel with Ar (inert atmosphere, 1 bar), air (20% of oxygen, 1 bar) and pure O(2) (100% of oxygen, 1 bar) was evaluated. The influence of oxygen pressure was studied using pressures of 5, 10, 15 and 20 bar. It was demonstrated that a diluted nitric acid solution as low as 3 mol L(-1) was suitable for an efficient digestion of sample masses up to 500 mg of botanical samples using 5 bar of oxygen pressure. The residual acidities in final digests were lower than 45% in relation to the initial amount of acid used for digestion (equivalent to 1.3 mol L(-1) HNO(3)). The accuracy of the proposed procedure was evaluated using certified reference materials of olive leaves, apple leaves, peach leaves and pine needles. Using the optimized conditions for sample digestion, the results obtained were in agreement with certified values. The limit of quantification was improved up to a factor of 14.5 times for the analytes evaluated. In addition, the proposed procedure was in agreement with the recommendations of the green chemistry once it was possible to obtain relatively high digestion efficiency (RCC<5%) using only diluted HNO(3), which is important to minimize the generation of laboratory residues.